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Grant Number: 1C06RR016518-01 PI Name: KOHLER, PETER O. PI Title: PRESIDENT Project Title: EXTRAMURAL RES FACILITIES CONSTR PROJECTS Abstract: This abstract is not available. Thesaurus Terms: There are no thesaurus terms on file for this project. Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098
Fiscal Year: 2001
Grant Number: 1C06RR017585-01 PI Name: KOHLER, PETER O. PI Email: PI Title: PRESIDENT Project Title: EXTRAMURAL RESEARCH FACILITIES CONSTRUCTION Abstract: DESCRIPTION (provided by applicant): This application requests funds to construct Phase II infill to the Animal Services Building (ASB) addition that will provide housing for 544 non-human primates (NHPs) in an 11,970 sq. ft. state-of-the art facility. The expansion will enable the Oregon Regional Primate Research Center (ORPRC) to meet rapidly growing research needs, as well as comply with new United States Department of Agriculture (USDA) mandated standards for expanded social housing. The Oregon Health and Science University (OHSU) made a major commitment by providing approximately $4 million in funds to construct a 35,600 sq. ft. shell addition to the ASB. Phase I infill consists of a 13,000 sq. ft. facility that will house up to 192 NHPs, as well as provide behavioral testing rooms, and office and new cage wash areas. Rapid growth in various research areas necessitates additional NHP housing. Current NHP housing is full, resulting in the following problems: 1) limited ability to meet the needs of existing research programs or begin new projects; 2) delayed procurement of needed animals; 3) delayed renovations due to lack of flex space; and 4) delayed implementation of social housing. The Specific Aims of this application are to construct Phase II infill of the ASB addition, consisting of Animal Biosafety Level 2 (ABL2) containment housing for up to 544 NHPs in 17 animal holding rooms, a feed storage room and a procedure room. Each animal holding room will contain 32 NHPs in paired cages, as well as large group play enclosures to permit groups of animals to regularly experience an enhanced social environment. Thesaurus Terms: There are no thesaurus terms on file for this project. Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2002 Department: NONE Project Start: 15-SEP-2002 Project End: 14-SEP-2004 ICD: NATIONAL CENTER FOR RESEARCH RESOURCES IRG: STRB
Grant Number: 1G20RR016033-01 PI Name: KOHLER, PETER O. PI Email: PI Title: PRESIDENT Project Title: PRIMARY HOUSING UNITS FOR NON-HUMAN PRIMATES Abstract: This abstract is not available. Thesaurus Terms: There are no thesaurus terms on file for this project. Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2000 Department: NONE Project Start: 30-SEP-2000 Project End: 29-SEP-2004 ICD: NATIONAL CENTER FOR RESEARCH RESOURCES IRG: STRB
Grant Number: 1G20RR016823-01 PI Name: KOHLER, PETER O. PI Email: PI Title: PRESIDENT Project Title: SMALL GROUP HOUSING FOR AIDS/SPF MONKEYS Abstract: This abstract is not available. Thesaurus Terms: There are no thesaurus terms on file for this project. Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2001 Department: MEDICINE Project Start: 01-SEP-2001 Project End: 31-AUG-2004 ICD: NATIONAL CENTER FOR RESEARCH RESOURCES IRG: STRB
Grant Number: 1G20RR018326-01 PI Name: SMITH, M SUSAN. PI Email: smithsu@ohsu.edu PI Title: ASSOCIATE PROFESSOR OF PHYSIOLOGY Project Title: RHESUS CORRAL UPGRADE Abstract: DESCRIPTION (provided by applicant): The renovation project is part of a long-range plan to expand rhesus macaque (Macaca mulatta) production at the ONPRC. The specific aim of this application is to alter and renovate the 30-year-old facilities associated with six one-acre corrals that hold SPF Indian rhesus macaque breeding colonies. The continued use of the corrals for rhesus macaque production is driven by the following: 1) the growth in research projects and the numbers of NHPs assigned to projects requires a substantial increase in the number of available NHPs; 2) the ONPRC is a national resource and must provide accessibility to investigators outside of the Center; 3) the long-range plan calls for the doubling of SPF breeding program to create a self-sustaining supply of NHPs; 4) the reserve capacity for animal housing must be expanded for proper animal management; and 5) the corrals are a cost effective method for breeding and raising animals into adulthood in a complex and environmentally rich social structure. Animals in the corrals are used as juveniles (research programs in AIDS, infectious diseases, developmental neurobiology), adults (research programs in immunology, neuroscience and reproduction) and retired breeders (research programs in aging). The renovation of the corral facilities will make them suitable for long-term use and includes the following: 1) convert the storage area between corrals four and five into a sheltered feeding area for corral five; 2) convert the existing corral five feeding area into an area for safe handling of animals; 3) upgrade the infrastructure (electric service, domestic water, sanitary sewer) for all rhesus corral facilities; 4) replace the deteriorated roofs on all feeding and holding areas; 5) replace damaged and worn metal panels on all structures; 6) repair and reseal concrete in all feeding and holding areas; 7) install additional radiant heaters and lights in all feeding and holding areas; 8) install a water pressure booster pump for wash down systems; 9) add perching to all feeding areas; and 10) replace deteriorating animal enrichment equipment in all corrals. This application will provide funds for the renovation of facilities associated with six one-acre corrals. The renovation is necessary for continued use of the corrals to expand rhesus macaque production and to comply with United States Department of Agriculture (USDA) policies and provisions of the Animal Welfare Act (AWA). Thesaurus Terms: Macaca mulatta, animal colony, building /facility design /renovation animal breeding, animal care
Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-FEB-2003 Project End: 31-JAN-2004 ICD: NATIONAL CENTER FOR RESEARCH RESOURCES IRG: STRB
Grant Number: 1G20RR018457-01 PI Name: SMITH, M SUSAN. PI Email: smithsu@ohsu.edu PI Title: ASSOCIATE PROFESSOR Project Title: AIDS RELATED RESEARCH CAGING Abstract: DESCRIPTION (provided by applicant): The ONPRC proposes to purchase caging suitable for housing SPF male Indian-origin rhesus macaques (Macaca mulatta) used in AIDS related research. Increasing demands for SPF rhesus macaques for AIDS related research at the ONPRC and nationally has exceeded the supply efforts to expand production colonies to supply more animals has necessitated retention of female offspring for breeding, leaving male SPF rhesus macaques as the primary model for AIDS-related research projects. The Animal Biosafety Level 2 and 3 (ABLS2 and ABSL3) containment facility which houses animals for AIDS-related research at the ONPRC is not equipped with cages suitable for Group 4 male rhesus macaques. The existing cages do not have the 6.0 sq. ft. of floor space required by USDA and do not provide the option of social housing. It is very important that animals used in AIDS-related research have the opportunity to experience an enriched social environment. Requested in this application are 6.2 sq. ft. cages and associated equipment that will comply with USDA requirements for Group 4 male rhesus macaques and provide a wide range of options for environmental enrichment and social housing of NHPs used in AIDS-related research. Thesaurus Terms: AIDS, Macaca mulatta, animal care, animal colony, biomedical equipment purchase Institution:
OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-FEB-2003 Project End: 31-JAN-2004 ICD: NATIONAL CENTER FOR RESEARCH RESOURCES IRG: STRB
Grant Number: 1R01DK060685-01A2 PI Name: GROVE, KEVIN L. PI Email: grovek@ohsu.edu PI Title: Project Title: NPY Feeding Circuits during Development Abstract: Obesity is now considered a worldwide health concern, being a major contributor to the increased incidences of coronary heart disease and type II diabetes. By 1998, 18% of the adults in the United states were defined as being obese, with greater than twice that number categorized as overweight. Furthermore, the number of obese adults in the United States is increasing at a rate of 0.7% per year. Even more disturbing is the dramatic increase in obesity and type II diabetes among children. The central hypothesis of this proposal is that body weight management during adulthood is directly determined by the hypothalamic feeding circuits established during a "critical period" postnatal development. Furthermore, if exposure to perturbations in energy balance occurs during this "critical period" of neural plasticity, permanent alterations in body weight management may occur and lead to abnormal body weight phenotypes during adulthood. One of the most potent modulators of appetite and energy expenditure in the hypothalamus is neuropeptide Y (NPY). There are dynamic changes in the hypothalamic NPY system during postnatal development that implicate this system as being pivotal for the proper maturation of hypothalamic feeding circuitry. This proposal will study the postnatal period to 1) Determine the functional importance of the development of ARH projections. 2) Determine if NPY plays a role in the regulation of body weight management during early postnatal development. 3) Determine if changes in the endogenous NPY system are responsible for the obese phenotype induced by chronic overfeeding during the postnatal period. The main goal of this proposal is to use a multidisciplinary approach to determine if modification of the endogenous NPY system during postnatal development leads to abnormal body weight management during adulthood. To identify the role of the endogenous NPY system in the regulation of food intake and energy expenditure during the postnatal period we will use a combination of in vivo (changes in food intake and adiposity in whole animals) and in vitro (peptide release and electrophysiology in hypothalamic explants) physiological and pharmacological experiments, coupled with neuroanatomical measures (immunocytochemistry and in situ hybridization). The changes in the hypothalamic NPY system in these models will be correlated with changes in peripheral markers of energy expenditure and body weight status, using RIA and real-time PCR. These studies will provide important insight into the consequences of manipulation of the NPY feeding circuits, during the postnatal period on metabolic rate and body weight during childhood. Understanding of the normal and abnormal development of this circuitry is critical to determining the physiological mechanisms that underlie adult obesity and identify a critical period for possible intervention. Thesaurus Terms: appetite regulatory center, bioenergetics, body weight, developmental neurobiology, hypothalamus, neuropeptide Y, weight control appetite, dietary excess, electrophysiology, glutamate, melanocyte stimulating hormone, neural plasticity, nutrient intake activity, obesity, phenotype immunocytochemistry, in situ hybridization, laboratory rat, nutrition related tag, polymerase chain reaction, radioimmunoassay
Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 05-MAY-2003 Project End: 30-APR-2008 ICD: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES IRG: END
Grant Number: 1R01DK065900-01 PI Name: SIMERLY, RICHARD B. PI Email: simerlyr@ohsu.edu PI Title: SINIOR SCIENTIST Project Title: Development of Leptin-Sensitive Hypothalamic Pathways Abstract: DESCRIPTION (provided by applicant): Neonatal factors that contribute to obesity are poorly understood. The long-range goal of this research is to clarify how the adipocyte-derived hormone leptin influences development of neuroendocrine pathways that regulate mammalian energy homeostasis. Central to this goal is determining how leptin affects development of neural projections from the arcuate nucleus of the hypothalamus (ARH), a key site for integration of information related to peripheral energy stores. Leptin signals are conveyed via the ARH to other brain regions involved in the regulation of body weight, such as the paraventricular nucleus of the hypothalamus (PVH), an important component of the final common pathway for regulation of energy metabolism. Evidence presented in the body of this proposal supports the concept that leptin functions as a developmental factor during neonatal life by directing formation of neural circuits involved in the control of feeding and energy balance. The overall hypothesis addressed in this proposal is that leptin acts directly on the ARH during a restricted neonatal critical period to promote formation of neural projections to the PVH involved in the regulation of body weight. We propose that the postnatal leptin surge has an enduring effect on ARH projections, and that developmental perturbations in leptin signaling cause permanent changes in neural pathways that transmit leptin signals in mature animals. Both physiological and in vitro experimental approaches will be used to test this hypothesis by addressing the following specific aims. Specific Aim 1. We propose to use axonal labeling methods and histochemical techniques to define the organization and development of ARH projections in obese mice that lack leptin (ob/ob mice) and in diabetic mice that lack a functional long form of its receptor (db/db mice). Specific Aim 2. We will also test the developmental activity of leptin by examining development of the ARH-PVH pathway in ob/ob and db/db mice treated with exogenous leptin, and determine if this developmental activity is restricted to a neonatal critical period. Specific Aim 3. To determine the site of action for leptin in directing development of the ARH-PVH pathway, we will utilize a new explant coculture assay, in addition to examining the direct effects of leptin on axon outgrowth from isolated ARH explants in vitro. Specific Aim 4. In addition, we will examine how nutritional manipulation of leptin levels impacts development of the ARH-PVH pathway. Specific Aim 5. Finally, we will determine if altered patterns of leptin dependent intrahypothalamic signaling accompany observed developmental changes in the ARH-PVH pathway. The results of the proposed research will expand our appreciation of leptin to include a profound developmental activity that promotes formation of leptin-responsive hypothalamic neural pathways. These studies may also provide essential clues about how the neonatal nutritional environment imposes enduring consequences on central regulation of feeding and energy balance throughout life. Thesaurus Terms: bioenergetics, biological signal transduction, developmental neurobiology, hypothalamus, leptin, neuroregulation, paraventricular nucleus, perinatal age difference, developmental nutrition, intercellular connection, neuroendocrine system, neuronal guidance, obesity fluorescent dye /probe, histochemistry /cytochemistry, laboratory mouse, newborn animal, nutrition related tag, radiotracer, transgenic animal Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 30-SEP-2003 Project End: 31-JUL-2006 ICD: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES IRG: END
Grant Number: 1R01MH065438-01A1 PI Name: OJEDA, SERGIO R. PI Email: ojedas@ohsu.edu PI Title: SCIENTIST/DIVISION HEAD Project Title: Molecular Specifiers of Neural Cell Plasticity Abstract: DESCRIPTION (provided by applicant): The initiation of mammalian puberty requires an increased pulsatile secretion of luteinizing hormone-releasing hormone (LHRH), the neuropeptide controlling sexual development, from specialized neurons in the basal hypothalamus. This increase is brought about by changes in the activity of neuronal and astroglial subsets functionally connected to the LHRH neuronal network. Whether such alterations in cell-cell communication are purely biochemical or require plastic rearrangements is not known. This application proposes a combination of cellular, molecular, and genomic approaches to examine the hypothesis that major changes in hypothalamic expression of three novel multigene families of cell-surface molecules recently implicated as critical components of synaptic specification and neuron-glia communication in the central nervous system, underlie the initiation of primate puberty. The demonstration of such changes will provide prima facie evidence for the concept that plastic rearrangements in neuron-to-neuron and glial-neuronal communication are integral components of the neuroendocrine mechanism by which the brain controls the onset of mammalian puberty. To this end, the following specific aims are proposed: 1. To test the hypothesis that coordinated changes in the hypothalamic expression of neurexins -- a recently discovered, multi-product family of cell-surface proteins thought to play a major role in neuronal cell recognition -- occur at the time of primate puberty. 2. To test the hypothesis that hypothalamic-specific changes in the expression of particular members of the novel protocadherin - a family of adhesion-signaling molecules recently implicated in the specification of brain synaptic connectivity, occur at the initiation of puberty. 3. To examine the hypothesis that hypothalamic-specific changes in the expression of a newly discovered family of neurexin-related molecules termed CASPRs (contactin-associated proteins) involved in glial-neuronal communication, and their interacting molecules, occurs at the time of primate puberty, and 4. To test the hypothesis that contactin/CASPR-1 and its astroglial receptor protein tyrosine phosphatase-beta (RPTP-b) mediate adhesive communication between astroglial cells and LHRH neurons, and that this is an event regulated by growth factors involved in the facilitatory control that glial cells exert on LHRH release during female sexual maturation. Thesaurus Terms: animal puberty, cell communication molecule, cell membrane, membrane protein, neural plasticity, neuron cadherin, cell adhesion molecule, glia, hypothalamus, protein tyrosine phosphatase Macaca mulatta, immunocytochemistry, laboratory mouse, laboratory rat, newborn animal, tissue /cell culture Institution:
OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-DEC-2002 Project End: 30-NOV-2006 ICD: NATIONAL INSTITUTE OF MENTAL HEALTH IRG: BCE
Grant Number: 1R13RR018799-01 PI Name: WOLF, DON P. PI Email: wolfd@ohsu.edu PI Title: SENIOR SCIENTIST Project Title: The ARTs in Action in Nonhuman Primates Abstract: DESCRIPTION (provided by applicant): The nonhuman primate (NHP) is a valuable model for studying human health with direct implications to biomedical investigations of infertility and contraception, infectious diseases and vaccine development, drug and alcohol addiction, brain disorders, gene and stem cell based therapy and transplant biology. There are significant needs for populations of specific pathogen-free (SPF) animals with unique genotypes that cannot be satisfied by the importation of animals from the wild or by the identification and propagation of founder animals by selective breeding. SPF, Indian-origin, rhesus macaques carrying specific MHC alleles is but one example. The (ARTs) have been practiced extensively in humans over the last 20 years and their applications in NHPs, while potentially very useful, have yet to be realized on a scale adequate to impact the needs described above. This application seeks support for a workshop that will identify and discuss integrative approaches to exploit the ARTs in NHPs thereby maximizing their contribution to ongoing and future research efforts. The overall topic of the workshop is based on the concept that research using NHPs can be improved significantly both in quality and quantity by increased availability of animals of desired genotypes, of genetically identical animals and of SPF animals. The aims of the workshop are to: 1) Review the current state of reproductive research employing the ARTs in NHPs including both basic concepts and specific applications; 2) Enhance interactions and collaborative efforts within and between NPRCs and academic institutions concerning reproductive research including infertility and contraceptive development studies that are clinically important but can't be accomplished in humans; and 3) Identify strategies that effectively expand NPRCs abilities to develop and practice the ARTs in NHPs. The proposed workshop will be held as a satellite meeting immediately prior to the annual meeting of the International Embryo Transfer Society. This arrangement will permit attendees to attend both meetings and should encourage participation by research trainees and fellows. Thesaurus Terms: Primate, assistive reproductive technique, biological model, meeting /conference /symposium contraceptive, fertility, genotype, interdisciplinary collaboration travel Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 30-SEP-2003 Project End: 29-SEP-2004 ICD: NATIONAL CENTER FOR RESEARCH RESOURCES IRG: ZRR1
Grant Number: 1R21MH063137-01A1 PI Name: MACHIDA, CURTIS A. PI Email: machidac@ohsu.edu PI Title: Project Title: Adrenergic Receptor Mechanisms in Antidepressant Therapy Abstract: Adrenergic receptors (ARs) serve as important regulators of central nervous system- (CNS-) mediated behavior and several neural functions, including mood, memory, neuroendocrine control, and stimulation of autonomic function. Alterations in adrenergic receptor number have been implicated in the pathophysiology of affective psychiatric disorders, including depression. Beta-adrenergic receptor (beta-AR) down-regulation occurs during chronic treatment with antidepressants, suggesting that the dysregulation of the betal-adrenerqic receptor (beta1-AR) subtype, the predominant beta- AR subtype in the brain, may be associated with depressive illness. We propose to develop a mechanistic understanding of transcriptional and post-transcriptional beta1-AR mRNA control during antidepressant therapy. We have examined the molecular mechanisms underlying beta1-AR mRNA down-regulation following agonist induction, and have identified potential transcriptional and post-transcriptional control mechanisms. Firstly, we have recently determined that the RNA binding factors HuR, hnRNP A1, and AUF-1 all interact with the 3' untranslated region (UTR) of the rat 61- AR mRNAs, and that HuR becomes induced in the presence of beta-AR agonist isoproterenol, resulting in the acceleration of _I-AR transcript degradation. Secondly, we have determined that exposure of C6 cells to isoproterenol results in a rapid induction of inducible cyclic AMP early repressor (ICER) and other related CREM (cyclic AMP response element (CRE) modulator) mRNA within two hours of stimulation, and serves to repress beta1-AR gene transcription. And thirdly, we have identified another transcriptional repressor region in the beta1-AR gene, encompassing positions -396 to -367, and have identified the repressor molecule as a novel bZlP-like transcription factor. We will chronically-infuse various antidepressants into rats and rhesus macaques, and recover cortical specimens to identify the specific molecular mechanisms underlying beta1-AR mRNA down-regulation. This information may provide insiahts in the molecular role of the adrenerqic receptors in depression, and develop a better understandinq of the efficacy of antidepressant treatment. Thus, this R21 grant submission is responsive to objective 4 of PA-00-073, "initial research and development for building significant future research". The primary experimental objectives of Specific Aim 1 are to verify that the RNA binding proteins HuR, hnRNP A1, and/or AUF-1 are the degradative molecules involved in antidepressant-induced beta1-AR mRNA down-regulation, and to determine whether antidepressants trigger the nucleocytoplasmic export of beta1-AR mRNAs, via interaction with HuR and other selective HuR ligands. The primary experimental objectives of Specific Aim 2 are to validate ICER and the novel bZlP-like transcription factor as potential repressors of beta1-AR gene expression during antidepressant therapy. Thesaurus Terms: beta adrenergic receptor, genetic regulation, pharmacokinetics, psychopharmacology RNA binding protein, antidepressant, clinical depression, gene expression, imipramine, isoproterenol, mental disorder chemotherapy, messenger RNA, receptor expression, transcription factor Macaca mulatta, laboratory rat Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-DEC-2002 Project End: 30-NOV-2004 ICD: NATIONAL INSTITUTE OF MENTAL HEALTH IRG: ZRG1
Grant Number: 3R01DK060685-01A2S1 PI Name: GROVE, KEVIN L. PI Email: grovek@ohsu.edu PI Title: Project Title: NPY Feeding Circuits during Development Abstract: This abstract is not available. Thesaurus Terms: There are no thesaurus terms on file for this project. Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 05-MAY-2003 Project End: 30-APR-2008 ICD: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES IRG: END
Grant Number: 5D43TW000668-09 PI Name: CONN, PAUL M. PI Email: PI Title: PROFESSOR & CHAIRMAN OF PHARMACOLOGY Project Title: INTERNATIONAL TRAINING AND RES. IN POPULATION AND HEALTH Abstract: This research training program is designed to enable NIH grant recipients of the Oregon Health Sciences University (OHSU) to continue to extend the geographic base of research and training efforts to Mexico and Chile, countries in which the dynamics of population growth negatively impacts public health, the environment and economic progress. This program includes pre-doctoral and sabbatical training opportunities in reproductive sciences. Because of the proximity of Mexico to the United States and the substantial length of the common border, problems in this neighboring country influence not only United States border towns but also general patterns of immigration into the U.S. and the politics of American entitlement programs. The advent of the North American Free Trade Agreement (NAFTA) agreement in 1994 and severe economic programs in Mexico makes this program particularly appropriate. A training interaction with Mexico is especially timely now since training opportunities for Mexican nationals are severely limited due to the devaluation of the peso against the dollar to nearly the lowest level on record, and on a political situations that closed the doors of the National University, UNAM, for nearly one year. Likewise, due to the fall of a military dictatorship, Chile is enjoying a period of economic and political stability. Chile offers a unique opportunity due to the significant level of training of its scientists and the recent indication of the Chilean federal government's commitment to science and infrastructure development. We have received letters of cooperation from Mexican and Chilean institutions and will continue to have access to trainees selected from over 300 hospitals and 200 research institutions. The proposed program dove-tails with existing expertise, funding, and the presence of proven mentors and a working program in the area of reproductive sciences. Our previous funding period was extremely productive and consistent with the goal of the program. Thesaurus Terms: epidemiology, fertility, health science research, international cooperation, public health, training Mexico, South America Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 30-SEP-1995 Project End: 30-APR-2005 ICD: FOGARTY INTERNATIONAL CENTER IRG: ZHD1
Grant Number: 5P51RR000163-44 PI Name: DORSA, DANIEL M. PI Email: dorsad@ohsu.edu PI Title: Project Title: REGIONAL PRIMATE RESEARCH CENTER Abstract: This 5-year renewal for Grant P51 RR00163 includes requests for support of the Center's AIDS-related research programs and improvement and Modernization projects. The application reflects the significant changes that have occurred at the Center since it was last reviewed in 1994, as well as our plans for expanding the use of nonhuman primates (NHPs) as models for human disease. The aims of this application are to: 1) provide the infrastructure support necessary to conduct advanced research for which NHPs are uniquely suited for solutions of human health problems; 2) enhance the Center's resources (scientific expertise, laboratories, animal services and equipment so as to serve as a regional, national and international resource research through support of our unique programs to develop genetically identical animals, to produce SPF animals free of viruses such as herpes B, and to define animal parentage and genotype; and 4) build new research programs utilizing NHPs in functional genomics, aging, gene therapy and vaccine development. These aims facilitate the research programs housed n the research divisions. Research in the Division of Neuroscience focuses n neuroendocrinology, the effects of aging on brain function, etiology of stress-related pathologies and affective disorders, growth factors and lung development, drug addiction and depression, dietary components is involved in research that includes molecular aspects of CMV latency, blood brain barrier dysfunction in AIDS, studies of SIV neurotropism, effects of altered cell tropism on the pathologic potential of type D retrovirus, the role of T and B cell responses in retrovirus infections, an the role of homocysteine in atherosclerosis. The Division of Reproductive Sciences houses projects including nuclear embryos transfer, causes of premature labor, neuroendocrine regulation of the menstrual cycle and prolactin secretion, the role of sex steroids in ovarian function and vascular occlusion, hormonal control of the reproductive tract, and the cell biology of fertilization. Support services for the intra- and extramural research programs are provided by three services divisions. The Division of Administration oversees all of the operations of the Center. The Division of Animal Resources is responsible for the veterinarian and animal care that supports the health and well being of our animal population. The Division of Information Technology and Engineering oversees all of the communications and bioengineering services of the Center. Thesaurus Terms: Primate, animal colony
Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-MAY-1978 Project End: 30-APR-2004 ICD: NATIONAL CENTER FOR RESEARCH RESOURCES IRG: RIRG
Grant Number: 5R01AG019914-03 PI Name: URBANSKI, HENRYK F. PI Email: urbanski@ohsu.edu PI Title: ASSOCIATE PROFESSOR Project Title: Effect of Aging and Caloric Restriction on Circadian Ph* Abstract: DESCRIPTION (provided by applicant): Human physiology shows strong rhythmic components with peaks and nadirs occurring regularly at specific times of the day. Accumulating evidence suggests that perturbation of the circadian neuroendocrine circuitry may play an important role in several aging-related disorders and may have a major influence on life span. Furthermore, there is growing support for the view that neuroendocrine aging may be caused by oxidative injury, stemming from a progressive overload of a cell's antioxidant capacity. The proposed research will use male rhesus monkeys to examine the mechanism by which caloric restriction can prevent aging-related deterioration of key neuroendocrine circadian rhythms, including DHEAS, cortisol, melatonin and testosterone. We will perform experiments to examine the aging-related progression of circadian dysfunction in ad libitum-fed and age-matched calorie-restricted animals in vivo. This will involve repeated serial blood collections via a catheter-swivel-tether set up, together with concomitant body temperature recordings and actography. In addition, changes in body composition and brain morphology will be assessed non-invasively using DEXA and MRI respectively. Ultimately, the brains of these animals will be examined postmortem to test the hypothesis that caloric restriction protects the aging primate hypothalamus from oxidative injury. Using electron microscopy, we will examine the synaptology of key hypothalamic nuclei involved in mediating circadian neuroendocrine function and use in situ hybridization histochemistry to assess changes in gene expression. We will also use immunohistochemistry to examine loss of specific axonal projections and the associated increase in gliosis. We expect to show that caloric restriction helps to protect hypothalamic nuclei from oxidative injury and, more importantly, that it helps to maintain the integrity of the neural pathway that links the suprachiasmatic nucleus (the central biological clock) to the paraventricular nucleus (the hypothalamic regulator of the adrenal axis). A deeper understanding of aging-related changes in central neuroendocrine circadian circuits of primates and the protective influence of caloric restriction should help to elucidate the mechanism of human aging and help with the development of effective therapies for a wide range of disorders in the elderly. Thesaurus Terms: aging, caloric dietary content, circadian rhythm, dietary restriction, hypothalamus, neuroprotectant, oxidative stress, suprachiasmatic nucleus antioxidant, cortisol, dehydroepiandrosterone, gene expression, melatonin, morphology, testosterone Macaca mulatta, blood chemistry, computer data analysis, confocal scanning microscopy, immunocytochemistry, in situ hybridization, magnetic resonance imaging, male, organ culture, photon absorptiometry, radioimmunoassay Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-SEP-2001 Project End: 31-AUG-2005 ICD: NATIONAL INSTITUTE ON AGING IRG: ZAG1
Grant Number: 5R01AI042490-06 PI Name: GRAVETT, MICHAEL G. PI Email: gravettm@ohsu.edu PI Title: CHIEF Project Title: Experimental Model for Chorioamnionitis and Prematurity Abstract: Prematurity is the leading cause of neonatal morbidity and mortality in the United States. Intrauterine infections are an important, and potentially treatable cause of prematurity, and are associated with increased risk of neonatal white matter lesions of the brain and cerebral palsy. However, the mechanisms by which infection leads to prematurity and/or cerebral palsy remain speculative and treatment strategies untested largely because humans cannot be longitudinally studied following infection. We propose to use chronically instrumented pregnant rhesus monkeys at 120-130 day gestation with experimental intrauterine infection, as previously described (Gravett et al, Am J Obstet and Gynecol; 171:1660-1667,1994) to study the temporal and quantitative relationships among infection, cytokines, prostaglandins, steroid hormones, cytokine antagonists, preterm labor, and neonatal white matter lesions of the brain in order to develop effective interventional strategies. After postoperative stabilization in a tether, we will; (1) inoculate Group B Streptococci (GBS) into the amniotic fluid to establish intrauterine infection and preterm labor. Uterine contractility will be continuously monitored and periodic samples of amniotic fluid and maternal and fetal blood (1-4 cc) will be obtained for assays of eicosanoids, steroid hormones, cytokines, matrix metalloproteinases and for microbial studies; (2) utilize antibiotics with and without potent inhibitors of proinflammatory cytokine production (dexamethasone,IL-10) o prostaglandin production (indomethacin) to ascertain the most effective intervention to down-regulate the cytokine/prostaglandin cascade and associated uterine activity; (3) infuse proinflammatory cytokine IL-1beta into the amniotic cavity through indwelling catheters in the absence of infection. Prior to infusion of IL-1beta in the absence of infection, specific novel proinflammatory cytokine inhibitors (IL-1ra and sTNF-R1 PEG) will be used to identify other potentially useful immunomodulators. Samples of the decidua, fetal membranes, tissues, and brain will be obtained at cesarean section for microbiologic, histopathologic studies, immunohistochemistry for cytokines, localization and quantitation of mRNA for cytokines and PGHS-2. Fetal brain will be examined for increased apoptosis associated with white matter lesions. Leukocytes in amniotic fluid and tracheal aspirates will be assessed by flow cytometry Postpartum, the mother will be treated with appropriate antibiotics to eradicate the GBS from the genital tract and returned to the colony. These studies will clarify the pathophysiology of infection-associated preterm labor and will suggest effective interventional strategies. Thesaurus Terms: antibiotic, antiinflammatory agent, cytokine, microorganism disease chemotherapy, nonhuman therapy evaluation, pregnancy disorder chemotherapy, pregnancy infection, premature labor brain disorder chemotherapy, dexamethasone, disease /disorder model, embryo /fetus chemotherapy, embryo /fetus disorder, immunosuppressive, immunotherapy, indomethacin, interleukin 1, prostaglandin inhibitor Macaca mulatta, Streptococcus agalactiae, embryo /fetus cell /tissue, female, histopathology, immunocytochemistry, neuropathology, newborn animal Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-SEP-1997 Project End: 31-MAY-2006 ICD: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES IRG: HED
Grant Number: 5R01CA075922-07 PI Name: WONG, SCOTT W. PI Email: wongs@ohsu.edu PI Title: ASSOCIATE SCIENTIST Project Title: RHESUS HHV 8 HOMOLOGUE IN AIDS RELATED MALIGNACIES Abstract: Accumulating evidence indicates that human herpesvirus 8 (HHV8), also known as Kaposi's sarcoma-associated herpesvirus (KSHV), is the etiological agent associated with the development of Kaposi's sarcoma (KS). In addition to KS, HHV8 is also associated with the development of primary effusion lymphoma (PEL) and some cases of multicentric Castleman's disease (MCD), both B-cell lymphoproliferative disorders (LPD) observed in humans with the acquired immunodeficiency syndrome (AIDS) also presenting with KS. Understanding how HHV8 is involved in these diverse disease manifestations is complicated with the lack of an accessible animal model that recapitulates these diseases. Recently, the investigators and others reported that rhesus macaques harbor a simian herpesvirus that is closely related to HHV8, referred to as rhesus rhadinovirus (RRV). Moreover, experimental inoculation of rhesus macaques with RRV strain 17577 indicates that in macaques previously inoculated with SIVmac239, RRV strain 17577 induces disease manifestations that possess features that resemble KS and MCD. The long-term objectives of this application are to further elucidate how SIV and RRV induce these disease manifestations and to develop this as an animal model to address HHV8-associated disease. To accomplish this, the following specific aims are proposed. Specific Aim 1: Role of immunosuppression in RRV strain 17677-associated disease. Rhesus macaques will be immunosuppressed by either SIVmac239 infection or by iatrogenic agents and inoculated intravenously with RRV strain 17577. Experimentally inoculated macaques will be monitored for viral load by semi-quantitative PCR, changes in leukocyte subsets in peripheral blood and lymph nodes, serological response to RRV, and for clinical signs of RRV-associated disease. Host response to virus infection will be analyzed for inflammatory cytokine expression and activity by RT-PCR, enzyme-linked immunosorbent assays (ELISA) and immunohistochemical staining on samples collected from peripheral blood, lymph node and retroperitoneal fibromatosis (RF) samples from RRV-, SIVmac239- and RRV/SIVmac239-infected samples. Differential expression of activity of inflammatory responses may indicate how RRV/SIVmac239 infection results in the development of RRV-associated disease. Specific Aim 2: Molecular genetic manipulation of the RRV genome. The investigators propose to initiate molecular genetic manipulation of the RRV genome to identify viral determinants of pathogenesis in SIVmac239-infected rhesus macaques. Viral orfs that they believe are important based upon in vitro analysis of infected tissue samples and in vitro cell culture systems will be targeted and nonsense mutations inserted to abrogate the expression of the viral gene product. Specific Aim 3: In vivo pathogenesis of mutant RRVs with defined nonsense mutations in targeted viral orfs. The results from these proposed studies should provide new insights into how RRV and SIV together induce RRV-associated disease, and serve as a means to elucidate the role of HHV8 in HHV8-associated disease. Thesaurus Terms: Gammaherpesvirinae, Herpesviridae disease, Kaposi's sarcoma, simian AIDS, viral carcinogenesis cytokine, disease /disorder model, gene expression, viremia, virus antigen, virus genetics, virus infection mechanism, virus virus interaction Macaca mulatta, enzyme linked immunosorbent assay, human tissue, immunocytochemistry, polymerase chain reaction Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-JUL-1997 Project End: 30-JUN-2005 ICD: NATIONAL CANCER INSTITUTE IRG: ZRG1
Grant Number: 5R01EY013199-03 PI Name: NEURINGER, MARTHA D. PI Email: neuringe@ohsu.edu PI Title: PROFESSOR Project Title: Dietary Fatty Acids and Photoreceptor Function Abstract: DESCRIPTION (provided by applicant): Docosahexaenoic acid (DHA, 22:6n-3) is a long-chain, highly polyunsaturated n-3 fatty acid. It accounts for about 5O percent of the fatty acids within photoreceptor outer segment disk membranes, the site of phototransduction. Retinal function is altered by changes in retinal membrane DHA content. Rhesus monkeys fed low levels of n-3 fatty acids during development had reduced retinal DHA, impaired visual acuity development and several changes in the electroretinogram (ERG), including prolonged recovery of isolated rod photoreceptor responses to bright, a-wave-saturating flashes. Human infants fed formulas lacking DHA, or containing low levels of its precursors, also showed impairments in the ERG and visual acuity development. In addition, low DHA levels are found in the blood and tissues of many human patients with retinitis pigmentosa and in animal models of retinal degeneration, and dietary DHA is being tested as a possible therapeutic treatment. Despite these indications of DHAs importance in the retina, the nature of the changes in retinal function associated with altered fatty acid composition have not been explored in detail and little is known about their underlying mechanisms. We plan to examine several aspects of retinal function in rhesus monkeys raised on diets with different levels of n-3 fatty acids known to result in widely differing retinal DHA levels. A series of recently developed non-invasive ERG methods will allow us to define effects on phototransduction and photoresponse recovery as well as on rhodopsin regeneration. Specific aims include testing the effects of DHA status on: (1) the activation and deactivation kinetics of isolated rod photoresponses over a wide range of flash intensities; (2) the desensitization of rod responses by background light; and (3) the time-course of dark adaptation over a range of bleaches. In addition, aim 4 will be to test whether any of these alterations in photoreceptor function are reversible after dietary DHA repletion. These studies will provide new information about the modification of retinal function, particularly adaptive processes, by membrane DHA content. Together with in vitro studies of model membranes, these data will help to provide the basis for hypotheses about the underlying processes that are altered by differing retinal fatty acid compositions. They also will contribute to understanding the changes in retinal function found in human infants fed diets lacking DHA or its precursors. Thesaurus Terms: dark adaptation, dietary lipid, light adaptation, omega 3 fatty acid, visual photoreceptor, visual phototransductiondietary supplement, fluidity, malnutrition, membrane activity, rod cell, visual perception Macaca mulatta, electroretinography, nutrition related tag Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-AUG-2001 Project End: 31-JUL-2004 ICD: NATIONAL EYE INSTITUTE IRG: VISC
Grant Number: 5R01HD014643-21 PI Name: SMITH, M SUSAN. PI Email: smithsu@ohsu.edu PI Title: ASSOCIATE PROFESSOR OF PHYSIOLOGY Project Title: CONTROL OF GONADOTROPIN SECRETION DURING LACTATION Abstract: The central hypothesis of this proposal is that the suppression of GnRH neuronal activity during lactation is due to neural impulses, derived from the suckling stimulus, that alter hypothalamic function and to the change in energy balance associated with milk production. We have identified specific areas in the brainstem that are activated by the suckling stimulus; neurons from each of these areas send projections to the arcuate nucleus of the hypothalamus. We also reported that several neuronal systems in the arcuate nucleus are altered during lactation (increased NPY and AGRP, decreased POMC). These changes are consistent with the chronic hyperphagia of lactation. Our studies showed that NPY projections from the arcuate nucleus makes direct contact with GnRH neurons in the preoptic area and with CRH neurons in the periventricular nucleus (a key site for regulation of food intake). Also, NPY receptors (Y5 subtype) are expressed on GnRH and CRH neurons. Thus, we have established the neuroanatomic framework by which increased NPY activity in the arcuate nucleus could serve as a key element in linking changes in energy balance to the suppression of GnRH neuronal activity during lactation. Another indicator of the change in energy balance is the suppression of leptin during lactation in association with milk production. The proposed experiments expand on these findings and will use three approaches: 1) Neuroanatomical studies will determine the phenotypes of the suckling-activated brainstem neurons that make contact with NPY or POMC neurons in the arcuate nucleus and with GnRH neurons in the preoptic area. 2) Physiological studies will determine if the increase in NPY and the decrease in leptin play functional roles in the suppression of GnRH neuronal activity. 3) Functional genomics will be used to identify additional relevant genes that play key roles in the regulation of NPY and GnRH neurons and in conveying information about the state of energy balance during lactation. The interaction between reproductive function and energy balance during lactation provides a physiological model for studying a number of conditions in women (under-nutrition, anorexia nervosa, bulimia and exercise-induced amenorrhea) that involve a suppression of reproductive function associated with changes in energy balance. All of these conditions have common mechanisms underlying the decrease in GnRH activity. Thesaurus Terms: gonadotropin releasing factor, hormone regulation /control mechanism, lactation, neuroanatomy, neuron, neuropeptide Y bioenergetics, brain stem, gene expression, hypothalamus, leptin, luteinizing hormone, phenotype, protein structure function, secretion, stimulus /response confocal scanning microscopy, immunocytochemistry, immunofluorescence technique, in situ hybridization, injection /infusion, laboratory rat, microarray technology, radiotracer, tissue /cell culture Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-DEC-1979 Project End: 28-FEB-2005 ICD: NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT IRG: REN
Grant Number: 5R01HD019182-18 PI Name: BRENNER, ROBERT M. PI Email: brennerr@ohsu.edu PI Title: SENIOR SCIENTIST Project Title: HORMONAL CONTROL OF THE FEMALE REPRODUCTIVE TRACT Abstract: DESCRIPTION: (Adapted from the applicant's abstract) Much remains to be learned about the periodic menstrual sloughing, bleeding and regenerative repair process so characteristic of the primate endometrium. During the previous research period, the investigator discovered an unexpected involvement of vascular endothelial growth factor (VEGF) and its receptors (R) in the menstrual inductive process. Very early in the premenstrual phase, one of the VEGF receptors, VEGFR-2, also known as KDR, was strongly expressed in the stromal cells (not the vascular endothelium) of the upper endometrial zones, the same cell population that expresses most of the matrix metalloproteinases (MMPs). MMPs are enzymes responsible for the tissue destruction and sloughing associated with menstruation. FLT-1, another VEGF receptor, was also transiently upregulated in the glandular cells which also produce an MMP, called matrilysin. Therefore the investigators hypothesize that VEGF acts through KDR and FLT-1 to facilitate MMP expression in the upper endometrial zones during menstrual induction. Hepatocyte growth factor (HGF) was also expressed transiently in the same endometrial stromal cells at the same time. The investigators hypothesize that HGF and VEGF may interact during the LFT to facilitate MMP upregulation. Keratinocyte growth factor (KGF), a growth factor strongly upregulated by P, had strong trophic effects on the spiral arteries, but the KGF receptor was not detectable in the arteries. The arteries do express KDR and FLT-1, so they hypothesize that KGF acts indirectly through the VEGF system to mediate spiral artery growth under P influence. In the current application they propose to explore the interactions between VEGF, HGF and KGF in the primate endometrium within the following specific aims: 1. The VEGF family and MMPs during menstrual induction. 2. VEFG-HGF interactions in MMP regulation. 3. KGF-VEGF interactions in spiral artery growth. 4. HGF and VEGF antagonists during menstruation. Thesaurus Terms: fibroblast growth factor, growth factor receptor, hepatocyte growth factor, hormone regulation /control mechanism, menstrual cycle, metalloendopeptidase, progesterone, receptor expression, vascular endothelial growth factor cell cycle, cervix, endometrium, fallopian tube, inhibitor /antagonist, protein protein interaction, receptor binding DNA footprinting, Macaca mulatta, RNase protection assay, female, hormone therapy, immunocytochemistry, morphometry, northern blotting, tissue /cell culture Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-SEP-1984 Project End: 30-APR-2005 ICD: NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT IRG: REB
Grant Number: 5R01HD020869-17 PI Name: STOUFFER, RICHARD L. PI Email: stouffri@ohsu.edu PI Title: SENIOR SCIENTIST AND HEAD Project Title: PROGESTERONE RECEPTOR AND ACTION IN THE PRIMATE OVARY Abstract: DESCRIPTION: (Adapted from the applicant's abstract) The demonstrated activity of progestin to promote ovulation, coupled with the demonstration of progesterone receptor in the luteinizing, ovulating follicle and in the differentiated corpus luteum is compelling evidence for further investigation of the potential actions of progesterone in the ovulating follicle and corpus luteum. This proposal is directed to an investigation of the molecular and cellular events that are steroid/progesterone regulated during follicle rupture and luteinization of the periovulatory follicle, in the maintenance of luteal structure-function in the developed corpus luteum of the nonfertile cycle, and in the rescue of the corpus luteum during early pregnancy. Three treatment protocols will be used in which luteotropic support is sustained via exogenous luteinizing hormone/chorionic gonadotropin during the periovulatory interval, at midluteal phase of the cycle, and in simulated early pregnancy. These protocols are combined with progesterone ablation (using the 3B-hydroxysteroid dehydrogenase inhibitor, Trilostane) and progestin replacement (using the analog, R5020), followed by removal of periovulatory follicle or corpus luteum for analyses. These analyses include indices of tissue remodeling (protease expression, vascularization, cell proliferation), tissue differentiation (cholesterol utilization, steroid genesis, steroid receptor expression), and tissue regression (apoptosis). From these approaches, novel autocrine actions of progesterone in the luteinizing follicle and corpus luteum may be revealed. Thesaurus Terms: corpus luteum, graafian follicle, hormone regulation /control mechanism, ovary, ovulation, progesterone, progesterone receptor, receptor expression apoptosis, cell proliferation, chorionic gonadotropin, estradiol, luteinizing hormone, menstrual cycle, messenger RNA, pregnancy, prostaglandin, prostaglandin E, prostaglandin F Macaca mulatta, immunocytochemistry, in situ hybridization, northern blotting, polymerase chain reaction, western blotting Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-JUL-1985 Project End: 28-FEB-2006 ICD: NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT IRG: REB
Grant Number: 5R01HD024870-16 PI Name: OJEDA, SERGIO R. PI Email: ojedas@ohsu.edu PI Title: SCIENTIST/DIVISION HEAD Project Title: NEURAL CONTROL OF THE PREPUBERTAL OVARY Abstract: DESCRIPTION: (Adapted from the applicant's abstract) This is a renewal application aimed at elucidating some of the basic neuroendocrine and cellular mechanisms overning mammalian ovarian development. During the last period of support we employed a combination of cellular, molecular and genetic approaches to demonstrate the existence of a neurotrophin-mediated regulatory system that is acting at the interface between the endocrine and nervous systems -- contributes to the developmental control of ovarian function. In related studies, we identified a series of additional genes that may belong to the hierarchy of regulatory molecules controlling ovarian folliculogenesis, follicular growth and ovulation. The recognition of these new regulatory components, and the use of genetic approaches to modify the expression of genes in a cell-specific and temporally restricted manner, provides us with a new opportunity to unravel some of the key cell-cell regulatory mechanisms underlying mammalian ovarian development. To initiate this undertaking we propose two sets of studies: The first, to define the physiological importance of NGF and its receptors I follicular growth and ovulation, and their contribution to the etiology of ovarian cystic disease; the other, to elucidate the role that a set of functionally diverse genes found to be differentially expressed in the ovary at two key developmental phases (folliculogenesis and first ovulation) may play in the regulation of these processes. To this end, the following specific aims are proposed: 1. To test the hypothesis that, while required for normal follicular growth, NGF overproduction in endocrine cells of the follicular wall leads to the development of cystic ovarian disease via activation of the low-affinity neurotrophin receptor, p75NGFR. 2. To examine the hypothesis that activation of trkA, the high ffinity tyrosine kinase NGF receptor, in ovarian endocrine cells of mesenchymal origin contributes to the completion of two critical phases in the natural history of the developing ovary, the initiation of preantral follicular development and rupture of the follicular wall at ovulation. 3. To define the role that insulin receptor-related receptor (IRR), an orphan receptor of the insulin receptor family recently found to be co-expressed with trkA in thecal cells of periovulatory follicles, plays in ovulation, and to molecularly identify the gene encoding the IRR ligand. 4. To test the hypothesis that follicular formation requires the temporal and cell-specific coordinated expression of three functionally diverse genes found to be differentially displayed at the time of folliculogenesis; the adhesion molecule Cadherin-11, the protooncogene PTTG, and the transcriptional regulator ENX-1. Thesaurus Terms: animal puberty, cell differentiation, growth factor receptor, histogenesis, nerve growth factor, neuroendocrine system, neuroregulation, ovary, protein structure function, receptor expression cadherin, developmental genetics, gene expression, graafian follicle, insulin receptor, ovulation, pathologic process, polycystic ovary syndrome, protein tyrosine kinase, protooncogene, transcription factor gene targeting, laboratory rat, transgenic animal Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-FEB-1988 Project End: 31-JAN-2006 ICD: NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT IRG: REB
Grant Number: 5R01HD025123-14 PI Name: OJEDA, SERGIO R. PI Email: ojedas@ohsu.edu PI Title: SCIENTIST/DIVISION HEAD Project Title: NEUROENDOCRINOLOGY OF PUBERTY AND SEXUAL DEVELOPMENT Abstract: This is a renewal application aimed at elucidating the neuroendocrine mechanisms involved in controlling the initiation of mammalian puberty. During the current period of support, our studies have: a) provided evidence for the existence of key signaling molecules utilized by glial cells to regulate the secretory activity of luteinizing hormone releasing hormone (LHRH) neurons, b) unveiled the existence of a higher level of hierarchy in the neuroendocrine cascade that controls the onset of puberty and c) identified a potential new component of the hypothalamic regulatory complex controlling females sexual development. We now propose the use of two different conditional gene targeting approaches to disrupt the function of these newly recognized regulatory molecules in a cell-specific and temporally-restricted manner, and thus, test the hypothesis that they are essential components of the central mechanism controlling the acquisition of female reproductive capacity. To this end, the following specific aims are proposed: 1. To test the hypothesis that an astrocyte-specific, temporally-controlled disruption of erbB-1 receptors, which mediate the actions of transforming growth factor alpha (TGFalpha), delays female sexual maturation by affecting both the gonadal-independent and steroid- dependent activation of LHRH release. 2. To examine the hypothesis that selective disruption of astroglial erbB-2 coreceptors, which are required for amplification of hypothalamic erbB-1-and erbB-4-mediated actions, delays sexual maturation when effected at key phases of LHRH neurosecretory activity. 3. To investigate the hypothesis that selective disruption of astroglial erbB-4 receptors, which mediate the actions of NRGs in hypothalamic astrocytes, results in maturational deficits similar to (or more pronounced than) those caused by the loss of erbB- 1/erbB-2-mediated signaling. 4. To define the role that Nel, a recently identified neuronal protein with EGF-like repeats, may play in the cell-cell communication process underlying the hypothalamic control of sexual development. 5. To test the hypothesis that TTF-1, a member of the Nkx family of homeodomain genes that remains postnatally expressed in discrete hypothalamic regions, is an intrinsic component of both the neuron-to-neuron and glia-to-neuron signaling process controlling the onset of female puberty. Thesaurus Terms: animal puberty, gonadotropin releasing factor, luteinizing hormone, neuroendocrine system, transforming growth factor astrocyte, developmental genetics, epidermal growth factor, estradiol, gene expression, growth factor receptor, homeobox gene, hormone regulation /control mechanism, hypothalamus, protein tyrosine kinase, receptor expression female, gene targeting, laboratory mouse, laboratory rat, transgenic animal Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-SEP-1990 Project End: 31-MAY-2005 ICD: NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT IRG: BCE
Grant Number: 5R01HD029186-06 PI Name: URBANSKI, HENRYK F. PI Email: urbanski@ohsu.edu PI Title: ASSOCIATE PROFESSOR Project Title: Postnatal Development of Ionotropic Glutamate Receptors Abstract: L-glutamate is the most abundant excitatory neurotransmitter in the mammalian central nervous system. It plays a major role in behavioral processes such as learning and memory, and is a key component of the neuroendocrine mechanism that controls sexual maturation. Although ionotropic glutamate receptors have been studied extensively in the rodent brain, both at the molecular and pharmacological levels, the postnatal ontogeny of these receptors in humans is poorly understood. To help resolve this issue, the proposed studies will use sexually immature male and female rhesus macaques (Macaca mulatta) to test various hypotheses regarding: 1) the temporal expression of genes encoding different glutamate receptor subunits, 2) the neurotransmitter identity of neurons that show developmental plasticity in glutamate receptor gene expression, and 3) the influence of the changing sex steroid environment on the induction of these developmental receptor changes. The studies will involve a series of molecular and immunohistochemical approaches to characterize and quantify glutamate receptor gene expression at three key stages of postnatal development: 1) infantile, 2) juvenile, and 3) peripubertal, both with and without experimental manipulation of circulating estradiol and testosterone concentrations. Because macaques and humans show similar postnatal cognitive developments and similar developmental changes in their sex-steroid environment, the proposed studies are expected to yield new information about the ontogeny of ionotropic glutamate receptors in humans. Moreover, because the subunit composition of different glutamate receptors determines their affinity for different ligands (e.g., NMDA, AMPA and kainate) and influences their functional properties (e.g., permeability to Ca2+), elucidation of the mechanisms that regulate their developmental expression should help to lay a foundation for the development of pharmacological treatments for pediatric neurological disorders. Thesaurus Terms: brain mapping, developmental genetics, developmental neurobiology, gene environment interaction, glutamate receptor, hormone regulation /control mechanism, neuron, neurotransmitter, protein structure function, sex hormone age difference, animal puberty, estradiol, gender difference, infant animal, juvenile animal, luteinizing hormone, neural plasticity, neuroendocrine system, neurogenetics, testosterone Macaca mulatta, RNase protection assay, immunocytochemistry, in situ hybridization, ovariectomy Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-SEP-1993 Project End: 31-MAR-2007 ICD: NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT IRG: BCE
Grant Number: 5R01HD037131-05 PI Name: SPINDEL, ELIOT R. PI Email: spindele@ohsu.edu PI Title: SENIOR SCIENTIST Project Title: FETAL NICOTINE EXPOSURE EFFECT ON PRIMATE LUNG Abstract: The deleterious effects of maternal smoking during pregnancy are all too well established. Maternal smoking is the major preventable cause of intrauterine growth retardation and prematurity. Perhaps less well appreciated, is the recent, overwhelming evidence, that smoking during pregnancy directly and adversely effects lung development. Respiratory problems associated with in utero tobacco exposure include decreased lung function, increased respiratory diseases and increased incidence of sudden infant death syndrome (SIDS). Given the unfortunate prevalence of smoking during pregnancy and the resulting serious consequences, it is of major importance to understand the mechanisms underlying smoking-induced changes in the newborn. Our preliminary data suggests that nicotine itself is one of the factors responsible for the changes in pulmonary function observed in neonates born to smoking mothers. In this application we propose to use the rhesus monkey to characterize the effects of chronic exposure to low levels of nicotine throughout pregnancy on lung development and subsequent pulmonary function. Whole animal studies will be complemented with in vitro studies to begin to determine the molecular mechanisms underlying nicotine's effect on lung. In preliminary studies we have demonstrated that exposure of pregnant rhesus monkeys to a nicotine dose consistent with that of smokers alters fetal airway development and that related effects can be produced in fetal monkey lung organ cultured. Immunohistochemistry shows wide expression of nicotinic receptors in developing lung and nicotine appears to alter the pattern of receptor expression. Preliminary data further suggests that some of the effects of nicotine, acting through nicotinic receptors, may be mediated by antagonism of the mitogenic effects of peptide growth factors. Thus we specifically propose to 1, Determine the basis for nicotine's actions by determining the time course and cell specific expression of nicotinic receptor subtype expression in fetal monkey lung; 2, Characterize the effect of fetal exposure to nicotine on lung development and function by functional, morphometric, immunohistochemical and molecular analysis; and 3, begin to determine the mechanism underlying nicotine's actions by use of fetal monkey lung organ culture. From these studies will come the first description of the effects of chronic nicotine exposure on lung function; a determination of the extent to which these effects are reversible; and a beginning understanding of the mechanisms underlying these effects. Definitive knowledge of the effects of nicotine on lung development would provide an important additional tool in smoking control and will begin to better explain the link between maternal smoking and altered neonatal respiratory function. Thesaurus Terms: embryo /fetus toxicology, histogenesis, lung, lung development, nicotine, respiratory function biological model, gastrin releasing peptide, nicotinic receptor, receptor expression Macaca mulatta, immunocytochemistry, in situ hybridization, organ culture, polymerase chain reaction Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-FEB-1999 Project End: 31-JAN-2004 ICD: NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT IRG: LBPA
Grant Number: 5R01HD042000-02 PI Name: HENNEBOLD, JON D. PI Email: henneboj@ohsu.edu PI Title: Project Title: Epoxyeicosatrienoic Acids in the Ovulatory Follicle Abstract: Limited published reports and preliminary experiments conducted by the PI suggest that the rodent and primate ovary possess the capacity to produce and metabolize epoxyeicosatrienoic acids (EETs), metabolites of arachidonic acid (AA). Preliminary experiments also suggest that EETs play a role in ovulation as an inhibitor of EET metabolism significantly increased the number of oocytes released in mice following injection of an ovulatory dose of gonadotropin. Therefore, studies are designed using rodents and primates to test the hypothesis that EETs are synthesized (Aim number 1), inactivated (Aim number 2), and directly affect molecular processes (Aim number 3) and oocyte release (Aim number 4) in the ovulatory follicle. Expression of EET generating cytochrome P450 (CYP) epoxygenases during the periovulatory interval and their cellular localization will be determined by real-time PCR and in situ hybridization, respectively. The specific EET isomer(s) produced during the periovulatory interval will be determined by gas chromatography/mass spectrometry. Recently, the PI has cloned a novel ovary-selective isoform of soluble (cytoplasmic) epoxide hydrolase (sEH), an enzyme that converts EETs to their inactive diols (dihydroxyeicosatrienoic acids; DiHETEs). The expression of the ovary-selective sEH isoform was restricted to the periovulatory period. Biochemical properties of the novel ovary- selective sEH will be determined, including substrate specificity, KM/Vmax, capacity to metabolize EETs and subcellular localization. The presence of a homologous primate isoform(s) will be analyzed by rapid amplification of cDNA ends (RACE). Recent studies have demonstrated that in non-ovarian cells EETs increase the expression of prostaglandin H synthase-2 (PGHS-2), a prostaglandin (PG) synthetic enzyme critical for optimal ovulatory efficiency. The major EET species produced in the ovary during the periovulatory interval will be analyzed in vitro with respect to their ability to directly induce PGHS-2 expression. In vivo effects on PGHS-2 expression, PG production, and ovulation will be determined by using inhibitors of EET generation and metabolism. These studies will provide insight into the action and regulation of EETs in the ovulatory follicle and may yield novel approaches for the regulation of fertility. Thesaurus Terms: cytochrome P450, eicosa 5,8,11 trienoate, eicosanoid metabolism, enzyme activity, epoxide hydrolase, graafian follicle, ovulation, prostaglandin endoperoxide synthase enzyme substrate, estrus, gene expression, granulosa cell, isomer, isozyme, luteinizing hormone, ovulation time, prostaglandin E, prostaglandin receptor Macaca mulatta, egg /ovum, gas chromatography mass spectrometry, immunocytochemistry, in situ hybridization, laboratory mouse, polymerase chain reaction, tissue /cell culture, western blotting Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-JUN-2002 Project End: 31-MAY-2007 ICD: NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT IRG: REN
Grant Number: 5R01HD043209-02 PI Name: BRENNER, ROBERT M. PI Email: brennerr@ohsu.edu PI Title: SENIOR SCIENTIST Project Title: Suppression of Breakthrough Bleeding in LNG-IUS Users Abstract: DESCRIPTION (provided by applicant): The Mirena(c) (Leiras, OY, Finland) is an intrauterine system (IUS) that releases levonorgestrel (LNG) and provides women with highly effective contraception for 5 years. However, serious breakthrough bleeding (BTB) can occur that leads many patients to discontinue treatment. In women using subcutaneous Norplant contraception, BTB can be suppressed by intermittent antiprogestin therapy with mifepristone. NICHD has a new antiprogestin, CDB-2914, that is more potent than mifepristone. The goal of this proposal is to determine whether CDB-2914 can suppress the BTB associated with the LNG-IUS. Preclinical studies will be done first in rhesus macaques fitted with an LNG-IUS and the information gained will be transmitted to Professor Hilary Critchley, University of Edinburgh, who will select a test population of 150 women from a pool of over 400 who are being fitted annually with an LNG-IUS for contraception. The bleeding-suppressive dose and schedule of CDB-2914 that works well in macaques will be adapted for use in women. The work consists of two major aims: Aim 1: to establish an effective, bleeding-suppressive dose schedule of CDB 2914 in macaques, and to assess spatio-temporal expression of bleeding-associated factors in the endometrium. Aim 2: to conduct a randomized, placebo controlled trial to evaluate CDB-2914 suppression of BTB in women being fitted with the LNG-IUS; and for added value, to develop a questionnaire to assess acceptability of the proposed treatment to women. This proposal involves translation of the information gained from basic research with macaques into clinical practice in women within the time frame of the grant. It is essential to conduct the clinical work at the University of Edinburgh, Scotland, UK, because no clinic in the USA has such a large patient population of women being fitted with the LNG-IUS for contraception. The macaque model provides excellent predictability for human studies, and Drs. Brenner and Critchley have published collaboratively on data from macaques and women in several recent papers. The proposal brings basic scientists and clinicians together for a "bench to bedside" approach that will advance women's health and improve contraceptive technology. Thesaurus Terms: drug adverse effect, endometrium, hemorrhage, hormone inhibitor, human therapy evaluation, levonorgestrel, nondrug contraceptive, progestin, reproductive system disorder chemotherapy clinical trial, dosage, gene expression, nonhuman therapy evaluation, pharmacokinetics, reproductive system pharmacology, women's health British Isles, Macaca mulatta, biopsy, clinical research, female, human subject, microarray technology, morphometry, patient oriented research Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 27-SEP-2002 Project End: 30-JUN-2007 ICD: NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT IRG: ZHD1
Grant Number: 5R01HL066118-02 PI Name: SPINDEL, ELIOT R. PI Email: spindele@ohsu.edu PI Title: SENIOR SCIENTIST Project Title: Nicotine & Alpha7 nicotinic receptor in lung development Abstract: DESCRIPTION (provided by applicant): According to the latest statistics from the CDC in 1999, 12.3% of American women smoked during pregnancy, translating to over 400,000 smoke-exposed infants. Smoking during pregnancy is the largest preventable cause of low birth weight, premature delivery, neonatal morbidity, and mortality. Indeed it has been estimated that 10% of all fetal and neonatal deaths are due to smoking during pregnancy. Perhaps less well appreciated is the recent, evidence that smoking during pregnancy directly and adversely affects lung development as manifested by altered pulmonary function and increased respiratory illness in children born of smoking mothers. Remarkably, how smoking produces these effects is unknown. While the cause of pulmonary damage caused by maternal smoking is likely to be multifactorial, it is the basic hypothesis of this application that part of the effect of maternal smoking on lung is mediated by nicotine transported across the placenta to interact with alpha7 nicotinic receptors in developing lung. Our preliminary evidence indicates 1) that alpha7 nicotinic receptors are highly expressed in developing lung; 2) that prenatal nicotine exposure alters alpha7 nicotinic receptor expression in lung; and 3) that collagen gene expression is markedly up-regulated in areas of altered alpha7. Suggesting that nicotine's effect on collagen is mediated by alpha7 receptors, prenatal nicotine exposure has no effect on collagen gene expression in the lungs of cx7 knockout mice. In exciting preliminary data, nicotine inhibits fibroblast proliferation from cells isolated from wildtype neonatal mouse lung, but has no effect on proliferation of fibroblasts from alpha7 knockout mice. This suggests that some of the growth retardation caused by smoking during pregnancy may be mediated by the interaction of nicotine with alpha7 receptors. In this application, using alpha7 knockout and alpha7 gain of function mice, we propose to first demonstrate a link between the effects of prenatal nicotine exposure and alpha7 nAChR, then using cultured pulmonary fibroblasts and epithelial cells begin to determine the mechanism by which nicotine produces these effects. Based on our preliminary data and epidemiologic data on human infants, we will focus on 3 aspects of smoking's effects on lung development: pulmonary function as measured by active and passive tests, cell growth, and collagen expression. From these studies will come some of the first explanations of the molecular mechanisms that underlie the effects of smoking during pregnancy on lung development. These findings will also potentially point to ways to block some of those effects of smoking during pregnancy as well as assist in fighting smoking during pregnancy. Thesaurus Terms: drug interaction, embryo /fetus toxicology, embryology, lung development, lung injury, nicotine, nicotinic receptor cell proliferation, collagen, drug administration rate /duration, drug administration route, gene expression, placental transfer, receptor expression, respiratory function, tobacco abuse female, gene targeting, laboratory mouse, morphometry, plethysmography, tissue /cell culture, transgenic animal Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-JUL-2002 Project End: 30-JUN-2007 ICD: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE IRG: HED
Grant Number: 5R01MH062677-03 PI Name: BETHEA, CYNTHIA L. PI Email: betheac@ohsu.edu PI Title: SENIOR SCIENTIST Project Title: OVARIAN STEROID REGULATION OF SEROTONIN IN PRIMATES Abstract: DESCRIPTION (applicant's abstract): The previous interest of this laboratory in the neural regulation of progestin-induced prolactin secretion in primates has evolved into a broader interest in the regulation of serotonin neural function by ovarian steroids. The serotonin neural system projects to nearly every area of the forebrain and serotonin plays a major role in the regulation of numerous autonomic and cognitive neural processes. Thus, understanding the action of ovarian steroids in the serotonin neural system has relevance to many aspects of mental health and function in women. This proposal continues our search, at a cellular and molecular level, for neural targets of progesterone (P) which are unique from the action of estrogen (E) and it initiates studies to determine the mechanism by which E exerts differential effects on the expression of 3 pivotal genes: tryptophan hydroxylase (TPH), the serotonin reuptake transporter (SERT), and the 5-HT1A autoreceptor, in serotonin neurons of macaques. The overall hypothesis is that progesterone (P) has unique, undiscovered, genomic actions in the serotonin neural system which elevate 5-HT neurotransmission. Estrogen (E) is required for the induction and maintenance of nuclear P receptors and E alone changes the expression of pivotal genes related to serotonin synthesis, uptake, and neuronal firing. The molecular actions of E may be mediated by ER-beta and could involve a protein-protein interaction with nuclear factor kappa B (NF-kB). Aim 1 will obtain definitive evidence that addition of P to an E regimen increases serotonin neurotransmission by application of microdialysis and measurement of serotonin in the extracellular compartment of terminal fields in steroid-treated macaques. Aim 2 will determine the functional consequences of previously reported changes in TPH, SERT, and 5-HT1A autoreceptor gene expression in serotonin neurons of macaques treated with E and P. Aim 3 will determine the effect of E and P on degradative mechanisms of serotonin. Gene expression and function of monoamine oxidase (MAO-A) will be determined in the dorsal raphe and hypothalamic termina1 field. Aim 4 will seek the expression of ER-beta and determine if nuclear factor kappa B (NF-kB) is co-expressed and regulated by E or P in serotonin neurons of macaques. Aim 5 will use laser capture to obtain pure populations of serotonin neurons from steroid-treated macaques and amplify their RNA for examination of genes related to phosphorylation events. These experiments will (1) further the hypothesis that E and P increase serotonin neural function and (2) initiate investigations of the mechanism of action of E and P in serotonin neurons. Thesaurus Terms: estrogen, hormone regulation /control mechanism, neural transmission, ovary, progesterone, serotonin amine oxidase (flavin), gene expression, limbic system, neuron, nuclear factor kappa beta, nuclear receptor, phosphorylation, serotonin receptor, serotonin transporter, tryptophan 2,3 dioxygenase Macaca, animal genetic material tag, immunocytochemistry, microarray technology, microdialysis, molecular cloning, western blotting Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-FEB-2001 Project End: 30-NOV-2005 ICD: NATIONAL INSTITUTE OF MENTAL HEALTH IRG: ZRG1
Grant Number: 5R01NS044330-02 PI Name: WOLF, DON P. PI Email: wolfd@ohsu.edu PI Title: SENIOR SCIENTIST Project Title: Genetically Modified Rhesus Monkeys Abstract: DESCRIPTION (provided by applicant): Neurogenetic diseases cause tens of thousands of deaths in the United States each year, inflict immeasurable pain and suffering, and consume a substantial portion of scarce healthcare resources. A mouse counterpart for many of these diseases does not exist necessitating the creation and use of new mammalian models. Despite the significant challenges associated with the development of monkey models of neurogenetic diseases, the time is appropriate and the need is compelling. Accordingly, our long-term goal is to produce genetically modified Rhesus monkeys that will serve as models for human neurogenetic diseases. We will focus on three, early-onset, loss of function conditions: Kallmann's syndrome, Lesch-Nyhan's disease and Ataxia-Telangiectasia. We will attempt to establish the paradigm relatively quickly, an objective that can not be met with diseases that require decades to reveal themselves. Because Kallmann's syndrome and Lesch-Nyhan's disease are due to mutations in genes located on the X chromosome (KAL1 and HPRT, respectively), loss of function XY cell mutations require disruption of only one allele. Disruption of the two copies of the autosomal Ataxia Telangiectasia Mutated (ATM) gene, while more difficult, will establish the methods necessary for disrupting autosomal genes in vitro. Our working hypothesis is that gene targeting and somatic cell cloning technology can, in combination, provide the basis for generating a reliable supply of animals that accurately represent human disease. The objective of this application is to create the infrastructure necessary to genetically modify Rhesus monkey cells in culture and to use those cells as donors for nuclear transfer. The resultant viable embryos of the desired genotype can then be transferred into surrogate mothers. Genetically modified Rhesus macaques will result. Such animals should provide a resource for the study of human neurogenetic diseases and serve as pre-clinical models for new experimental treatments including gene and stem cell based therapies. Thesaurus Terms: Macaca mulatta, biological model, model design /development, neurogenetics, transgenic animal Kallmann's syndrome, Lesch Nyhan syndrome, ataxia telangiectasia, gene expression, genotype, hypoxanthine phosphoribosyltransferase, telomerase gene targeting, nuclear transfer, polymerase chain reaction, site directed mutagenesis, tissue /cell culture Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 15-AUG-2002 Project End: 30-JUN-2007 ICD: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE IRG: ZRG1
Grant Number: 5R01RR016030-03 PI Name: WOLF, DON P. PI Email: wolfd@ohsu.edu PI Title: SENIOR SCIENTIST Project Title: PROPAGATION OF MONKEY MODELS OF HUMAN DISEASE Abstract: DESCRIPTION (Adapted from the applicant's abstract): There is currently a significant need for populations of animals with specified genotypes which cannot be satisfied by the importation of animals from the wild or by the identification and propagation of valuable founder animals by selective breeding. Indian-origin, rhesus macaques carrying the MHC class 1 allele, A*01, are particularly needed for vaccine development research. The objective of this application is to demonstrate that ARTs can be applied to the rapid, efficient propagation of a valuable founder animal and to the production of identical twins using existing technology, thereby establishing a new approach to satisfying animal requirements of the biomedical research community. The rationale for focusing on a homozygous, founder male is simple. All offspring produced by this animal will be Mamu-A*01 positive. Moreover, if heterozygous carriers of the A*01 allele are used as oocyte donors, 50% of the offspring will be homozygous for the allele, creating additional founder animals. The applicants have access to a male at the NERPRC that is presumed homozygous for the Mamu-A*01 allele based on the fact that 100% (15 of 15; K. Mansfield, personal communication) of his offspring are Mamu-A*01 positive. The number of sperm in an average ejaculate, when used in conjunction with ICSI, can result in the production of hundreds, if not thousands, of embryos. Sperm collection, preservation during transportation or storage and use of ICSI is established technology in the applicants' laboratory, allowing a high probability of success. The investigators propose to establish a new paradigm for the efficient, cost effective propagation of founder monkeys and populations of valuable, genetically defined macaques, including identical twins, through conduction of the following specific aims: 1) Produce 25 Mamu-A*01 positive animals in the first year and 50 animals per year for the duration of this study using sperm from a homozygous Mamu-A*01 positive donor. This aim will establish the paradigm and create new populations of heterozygous and homozygous animals without impacting the natural reproduction of either the sperm or the oocyte donors. The applicants will confirm homozygosity in candidate animals by creating embryos with their gametes and monitoring the presence or absence of the Mamu-A*01 allele in these embryos; and 2) Employ blastomere separation and culture or embryo splitting technology to increase embryo numbers and to produce genetically identical animals. The applicants will select the optimal approach to twinning in year 1 and produce 5 sets of identical twins per year during years 2-5 of the study. Thesaurus Terms: animal breeding, disease /disorder model, model design /development, monozygotic twin AIDS, AIDS vaccine, HIV infection, MHC class I antigen, allele, egg /ovum, fertility, genotype, heterozygote, homozygote, human immunodeficiency virus, simian immunodeficiency virus, sperm, vaccine development Macaca mulatta, cryopreservation, embryo /fetus culture Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-JUL-2001 Project End: 30-JUN-2006 ICD: NATIONAL CENTER FOR RESEARCH RESOURCES IRG: RIRG
Grant Number: 5R21NS041601-02 PI Name: PAU, KWOK-YUEN F. PI Email: pauf@ohsu.edu PI Title: STAFF SCIENTIST Project Title: Characterization and Differentiation of Monkey ES Cells Abstract: DESCRIPTION (provided by applicant): The application of embryonic stem (ES) cell therapy for the treatment of human degenerative diseases should be performed in nonhuman primates prior to human trials. The macaque is an ideal animal model for neural transplantation studies because the developmental potential of the grafted cells can be timely and precisely examined postmortem. As a prelude to in vivo studies, the developmental potential and cell lineage differentiation of monkey ES cells must be established in vitro. To this end, we propose to culture and characterize monkey ES cells free of feeder cells before differentiating them into high-percentage cholinergic and dopaminergic neurons, and to determine the pluripotency of monkey ES cells by single cell culture and spontaneously differentiation into cells of all three embryonic germ layers in vitro, thereby addressing the objectives of this PA (PA-99-086/-01-076, NOVEL APPROACHES TO ENHANCE STEM CELL RESEARCH). Aim 1 will characterize two monkey ES cell lines with an XX and XY karyotype that are cultured under feeder cell-free conditions. Rhesus monkey ES cells will be plated on Matrigel- or laminin-coated dishes and cultured in a serum-free medium supplemented with fibroblast growth factors (FGFs), laminin, serum replacement, or conditioned medium from cell cultures to sustain the pluripotency and undifferentiated states. The growth rate, karyotype normalcy, and undifferentiated state will be determined. Aim 2 will directionally differentiate monkey ES cells into dopaminergic and cholinergic neurons, and to differentiate single monkey ES cell into all three embryonic germ layers in vitro. Feeder cell-free monkey ES cell derivatives or clonally derived ES cells will be subjected to paradigms involving growth factor removal with additions of conditioned medium, extracts of monkey tissues, or precursors of the dopaminergic and cholinergic synthesis pathway. The efficiency of cell type differentiation will be determined by counting dopaminergic and cholinergic cells that will be identified by specific protein or gene markers. This is a revised application that is focused on understanding the directional differentiation of monkey ES cells in vitro into dopaminergic and cholinergic neurons for future transplantation studies with monkey models of Parkinson's or Alzheimer's diseases. Thesaurus Terms: biological signal transduction, cell communication molecule, cell differentiation, cell growth regulation, embryonic stem cell, neurogenesis choline acetyltransferase, fibroblast growth factor, laminin, membrane transport protein, neurotransmitter receptor, receptor expression, tyrosine 3 monooxygenase tissue /cell culture Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 30-SEP-2002 Project End: 31-AUG-2004 ICD: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE IRG: MDCN
Grant Number: 5R37HD019899-19 PI Name: CONN, PAUL M. PI Email: PI Title: PROFESSOR & CHAIRMAN OF PHARMACOLOGY Project Title: GONADOTROPIN-RELEASING HORMONE ACTION Abstract: Studies dealing with the biochemical and molecular mechanism of gonadotropin releasing hormone (GnRH) action have served to: (a) identify new clinical and veterinary uses for GnRH and its analogs, (b) bring these uses to fruition through a rational process based on understanding of the mechanism of hormone action, and (c) provide the means for anticipating, understanding, and ameliorating side-effects of the agents. FDA approvals of GnRH agonists for the treatment of prostate cancer, endometriosis, and precocious puberty, as well as for the use of natural sequence GnRH to induce ovulation, and to test the hypothalamic-pituitary-gonadal axis, are examples of the clinical usefulness derived from these fundamental observations. There are multiple advantages to the study of GnRH- stimulation of the gonadotrope that make it facile to collect interpretable data: (a) GnRH stimulation of the gonadotrope cell has clearly defined, specific and measurable endpoints: release of endocrine (and potentially endocrine) substances (LH, FSH, secretogranin II, alpha- subunit of gonadotropin), regulation of target cell sensitivity, regulation of the GnRH receptor, and biosynthesis of released substances. (b)The releasing hormone itself (as well as its agonists and antagonists) can be radioiodinated to high specific activity. (c) Many (>3,000) analogs exist that can be chemically derivatized without loss of biological activity. Antagonists and agonists which bind the receptor with greater affinities than the natural sequence GnRH are available. (d) Virtually all known agonists and antagonists are "pure" in action and metabolically stable agonists are available. The present project is divided into three areas of focus that are detailed in the "Specific Aims" section and form the basis of organization of the "Research Design and Methods" section. The first area will provide information on the structure of the GnRH receptor and early actions following the interaction of the receptor with GnRH and its analogs. At the present time, due to technical difficulties in dealing with the receptor itself, it has not been possible to use standard techniques to purify or even solubilize the receptor for a protracted period. Accordingly, these studies should advance our understanding of the receptor in the mechanism of GnRH action. The second area will provide information on the relationship between the multiple effector mechanisms already implicated in GnRH action with the multiple actions stimulated by the releasing hormone (release of multiple endocrine substances, regulation of target cell responsiveness, biosynthesis, and regulation of receptor number). This is important since considerable confusion remains regarding these relationships. The third area involves understanding the molecular sites of action of activin and inhibin in relation to GnRH action and are significant to understand the actions of these agent in vivo. Thesaurus Terms: biological signal transduction, gonadotropin releasing factor, hormone receptor, hormone regulation /control mechanism G protein, adenosine diphosphate, calcium, calcium channel, calmodulin, follicle stimulating hormone, inhibin, inositol phosphate, luteinizing hormone, molecular weight, peptide hormone analog, phosphorylation, protein kinase C, receptor binding, ribose phosphate affinity labeling, immunoprecipitation, laboratory rabbit, laboratory rat, radioimmunoassay, tissue /cell culture Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-APR-1984 Project End: 30-NOV-2003 ICD: NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT IRG: NSS
Grant Number: 5U01AG021382-02 PI Name: ZELINSKI-WOOTEN, MARY B. PI Email: zelinski@ohsu.edu PI Title: AFFILIATE ASSISTANT SCIENTIST Project Title: Ovarian Aspects of Caloric Restriction Abstract: DESCRIPTION (provided by applicant): Caloric restriction (CR) extends the life span, slows aging and retards age-related disease processes in short-lived mammalian species. Reproductive aging encompasses a life-long continuum of follicle depletion in the ovary that leads to decreased fecundity in older women and culminates in menopause, the cessation of ovarian/menstrual cyclicity, and its associated health-related risks. Caloric restriction delays the onset of ovarian follicular loss in rodents. Whether ovarian senescence is likewise suspended during CR in primates is not well understood. Using young and old female rhesus monkeys undergoing acute and long-term CR and their age-matched controls (CON), we propose to assess whether CR alters ovarian aging by determining: 1 ) the patterns and levels of gonadotropin and steroid hormones as well as inhibin-related proteins during spontaneous menstrual cycles and the peri-menopausal period; 2) the responsiveness of somatic cells of the ovarian follicle, i.e. granulosa cells, to exogenous gonadotropin or "fertility" treatment, and resultant follicular growth and maturation; and 3) gene expression in luteinizing granulosa cells and localization of protein factors involved in the pro- or anti-apoptotic (cell death) pathways in the ovarian follicle. Hormonal profiles will be measured in daily samples during 3 consecutive menstrual cycles in all animals, and frequent sampling will be done over a 6-hour period during the early follicular phase of 21 cycles to determine gonadotropin pulsatility in acute CON and CR animals. AII animals will receive recombinant human gonadotropins to stimulate the growth of multiple pre-ovulatory follicles followed by a bolus of hCG to induce peri-ovulatory events. Progesterone production by luteinizing granulosa cells in the presence or absence of hCG in vitro will be measured, and global gene expression will be assessed by microarray technology. Ovarian morphology and protein localization will be examined with histochemical and immunocytochemical analyses in acute CON and CR animals. These studies will provide valuable insight into the potential impact of CR on the mechanisms of ovarian aging in primates. Thesaurus Terms: aging, caloric dietary content, dietary restriction, ovary age difference, cell proliferation, chorionic gonadotropin, cooperative study, corpus luteum, gene expression, gonadotropin, graafian follicle, inhibin, menopause, menstrual cycle, protein localization, reproductive system pharmacology, steroid hormone Macaca mulatta, animal old age, histochemistry /cytochemistry, human genetic material tag, mature animal, microarray technology, nutrition related tag Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 30-SEP-2002 Project End: 31-AUG-2005 ICD: NATIONAL INSTITUTE ON AGING IRG: ZAG1
Grant Number: 5U24RR018107-02 PI Name: AXTHELM, MICHAEL K. PI Email: axthelmm@ohsu.edu PI Title: ASSOCIATE SCIENTIST Project Title: ESTABLISHMENT OF SPECIFIC PATHOGEN FREE RHESUS AND PIGTA Abstract: DESCRIPTION (Provided by applicant): Opportunistic human viruses and their simian counterparts are similar in their genetic makeup and induce a similar spectrum of diseases in their immunosuppressed hosts. Experimental lentivirus infections in rhesus macaques are widely recognized as the most import animal model for AIDS-related research and there is an urgent need to expand breeding programs to meet future AIDS vaccine and pathogenesis research program needs. Development of an Indian rhesus macaque breeding colony free of opportunistic viral agents is proposed to enhance the usefulness of this unique resource for studies focused on AIDS-related opportunistic infections. Housing space is proposed to provide a protected environment for this unique Expanded Specific Pathogen Free (ESPF) macaque resource and to facilitate enhanced macaque breeding efforts. Thesaurus Terms: Macaca mulatta, Macaca nemestrina, animal colony, germ free condition, opportunistic infection AIDS vaccine, Herpesviridae, Retroviridae, cooperative study, genetic strain, simian immunodeficiency virus, virus disease Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 30-SEP-2002 Project End: 31-AUG-2007 ICD: NATIONAL CENTER FOR RESEARCH RESOURCES IRG: RIRG
Grant Number: 5U42RR016025-04 PI Name: AXTHELM, MICHAEL K. PI Email: axthelmm@ohsu.edu PI Title: ASSOCIATE SCIENTIST Project Title: ESTABLISHMENT OF SPEC PATHOGEN FREE RHESUS MACAQUE COLON Abstract: The projected need for Indian rhesus macaques for AIDS-related research exceeds availability from current domestic breeding programs and there is an urgent need to expand breeding programs for Indian rhesus macaques for future AIDS vaccine and pathogenesis studies. Further, rhesus macaques with defined histocompatibility complex genotypes and known pedigree are becoming increasingly important for research to understand the biologic variation in the immune response. The long-term objective of this application is to expand the Oregon Regional Primate Center's specific pathogen-free Indian rhesus resource and sufficiently characterize their MHC haplotype to permit selected pedigree breeding for MHC class I alleles useful in AIDS research. The specific aims for accomplishing these objectives include intensively managing a subpopulation of the Center's SPF Indian rhesus macaque breeding colony to maximize production of genetically diverse females to expand the breeding capacity of the colony. Selective breeding of MHC-typed animals will be used to enhance production of future breeder males that are homozygous for the MAMU-A 01 allele, an important allele for assessing virus-specific cell-mediated immune function in simian immunodeficiency virus vaccine models for preventing AIDS virus infection. Breeder males that are homozygous for the MAMU-A 01 allele can be used to efficiently produce large numbers of offspring carrying the desired allele without inbreeding. The breeding colony will be initially typed for eight MHC alleles and managed for the production of MHC-defined offspring of known parentage. New sheltered field cage housing is proposed to protect the animals from infectious agents in the environment. Thesaurus Terms: Macaca mulatta, animal breeding, animal colony, germ free condition MHC class I antigen, animal care, epizootiology, genotype, major histocompatibility complex, simian immunodeficiency virus Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 30-SEP-2000 Project End: 31-AUG-2005 ICD: NATIONAL CENTER FOR RESEARCH RESOURCES IRG: ZRR1
Grant Number: 5U54HD018185-20 PI Name: STOUFFER, RICHARD L. PI Email: stouffri@ohsu.edu PI Title: SENIOR SCIENTIST AND HEAD Project Title: COOPERATIVE RESEARCH ON INFERTILITY IN PRIMATES Abstract: This study from the Oregon Regional Primate Research Center (ORPRC) establishes a Specialized Cooperative Center (U54) in Reproduction Research that addresses the causes and cures of human infertility disorders as they rate to neural, gonadal and gamete deficits. Each of the four research projects utilizes non-human primates in preclinical trials, and three contain clinical components in an effort to relate new information rapidly to the human situation. The first project, "Effect of Common Life Stresses on Fertility," will test the hypothesis that mild, relatively acute forms of stress (e.g., dieting and psychosocial changes) influence fertility in adult monkeys. Also, data will be generated to determine if success rates in treating women with tubal infertility correlate with patients' stress levels. The second project, "GABAergic Control of LHRH Neuronal Function," will utilize genetic and cellular manipulations to probe the role of GABAergic innervation to the development, migration and function of the LHRH neuronal network. Both Projects have components that focus on potential lesions in the LHRH system in the etiology of infertility. The third project, "Biomedical Basis of Intracytoplasmic Sperm Injection (ICSI," explores whether the molecular and cellular events following ICSI are comparable to natural fertilization. Information from these experiments will be used to test novel strategies for evaluation the quality of microinjected human sperm. The fourth project, "Vasoactive Factors in ART Cycles and Ovarian Hyperstimulation Syndrome (OHSS), " will test the hypothesis that methods used to generate multiple mature follicles/corpora lutea in ART cycles lead to an aberration in ovarian production of vascular endothelial-specific substances that could cause OHSS symptoms. Thus, Projects III and IV attempt to improve methods of diagnosing and treating infertility. These projects will be supported by three core (Assisted Reproductive Technologies or ART, Core A, Imaging & Morphology, Core B; Molecular & Cellular Biology, Core C) laboratories operating in an "open access" formula. The Administrative Core will foster intra- and inter-center cooperation to facilitate basic and clinical research. The U54 Center at ORPRC will be established within a strong interactive, collegial environment that has existed for two decades during prior NICHD Population Center support. Thesaurus Terms: cooperative study, reproduction Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 01-SEP-1984 Project End: 31-MAR-2004 ICD: NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT IRG: ZHD1
Grant Number: 7R01NS039550-04 PI Name: SHERMAN, LARRY S. PI Email: shermanl@ohsu.edu PI Title: ASSOCIATE PROFESSOR Project Title: ROLE OF CD44 IN PERIPHERAL NERVE DEVELOPMENT AND INJURY Abstract: The principle goal of the proposed studies is to elucidate the role of the CD44 family of transmembrane glycoproteins in perpheral nerves during development and following injury. Peripheral nerve development and maintenance require tightly regulated interactions between axons and Schwann cells. Schwann cell survival, proliferation and differentiation are influenced by axon-derived signals. One key signal is glial growth factor (GGF), which activates heterodimers of the receptor protein tyrosine kinases erbB2 and erbB3 in Schwann cells. GGF and its receptors have also been implicated in Wallerian degeneration, a process that occurs following nerve injury and which includes that induction of Schwann cell proliferation. The means by which GGF and related axon-derived signals promote erbB2-erbB3 heterodimerization and kinase activity are unclear. Our central hypothesis is that CD44 proteins are required for GGF signaling and in Schwann cell survival, proliferation and differentiation. CD44 has been implicated in cell-cell and cell- matrix interactions, and in growth factor presentation to high affinity cell surface receptors. Our preliminary data indicate the CD44 is essential for erbB2-erbB3 heterodimerization in Schwann cells, and that inhibition of CD44 expression results in Schwann cell apoptosis. We also found that CD44 is expressed in developing peripheral nerve at high levels when erbB2 expression is high and during active Schwann cell proliferation. We propose that CD44 acts by facilitating the interaction between axon- derived GGF and erbB receptors on the Schwann cell surface. We will test this notion experimentally with the following specific aims: (1) To ascertain whether CD44 acts as a low affinity GGF receptor; (2) To define the structural domains of CD44 that mediate interactions with ErbB2 and ErbB3 using cells expressing mutant CD44 proteins; (3) To determine if CD44 is required for Schwann cell survival, proliferation and/or differentiation during peripheral nerve development and Wallerian degeneration by comparing wild type mice and trasngenic mice whose Scwann cells lack CD44. Understanding how CD44 mediates the GGF-erbB2-erbB3 signaling complex will provide insight into the molecular mechanisms underlying normal peripheral nerve development, and may contribute significantly to our understanding of numerous conditions and diseases where axonal degeneration occurs, including nerve trauma, spinal cord injuries, and peripheral neuropathies. Thesaurus Terms: CD44 molecule, nerve injury, nervous system regeneration, neurogenesis, neurotrophic factor, protein tyrosine kinase Schwann cell, cell differentiation, cell proliferation, enzyme activity, growth factor receptor, intermolecular interaction, peripheral nervous system, wallerian degeneration laboratory mouse, transgenic animal Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 97239-3098 Fiscal Year: 2003 Department: NONE Project Start: 17-DEC-1999 Project End: 30-NOV-2004 ICD: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE IRG: ZRG1
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