Grant number: 1C06RR020178-01
PI Name:  DORSA, DANIEL
PI Email:  dorsad@ohsu.edu 
PI Title:  PROFESSOR
Project Title:  EXTRAMURAL RESEARCH FACILITIES CONSTRUCTION
$3,650,000

Abstract: DESCRIPTION (provided by applicant): Oregon Health & Science University (OHSU) seeks funds to construct Phase I of a new building at the Oregon National Primate Research Center (ONPRC). The proposed facility will be called the Non-SPF Group Housing and Central Support Building. Rapid research growth, expansion of Specific Pathogen Free (SPF) breeding colonies, and enforcement by USDA of the "Draft Policy on Environment Enhancement for Nonhuman Primates," necessitate additional social housing for nonhuman primates (NHPs) and increased animal central support space. Our long-term objectives are to: 1) provide housing for NHPs in social groups or pairs whenever possible; 2) protect growing populations of SPF Indian Origin Rhesus Macaques; 3) free-up cage space for the growing number of research projects; and 4) provide state-of-the-art central support facilities to benefit all NHPs at ONPRC. To accomplish these objectives, we are proposing to construct a Non-SPF Group Housing and Central Support Building in two phases, the first phase to be funded with this C06 grant. Phase 1 of the proposed building will be 12,690 gross square feet (gsf) and will include 16 animal holding rooms to house up to 160 adult non-SPF NHPs in groups of 8-10 animals, a non-SPF clinic/hospital/nursery, a new central diet kitchen with pantry, walk-in cooler, walk-in freezer, centralized commercial feed storage, and a separate room to prepare and distribute produce for NHP enrichment as part of the Psychological Well Being program. Specific Aim 1 proposes to construct indoor group housing and support facilities (clinic, hospital, nursery) for non-SPF NHPs. Specific Aim 2 proposes to construct central nutritional and enrichment support facilities (diet kitchen, pantry, cooler, freezer, produce processing, and feed storage) to service all OHSU NHPs.

Thesaurus Terms: Primate, biomedical facility, building /facility design /renovation, housing
animal care

Institution:  OREGON HEALTH & SCIENCE UNIVERSITY
 PORTLAND, OR 972393098
Fiscal Year:  2004
Department:  NONE
Project Start:  20-SEP-2004
Project End:  19-SEP-2006
ICD:  NATIONAL CENTER FOR RESEARCH RESOURCES
IRG:  STRB

Grant number: 1R01HD042710-01A2
PI Name:  JENSEN, JEFFREY T.
PI Email:  jensenje@ohsu.edu 
PI Title:  
Project Title:  PDE3 Inhibitors: Selective Blockers of Oocyte Maturation
$321,975

Abstract: DESCRIPTION (provided by applicant): The need for contraception has never been greater, as evidenced by a world population of over 6 billion, and greater than one billion young people currently in the prime reproductive years. Overpopulation is a pernicious public health affliction that ravages infrastructure and limits the achievement of hard fought gains in overall health status by threatening the basic infrastructure of humanity. Contraception also provides significant preventative health benefits by reducing unintended pregnancy, abortions, and unwanted births. As side effects, access, and cultural or moral objections limit the universal acceptance of current methods, research into novel contraceptives is warranted. The purpose of this grant is to explore the hypotheses that selective expression of phosphodiesterase (POE) isoforms exists in the primate ovary, that selective blockade of PDE3 can be exploited to prevent spontaneous and gonadotropin induced oocyte maturation, and that chronic treatment with a PDE3 inhibitor can prevent pregnancy in primates without affecting menstrual cyclicity or luteal function. The Specific Aims are to: (1) Describe the basic biology of PDE expression in the primate ovary. The goal of this aim is to characterize the PDE isoenzymes expressed in the macaque ovary. An additional goal will be to test novel PDE3 inhibitors for their ability to prevent spontaneous resumption of meiosis in vitro to learn if greater selectivity or potency can be achieved. (2) Determine if PDE3 inhibitors prevent oocyte maturation, but not ovulation and function of the corpus luteum in rhesus monkeys undergoing controlled ovarian stimulation (COS) protocols or during natural cycles in vivo. The goals of this aim will be to document that oocyte inhibition can be effectively and selectively achieved in vivo, and to investigate the systemic toxicity of PDE3 inhibitors at pharmacologically active dosages in vivo. 3) Determine whether PDE3 inhibitors function as contraceptive agents in regularly cycling rhesus monkeys in paired mating situations. The goal of this aim is to document the efficacy and practicality of systemic administration of a PDE3 inhibitor as a potential contraceptive through longer-term observation. Experimental designs will include: characterization of PDE gene products (mRNA and protein) in somatic and germ cells from ovarian tissue (Aim 1), in vitro incubation of immature oocytes and granulosa cells with PDE3 inhibitors (Aim 1), in vivo administration of a PDE3 inhibitor to monkeys during COS and spontaneous menstrual cycles, followed by follicular aspiration to assess oocyte maturation and fertilizability in vitro, plus endocrine and toxicity measurements (Aim 2), and chronic administration of a PDE 3 inhibitor to fertile females caged with fertile males to assess contraceptive efficacy and long term toxicity (Aim 3). This approach is expected to provide a foundation for future Phase 1 trials of PDE3 inhibitors in humans.

Thesaurus Terms: 3'5' cyclic nucleotide phosphodiesterase, contraceptive, drug discovery /isolation, drug screening /evaluation, egg /ovum, oogenesis, phosphodiesterase inhibitor, protein isoform
corpus luteum, fertilization, gene expression, isozyme, meiosis, menstrual cycle, ovulation, pharmacokinetics, protein localization
Macaca mulatta, in situ hybridization, laparoscopy

Institution:  OREGON HEALTH & SCIENCE UNIVERSITY
 PORTLAND, OR 972393098
Fiscal Year:  2004
Department:  OBSTETRICS AND GYNECOLOGY
Project Start:  15-MAY-2004
Project End:  29-FEB-2008
ICD:  NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
IRG:  REB

Grant number: 1R03AG023280-01
PI Name:  SHERMAN, LARRY S.
PI Email:  shermanl@ohsu.edu 
PI Title:  ASSOCIATE PROFESSOR
Project Title:  Hyaluronate in aging-related gliosis and demyelination
$71,550

Abstract: The goal of this pilot study is to understand the contribution of the glycosaminoglycan hyaluronate (HA) to demyelination and gliosis in the aging central nervous system (CNS). Age-related changes in cortical white matter have been implicated in cognitive impairment. These changes include alterations in astrocytes that mimic reactive gliosis and the breakdown of myelin sheaths. In neurodegenerative conditions, glial cells overexpress the CD44 transmembrane glycoprotein, a receptor for HA. We recently found that chronic CD44 overexpression by oligodendrocytes results in increased HA and progressive demyelinating disease, while alterations in the HA-based extracellular matrix promote reactive astrogliosis. Our preliminary data suggest that many of the changes in CD44 expression and HA distribution that occur in neurodegenerative conditions also occur in the aging brain. These findings support the hypothesis that CD44 and HA accumulation are induced during the course of normal aging and may contribute to aging-related demyelination and astrogliosis. Here, we will test this hypothesis by (1) examining how HA accumulates in the aging primate and rodent CNS; and (2) testing the requirement for CD44 in promoting gliosis and related cognitive changes in the aging rodent brain. These studies will provide substantial insight into the contribution of CD44 and HA to alterations in elderly white matter, and will provide substantial preliminary data for a larger, future proposal aimed at determining how targeting HA and CD44 can reverse altered glial cell function and, ultimately, cognitive impairment.

Thesaurus Terms: aging, glia, gliosis, hyaluronate, myelinopathy
CD44 molecule, astrocyte, central nervous system, extracellular matrix, gene expression, glycoprotein, mucopolysaccharide, neural degeneration, white matter
chromatography, confocal scanning microscopy, immunocytochemistry, in situ hybridization, laboratory mouse, polymerase chain reaction

Institution:  OREGON HEALTH & SCIENCE UNIVERSITY
 PORTLAND, OR 972393098
Fiscal Year:  2004
Department:  CELL AND DEVELOPMENTAL BIOLOGY
Project Start:  01-APR-2004
Project End:  31-MAR-2006
ICD:  NATIONAL INSTITUTE ON AGING
IRG:  NIA

Grant number: 1S10RR019434-01
PI Name:  CORNEA, ANDA
PI Email:  corneaan@ohsu.edu 
PI Title:  
Project Title:  Stereology & Live Cell Imaging Workstation
$214,760

Abstract: DESCRIPTION (provided by applicant): This application requests funds for purchasing an imaging workstation that enables stereological measurements based on multichannel fluorescence identification of features to be counted, time-lapse live cell fluorescence imaging and measuring Fluorescence Resonance Energy Transfer (FRET) between cyan and yellow fluorescent proteins (CFP and YFP). Morphometric measurements, such as numbers and sizes of cells, density of neuronal fibers in brain circuits, density of synapses in specific areas or on specific cells, number and stages of ovarian follicles are important determinants of physiological changes associated with conditions that constitute the focus of research of multiple NIH-funded grants at the Oregon National Primate Research Center (ONPRC). Estimating these parameters based on two-dimensional microscopic images is very laborious and is affected by biases, if sampling and analysis techniques of modern stereology are not followed. A stereology compatible imaging system is therefore necessary for the research of our investigators. For other projects, use of green fluorescent proteins and their mutants has created opportunities for identifying the intracellular location, translocation and function of genes of interest. A particularly attractive application is the visualization and measurement of the interaction between biomolecules in physiologically relevant states, in real time, using FRET, especially between CFP and YFP. This is not possible with the instruments currently available to us, but necessary for our NIH-funded investigators. This workstation will be part of the Imaging and Morphology Core of ONPRC located on the West campus of the Oregon Health & Sciences University and will enable the core to provide services that are either not possible now, or that will be significantly improved in accuracy and speed. An internal advisory committee will establish procedures and oversee the general operation of the imaging workstation. The principal investigator will be in charge of the installation, training, scheduling, use and maintenance of the new imaging workstation. The institution is committed to maintenance of instrument and providing oversight, support for staff salary and training.

Thesaurus Terms: biomedical equipment purchase, cell morphology, imaging /visualization /scanning
fluorescence microscopy, fluorescence resonance energy transfer, fluorescent dye /probe, time resolved data
bioimaging /biomedical imaging

Institution:  OREGON HEALTH & SCIENCE UNIVERSITY
 PORTLAND, OR 972393098
Fiscal Year:  2004
Department:  NONE
Project Start:  01-APR-2004
Project End:  31-MAR-2005
ICD:  NATIONAL CENTER FOR RESEARCH RESOURCES
IRG:  ZRG1

Grant number: 2P51RR000163-45  (base grant)
PI Name:  KOHLER, PETER O.
PI Email:  
PI Title:  PRESIDENT
Project Title:  SUPPORT FOR NATIONAL PRIMATE RESEARCH CENTER
$11,065,887

Abstract: DESCRIPTION (provided by applicant): The 5-year renewal for Grant P51 RR00163-45 requests funding for programs that support biomedical research at the ONPRC. The application reflects a significant expansion in animal resources and research service cores that is required to support the growth in research programs (external grant funding has doubled during the past 5 years) and to greatly expand the Specific Pathogen Free (SPF) Indian-derived Rhesus Breeding Colony. The aims of this application are to: 1) provide the infrastructure support necessary to conduct advanced biomedical research for which nonhuman primates (NHPs) are used to accelerate progress in understanding human diseases, leading to better health; 2) enhance the Center's resources (scientific expertise, laboratories, animal services, research service cores and equipment) so as to serve as a regional, national and international resource; 3) increase the availability of defined NHPs for biomedical research through support of our programs to expand the SPF Indian-derived Rhesus macaque breeding program that produces animals free of viruses such as herpes B and with defined parentage and genotype; and 4) expand research programs utilizing NHPs in genetics, infectious diseases and vaccine development, aging, neuroscience and women's health. To accomplish these aims, support is requested in four broad areas. Administration and Center Operations provide the administrative and service support that is required for all aspects of the ONPRC. Units include the Director's Office, Business Services, Facilities and Property, and Information Technology and Engineering. Research Infrastructure encompasses the service units that provide the most technologically advanced research support, such as Animal Resources (Business and Research Coordination, Clinical Medicine, NHP Resources, Operations, Pathology Services, Psychological Well Being and Surgery Services), Research Service Cores (Assisted Reproductive Technology, Electrophysiology, Endocrine Services, Flow Cytometry, Imaging and Morphology, Molecular and Cellular Biology, Monoclonal Antibody, Research Library, and Virology) and Special Resources Programs (Alternative NHP Species, Genetics Resource and Informatics, Aging NHPs and Rhesus Breeding). Scientific Resources includes support for the three research divisions that house ONPRC scientists: the Divisions of Pathobiology and Immunology, Neuroscience, and Reproductive Sciences. Funding is also requested for 4 resource-related research projects and the pilot project program. Improvement and Modernization includes funds for equipment and alterations and renovations. The funds requested will provide support for the research infrastructure that is required to conduct the most advanced biomedical research using NHPs as models of human health and disease, and to meet the needs of new emerging areas of research in which the NHP model is critical, such as functional genomics and biodefense initiatives.

Thesaurus Terms: Primate, biomedical facility

Institution:  OREGON HEALTH & SCIENCE UNIVERSITY
 PORTLAND, OR 972393098
Fiscal Year:  2004
Department:  NONE
Project Start:  01-MAY-1997
Project End:  30-APR-2009
ICD:  NATIONAL CENTER FOR RESEARCH RESOURCES
IRG:  ZRR1

Grant number: 2R01HD019899-20
PI Name:  CONN, P. MICHAEL.
PI Email:  connm@ohsu.edu 
PI Title:  PROFESSOR & CHAIRMAN OF PHARMACOLOGY
Project Title:  Gonadotropin-Releasing Hormone Action
$531,117

Abstract: DESCRIPTION (provided by applicant): This is a request for continuation of a program begun in 1979. Since isolation of gonadotropin releasing hormone (GnRH) in the early 1970s, basic and clinical discoveries have presented uses for GnRH analogs. Among these are treatments for endometriosis/fibroids, polycystic ovary disease, perimenopause, protection of the ovary and testes (i.e., during chemotherapy), assisted reproduction, precocious puberty, male and female birth control, prostatic, ovarian and mammary carcinomas, cryptorchidism, and other conditions. The first GnRH agonist was approved for use in the US in 1985 (Leuprolide (Lupron, TAP/Abbott)); Buserelin (Hoechst) was already in use in Europe at that time. Each of these molecules, and other agonists, continue to be available for international use. Due to patent expirations, they are now sold by various companies and in an array of formulations and delivery systems, adding to their potential utility and comprising a world-wide market in excess of $1.5 billion. Studies of GnRH action have provided a great deal of useful, basic information to the field of neuroendocrine peptides. Among the faciliatory features of GnRH and its analogs are ease of preparing GnRH analog radioligands, availability of model systems with measurable endpoints, the relative specificity of actions, size (i.e., TRH is very small and chemical modifications frequently destroy binding activity, while CRF and GHRH are large and analogs are relatively difficult to synthesize; GnRH is virtually impossible to denature), and availability of vectors encoding the GnRH receptor and many mutants (along with fluorescent probes) from a range of species and tissues. In addition, nearly 10,000 biologically active GnRH analogs are now described in the scientific and patent literature, providing an exquisite database for understanding structure-activity relations; these analogs include well-defined full agonists and full antagonists, many of which are metabolically stable or have other desirable characteristics. Improved understanding of the mechanism of action of this hormone will likely lead to opportunities for improved drugs (i.e., fewer side effects, orally active and cheaper) and more specific therapies. "Non-traditional" approaches also have potential; approaches involving some of these are described in the present proposal. The specific aims of this study will advance both our basic understanding and will identify new sites and approaches potentially amenable to therapeutic intervention.

Thesaurus Terms: gonadotropin releasing factor, hormone regulation /control mechanism, pharmacokinetics
G protein, glycosylation, guanosinetriphosphatase activating protein, hormone receptor, mathematical model, model design /development, protein localization, protein protein interaction, protein structure function, receptor coupling, stimulant /agonist, stoichiometry
animal tissue, tissue /cell culture

Institution:  OREGON HEALTH & SCIENCE UNIVERSITY
 PORTLAND, OR 972393098
Fiscal Year:  2004
Department:  PHYSIOLOGY AND PHARMACOLOGY
Project Start:  01-APR-1984
Project End:  28-FEB-2009
ICD:  NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
IRG:  BCE

Grant number: 2U54HD018185-21
PI Name:  STOUFFER, RICHARD L.
PI Email:  stouffri@ohsu.edu 
PI Title:  SENIOR SCIENTIST AND HEAD
Project Title:  Cooperative Research on Infertility in Primates
$1,430,056

Abstract: DESCRIPTION (provided by applicant): The Oregon National Primate Research Center (ONPRC) proposes to continue a Specialized Cooperative (U54) Center in Reproduction Research that addresses the causes and cures of human fertility disorders as related to neural, gonadal and gamete deficits. Each research project utilizes female nonhuman primates in translational research, and two contain clinical components in an effort to relate new information to improving women's health. Project I, "Effect of Common Life Stresses on Fertility," expands investigations on the neuroendocrine and gonadal lesions associated with a combination of mild, acute forms of stress (e.g., dieting and psychosocial changes) and whether treatment to increase serotonergic tone ameliorates these lesions, as well as associated infertility. Project II, "Novel Mechanisms Underlying the Transsynaptic Control of LHRH Release," continues genetic and cellular manipulations to evaluate the role of GABA input, as well as that of other novel genes (FXYD, Nell2, and lAP-1) in upstream neuronal pathways, in controlling the function of LHRH neurons and normal reproductive cyclicity. Project III, "Angiogenic and Angiolytic Factors in Natural and Controlled Ovarian Stimulation (COS) Cycles," will examine the expression and action of angiopoietin (Ang) and endocrine gland (EG)-VEGF in the preovulatory follicle and corpus luteum, and whether alterations in Ang, EG-VEGF, as well as VEGF-A, cause ovarian disorders, such as ovarian hyperstimulation syndrome (OHSS). Project IV, "Proteomic Profiling: Markers of Normal and Abnormal Follicle Development," is a pilot project to determine the feasibility of using proteomic analyses of follicular fluid to identify novel proteins involved in follicular development and ovulation, as well as markers or mechanisms in polycystic ovarian syndrome (PCOS). Thus, all projects attempt to relate basic reproductive processes to the etiology and/or treatment of infertility. These projects will be supported by three core laboratories (Assisted Reproductive Technologies or ART, Core A; Imaging & Morphology or IM, Core B; Molecular & Cellular Biology or MCB, Core C) operating in an "open access" formula. The Administrative Core will foster intra- and inter-center cooperation in basic and applied research. The ONPRC U54 Center is flourishing through a strong scientific environment at the Primate Center, collaborations in women's health at the Oregon Health & Science University, and interactions with other SCCPRR programs.

Thesaurus Terms: cooperative study, fertility
Macaca mulatta, clinical research, human subject

Institution:  OREGON HEALTH & SCIENCE UNIVERSITY
 PORTLAND, OR 972393098
Fiscal Year:  2004
Department:  PHYSIOLOGY AND PHARMACOLOGY
Project Start:  01-APR-1997
Project End:  31-MAR-2009
ICD:  NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
IRG:  ZHD1

Grant number: 3R01DK060685-02S1
PI Name:  GROVE, KEVIN L.
PI Email:  grovek@ohsu.edu 
PI Title:  
Project Title:  NPY Feeding Circuits during Development
$500,014

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 972393098
Fiscal Year:  2004
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: 3R01DK065900-02S1
PI Name:  SIMERLY, RICHARD B.
PI Email:  simerlyr@ohsu.edu 
PI Title:  SENIOR SCIENTIST
Project Title:  Development of Leptin-Sensitive Hypothalamic Pathways
$522,684

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 972393098
Fiscal Year:  2004
Department:  CELL AND DEVELOPMENTAL BIOLOGY
Project Start:  30-SEP-2003
Project End:  31-JUL-2006
ICD:  NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
IRG:  END

Grant number: 3R01NS039550-04S1
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
$24,944

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 972393098
Fiscal Year:  2004
Department:  CELL AND DEVELOPMENTAL BIOLOGY
Project Start:  17-DEC-1999
Project End:  30-NOV-2004
ICD:  NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
IRG:  ZRG1

Grant number: 5D43TW000668-10
PI Name:  CONN, P. MICHAEL.
PI Email:  connm@ohsu.edu 
PI Title:  PROFESSOR & CHAIRMAN OF PHARMACOLOGY
Project Title:  INTERNATIONAL TRAINING AND RES. IN POPULATION AND HEALTH
$240,960

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 972393098
Fiscal Year:  2004
Department:  PHYSIOLOGY AND PHARMACOLOGY
Project Start:  30-SEP-1995
Project End:  30-APR-2006
ICD:  FOGARTY INTERNATIONAL CENTER
IRG:  ZHD1

Grant number: 5R01AG019914-04
PI Name:  URBANSKI, HENRYK F.
PI Email:  urbanski@ohsu.edu 
PI Title:  ASSOCIATE SCIENTIST
Project Title:  Effect of Aging and Caloric Restriction on Circadian Ph*
$259,289

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 972393098
Fiscal Year:  2004
Department:  PHYSIOLOGY AND PHARMACOLOGY
Project Start:  01-SEP-2001
Project End:  31-AUG-2005
ICD:  NATIONAL INSTITUTE ON AGING
IRG:  ZAG1

Grant number: 5R01AI042490-07
PI Name:  GRAVETT, MICHAEL G.
PI Email:  gravettm@ohsu.edu 
PI Title:  CHIEF
Project Title:  Experimental Model for Chorioamnionitis and Prematurity
$278,250

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 972393098
Fiscal Year:  2004
Department:  OBSTETRICS AND GYNECOLOGY
Project Start:  01-SEP-1997
Project End:  31-MAY-2006
ICD:  NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
IRG:  HED

Grant number: 5R01CA075922-08
PI Name:  WONG, SCOTT W.
PI Email:  wongs@ohsu.edu 
PI Title:  ASSOCIATE SCIENTIST
Project Title:  RHESUS HHV 8 HOMOLOGUE IN AIDS RELATED MALIGNACIES
$716,522

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 972393098
Fiscal Year:  2004
Department:  VACCINE AND GENE THERAPY INSTITUTE
Project Start:  01-JUL-1997
Project End:  30-JUN-2005
ICD:  NATIONAL CANCER INSTITUTE
IRG:  ZRG1

Grant number: 5R01DK060685-02
PI Name:  GROVE, KEVIN L.
PI Email:  grovek@ohsu.edu 
PI Title:  
Project Title:  NPY Feeding Circuits during Development
$225,601

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 972393098
Fiscal Year:  2004
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: 5R01DK065900-02
PI Name:  SIMERLY, RICHARD B.
PI Email:  simerlyr@ohsu.edu 
PI Title:  SENIOR SCIENTIST
Project Title:  Development of Leptin-Sensitive Hypothalamic Pathways
$341,850

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, genetically modified animal, histochemistry /cytochemistry, laboratory mouse, newborn animal, nutrition related tag, radiotracer

Institution:  OREGON HEALTH & SCIENCE UNIVERSITY
 PORTLAND, OR 972393098
Fiscal Year:  2004
Department:  CELL AND DEVELOPMENTAL BIOLOGY
Project Start:  30-SEP-2003
Project End:  31-JUL-2006
ICD:  NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
IRG:  END

Grant number: 5R01HD014643-22
PI Name:  SMITH, M SUSAN.
PI Email:  smithsu@ohsu.edu 
PI Title:  ASSOCIATE PROFESSOR OF PHYSIOLOGY
Project Title:  CONTROL OF GONADOTROPIN SECRETION DURING LACTATION
$248,102

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 972393098
Fiscal Year:  2004
Department:  PHYSIOLOGY AND PHARMACOLOGY
Project Start:  01-DEC-1979
Project End:  28-FEB-2006
ICD:  NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
IRG:  REN

Grant number: 5R01HD019182-19
PI Name:  BRENNER, ROBERT M.
PI Email:  brennerr@ohsu.edu 
PI Title:  SENIOR SCIENTIST
Project Title:  HORMONAL CONTROL OF THE FEMALE REPRODUCTIVE TRACT
$321,975

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 972393098
Fiscal Year:  2004
Department:  CELL AND DEVELOPMENTAL BIOLOGY
Project Start:  01-SEP-1984
Project End:  30-APR-2005
ICD:  NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
IRG:  REB

Grant number: 5R01HD020869-18
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
$357,750

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 972393098
Fiscal Year:  2004
Department:  PHYSIOLOGY AND PHARMACOLOGY
Project Start:  01-JUL-1985
Project End:  28-FEB-2006
ICD:  NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
IRG:  REB

Grant number: 5R01HD024870-17
PI Name:  OJEDA, SERGIO R.
PI Email:  ojedas@ohsu.edu 
PI Title:  SCIENTIST/DIVISION HEAD
Project Title:  NEURAL CONTROL OF THE PREPUBERTAL OVARY
$402,324

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 972393098
Fiscal Year:  2004
Department:  PHYSIOLOGY AND PHARMACOLOGY
Project Start:  01-FEB-1988
Project End:  31-JAN-2006
ICD:  NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
IRG:  REB

Grant number: 5R01HD025123-15
PI Name:  OJEDA, SERGIO R.
PI Email:  ojedas@ohsu.edu 
PI Title:  SCIENTIST/DIVISION HEAD
Project Title:  NEUROENDOCRINOLOGY OF PUBERTY AND SEXUAL DEVELOPMENT
$382,727

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, genetically modified animal, laboratory mouse, laboratory rat

Institution:  OREGON HEALTH & SCIENCE UNIVERSITY
 PORTLAND, OR 972393098
Fiscal Year:  2004
Department:  PHYSIOLOGY AND PHARMACOLOGY
Project Start:  01-SEP-1990
Project End:  31-MAY-2005
ICD:  NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
IRG:  BCE

Grant number: 5R01HD029186-07
PI Name:  URBANSKI, HENRYK F.
PI Email:  urbanski@ohsu.edu 
PI Title:  ASSOCIATE SCIENTIST
Project Title:  Postnatal Development of Ionotropic Glutamate Receptors
$250,425

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 972393098
Fiscal Year:  2004
Department:  PHYSIOLOGY AND PHARMACOLOGY
Project Start:  01-SEP-1993
Project End:  31-MAR-2007
ICD:  NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
IRG:  BCE

Grant number: 5R01HD042000-03
PI Name:  HENNEBOLD, JON D.
PI Email:  henneboj@ohsu.edu 
PI Title:  
Project Title:  Epoxyeicosatrienoic Acids in the Ovulatory Follicle
$321,975

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, 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 972393098
Fiscal Year:  2004
Department:  OBSTETRICS AND GYNECOLOGY
Project Start:  01-JUN-2002
Project End:  31-MAY-2007
ICD:  NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
IRG:  REN

Grant number: 5R01HD043209-03
PI Name:  BRENNER, ROBERT M.
PI Email:  brennerr@ohsu.edu 
PI Title:  SENIOR SCIENTIST
Project Title:  Suppression of Breakthrough Bleeding in LNG-IUS Users
$304,575

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 972393098
Fiscal Year:  2004
Department:  CELL AND DEVELOPMENTAL BIOLOGY
Project Start:  27-SEP-2002
Project End:  30-JUN-2007
ICD:  NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
IRG:  ZHD1

Grant number: 5R01HL066118-03
PI Name:  SPINDEL, ELIOT R.
PI Email:  spindele@ohsu.edu 
PI Title:  SENIOR SCIENTIST
Project Title:  Nicotine & Alpha7 nicotinic receptor in lung development
$357,750

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, genetically modified animal, laboratory mouse, morphometry, plethysmography, tissue /cell culture

Institution:  OREGON HEALTH & SCIENCE UNIVERSITY
 PORTLAND, OR 972393098
Fiscal Year:  2004
Department:  CELL AND DEVELOPMENTAL BIOLOGY
Project Start:  01-JUL-2002
Project End:  30-JUN-2007
ICD:  NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
IRG:  HED

Grant number: 5R01MH062677-04
PI Name:  BETHEA, CYNTHIA L.
PI Email:  betheac@ohsu.edu 
PI Title:  SENIOR SCIENTIST
Project Title:  OVARIAN STEROID REGULATION OF SEROTONIN IN PRIMATES
$318,000

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 972393098
Fiscal Year:  2004
Department:  PHYSIOLOGY AND PHARMACOLOGY
Project Start:  01-FEB-2001
Project End:  30-NOV-2005
ICD:  NATIONAL INSTITUTE OF MENTAL HEALTH
IRG:  ZRG1

Grant number: 5R01MH065438-02
PI Name:  OJEDA, SERGIO R.
PI Email:  ojedas@ohsu.edu 
PI Title:  SCIENTIST/DIVISION HEAD
Project Title:  Molecular Specifiers of Neural Cell Plasticity
$350,300

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 972393098
Fiscal Year:  2004
Department:  PHYSIOLOGY AND PHARMACOLOGY
Project Start:  01-DEC-2002
Project End:  30-NOV-2006
ICD:  NATIONAL INSTITUTE OF MENTAL HEALTH
IRG:  BCE

Grant number: 5R01NS044330-03
PI Name:  WOLF, DON P.
PI Email:  wolfd@ohsu.edu 
PI Title:  SENIOR SCIENTIST
Project Title:  Genetically Modified Rhesus Monkeys
$618,891

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, genetically modified animal, model design /development, neurogenetics
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 972393098
Fiscal Year:  2004
Department:  OBSTETRICS AND GYNECOLOGY
Project Start:  15-AUG-2002
Project End:  30-JUN-2007
ICD:  NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
IRG:  ZRG1

Grant number: 5R01RR016030-04
PI Name:  WOLF, DON P.
PI Email:  wolfd@ohsu.edu 
PI Title:  SENIOR SCIENTIST
Project Title:  PROPAGATION OF MONKEY MODELS OF HUMAN DISEASE
$424,315

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 972393098
Fiscal Year:  2004
Department:  OBSTETRICS AND GYNECOLOGY
Project Start:  01-JUL-2001
Project End:  30-JUN-2006
ICD:  NATIONAL CENTER FOR RESEARCH RESOURCES
IRG:  RIRG

Grant number: 5R21MH063137-02
PI Name:  MACHIDA, CURTIS A.
PI Email:  machidac@ohsu.edu 
PI Title:  
Project Title:  Adrenergic Receptor Mechanisms in Antidepressant Therapy
$159,000

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 972393098
Fiscal Year:  2004
Department:  NONE
Project Start:  01-DEC-2002
Project End:  30-NOV-2005
ICD:  NATIONAL INSTITUTE OF MENTAL HEALTH
IRG:  ZRG1

Grant number: 5U01AG021382-03
PI Name:  ZELINSKI-WOOTEN, MARY B.
PI Email:  zelinski@ohsu.edu 
PI Title:  AFFILIATE ASSISTANT SCIENTIST
Project Title:  Ovarian Aspects of Caloric Restriction
$241,966

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 972393098
Fiscal Year:  2004
Department:  PHYSIOLOGY AND PHARMACOLOGY
Project Start:  30-SEP-2002
Project End:  31-AUG-2005
ICD:  NATIONAL INSTITUTE ON AGING
IRG:  ZAG1

Grant number: 5U24RR018107-03
PI Name:  AXTHELM, MICHAEL K.
PI Email:  axthelmm@ohsu.edu 
PI Title:  ASSOCIATE SCIENTIST
Project Title:  ESTABLISHMENT OF SPECIFIC PATHOGEN FREE RHESUS AND PIGTA
$1,252,364

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 972393098
Fiscal Year:  2004
Department:  NONE
Project Start:  30-SEP-2002
Project End:  31-AUG-2007
ICD:  NATIONAL CENTER FOR RESEARCH RESOURCES
IRG:  RIRG

Grant number: 5U42RR016025-05
PI Name:  AXTHELM, MICHAEL K.
PI Email:  axthelmm@ohsu.edu 
PI Title:  ASSOCIATE SCIENTIST
Project Title:  ESTABLISHMENT OF SPEC PATHOGEN FREE RHESUS MACAQUE COLON
$1,129,775

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 972393098
Fiscal Year:  2004
Department:  NONE
Project Start:  30-SEP-2000
Project End:  31-AUG-2005
ICD:  NATIONAL CENTER FOR RESEARCH RESOURCES
IRG:  ZRR1