Abstract: The overall goal of this project is to determine the roles that neuropeptide Y and POMC neuronal systems play regulating food intake in primate species. Neuropeptide Yis a very potent orexigenic agent in rodents, and POMC neurons that produce a-MSH appear to be in a position to play a key role in the inhibition of food intake in rodents, by acting through melanocortin type 4 (MC4) receptors. An interesting link between these systems has recently been established by our studies showing that in the rat arcuate nucleus NPY neurons contain mRNA for agouti-related peptide, AGRP, which is an endogenous inhibitor of the MC4 receptor. In contrast to the large number of studies seeking central neural systems that control food intake in rodents species, very little work has been done in primate species to determine if the control of food intake in the primate is similar to that in the rat. We have recently shown that administration of NPY can stimulate food intake in the adult male rhesus monkey, but that the feeding sensitive NPY system in the primate is much more complex than in the rat. The goal of the project we propose in this grant is to continue our studies in the non-human primate to determine (a) the degree to which food intake is controlled by NPY and POMC systems, (b) which population(s) of NPY and POMC neurons modulate food intake in the primate, and (c) what metabolic and neural signals regulate NPY and POMC lean primates. Increased understanding of the neural systems controlling food intake in the primate will provide important new information that will e useful in devising new treatments for obesity, a condition that leads to a large increase in human morbidity and mortality, resulting from an increased incidence of cardiovascular disease, stroke, diabetes, and associated problems in vision, kidney and liver function. Converse, this information will also be of utility in designing new treatments for people with life-threatening eating disorders, including anorexia and bulimia nervosa.

Thesaurus Terms:
hormone regulation /control mechanism, neuropeptide Y, nutrient intake activity, proopiomelanocortin brain mapping, hormone receptor, obesity Macaca mulatta, behavioral /social science research tag, nutrition related tag

Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 972393098
Fiscal Year: 2002
Department:
Project Start:
Project End:
ICD: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
IRG:

Abstract: Interleukin-8 (IL-8) is a cytokine that induces selective neutrophil chemotaxis and activation. Progesterone inhibits and antiprogestins stimulate production of IL-8 by chorion and decidua. Cervical ripening is associated with an influx of inflammatory cells into the cervix but the role of IL-8 in primate parturition remains to be established. To investigate the role of IL-8 in cervical ripening, we administered purified, human fibroblast-derived IL-8 intravaginally in 3 pregnant monkeys beginning at 137-9 days of gestation (term = 167 days) at a dose of 150 ng/kg. The cervix was evaluated by a modified Bishop=s Score. IL-8 was administered in vehicle (2.5% HPC with 300 ?l Evans Blue solution) by inserting a 1.0 or 3.0 ml syringe into the vaginal vault. Amniocentesis was performed at intervals to determine changes in amniotic fluid prostaglandins (PGs) and cytokines. After a recovery period of 4-5 days, a second dose of IL-8 (300 ng/kg) was administered. Res ults The effect of graded doses of intravaginal IL-8 on Modified Bishop=s Scores (mean ? SEM) is summarized in the table below. Two out of three animals showed an increase in amniotic fluid IL-8 concentrations but there were no changes in amniotic fluid PGE2 or PGF2?. One animal delivered after the second dose of IL-8. Dose 0 hours 24 hours 48 hours 72 hours 150 ng/kg 0.6 ? 0.3 5.6 ? 0.3* 5.0 ? 1.2* 5.3 ? 0.3* 300 ng/kg 2.0 ? 1.0 4.0 ? 3.0 6.5 ? 0.5* *different from time 0 hrs at P<0.05 (t-test). Conclusion Intravaginal application of purified human fibroblast-derived IL-8 induces significant cervical ripening in late pregnant rhesus monkeys. FUNDING NIH HD06159, Toray Industries Inc PUBLICATIONS Haluska GJ, Naruto M, Ida N, Cook MJ, Novy MJ. Induction of cervical ripening with interleukin-8 (IL-8) in pregnant rhesus monkeys. In Society for Gynecological Investigation Program and Abstracts 45th Annual Meeting (held in Atlanta, GA, March 11-14, 1998), p 190A (abstract #T615).

Thesaurus Terms:
endocrine gland /system, health care, mother /infant health care, reproductive system, women's health Mammalia, clinical research

Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 972393098
Fiscal Year: 2000
Department:
Project Start: 01-MAY-1978
Project End: 30-APR-2004
ICD: NATIONAL CENTER FOR RESEARCH RESOURCES
IRG:

Abstract: Fertilin, originally named PH-30, is a heterodimeric cell surface protein identified on guinea pig, rabbit and human spermatozoa. The alpha subunit has a 22 amino acid hydrophobic peptide that is known to act as a Afusion peptide@ during sperm-egg fusion. The mature fertilin beta subunit has an amino-terminal 90-93 amino acid domain that has sequence homology with a class of known integrin ligands (the snake venom disintegrins), and it is suggested that this domain plays a pivotal role in sperm-egg binding as an integrin-disintegrin interaction. Based on these observations, fertilin and the egg receptor for fertilin could be a potential target for contraception. Some scientists have explored the possibility of using purified fertilin as an antigen for immuno-contraception. Three male guinea pigs received injections of fertilin showing that 100% of the males were infertile. Several control males received injections of adjuvent without protein and none of them were inf erti le. Other studies showed that fertilin beta is expressed only in testis in humans. According to the previous work, we want to design a NH2 terminal 93 amino acid fragment of human fertilin beta (fertilin beta-NTP93) that utilizes the integrin-disintegrin binding domain as an antigen of vaccine and observe the inhibiting ability of sperm-egg binding by anti-human fertilin ?-NTP93 antibody in vitro. FUNDING Mellon Foundation-CONRAD Program (#MFG-97-34) PUBLICATIONS Moreno RD, Ramalho-Santos J, Sutovsky P, Chang E, Schatten G. Molecular membrane marker dynamics during rhesus spermatogenesis Acrosome biogenesis, Golgi apparatus and mitochondrial differentiation. Mol Biol Cell 9:66a, 1998. Ramalho-Santos J, Moreno RD, Sutovsky P, Wessel G, Schatten G. Membrane fusion and fertilization Implications for the presence of SNARE proteins on mammalian spermatozoa. Mol Biol Cell 9:332a, 1998.

Thesaurus Terms:
biomedical equipment development, inborn metabolism disorder, laser, microscopy, mother /infant health care, reproductive system, women's health animal tissue, bioimaging /biomedical imaging, clinical research, technology /technique

Institution: OREGON HEALTH & SCIENCE UNIVERSITY
PORTLAND, OR 972393098
Fiscal Year: 2000
Department:
Project Start: 01-MAY-1978
Project End: 30-APR-2004
ICD: NATIONAL CENTER FOR RESEARCH RESOURCES
IRG:

Abstract: The recent and unanticipated successes in both cloning sheep using nuclei from adult fibroblasts and in solving many forms of infertility by intracytoplasmic sperm injection (ICSI) now forces questions as to whether a combination of these approaches will result in reliable and efficient applications for propagating valuable research models, especially non-human primates. Spontaneous mutations resulting in unique animals to investigate diseases (e.g., retinitis pigmentosa) are periodically discovered in non-human primate colonies, and the ability to propagate these animals is vital for future investigations. While electrofusion of blastomere nuclei with enucleated rhesus oocytes resulted in two offspring (one male and one female) in 1996, last year=s attempts have not been successful. Intracytoplasmic nuclear injection (ICNI) holds bright promise as an alternative approach for propagating identical non-human primates. This project seeks to sequentially perfect IC NI c apabilities and develop new animal resources by addressing four specific aims. 1. Is intracytoplasmic sperm injection an effective and efficient method for propagating rhesus monkeys? 2. Is round spermatid injection (ROSI) an effective method for propagation? 3. Is intracytoplasmic nuclear injection, using blastomere nuclei obtained from rhesus monkey embryos injected into unfertilized enucleated oocytes, an effective and efficient method of propagation? 4. Is intracytoplasmic nuclear injection, using nuclei obtained from somatic cells cultured from adult and fetal rhesus tissues and injected into unfertilized enucleated oocytes, an effective and efficient method of propagation? ICNI may well find fantastic uses as a reliable, routine approach for propagating invaluable research animal resources of vital importance for the entire biomedical research community. FUNDING NIH R24 RR13632, HD18185 PUBLICATIONS Dominko T, Hewitson L, Simerly C, Luetjens M, Sutovsky P, Takahashi D, Schatten G. What is transferred during nuclear transfer? Imaging of mitochondrial, DNA, and centrosome dynamics in nuclear transfer rhesus embryos. Biol Reprod 58:99, 1998. Hewitson L, Takahashi D, Dominko T, Simerly CR, Schatten G. Assessing intracytoplasmic sperm injection Fertilization, embryo development, embryo transfer and pregnancy in the rhesus monkey. Fertil Steril 70:S242, 1998. Hewitson L, Takahashi D, Dominko T, Simerly C, Schatten G. Fertilization and embryo development to blastocysts after intracytoplasmic sperm injection in the rhesus monkey. Hum Reprod 13:3449-3455, 1998.

Thesaurus Terms:
genetic manipulation, inborn metabolism disorder, laser, microscopy, model design /development, reproductive system, transgenic animal, women's health animal tissue, bioimaging /biomedical imaging, clinical research, technology /technique

Institution: OREGON HEALTH & SCIENCE UNIVERSITY PORTLAND, OR 972393098
Fiscal Year: 2000
Department:
Project Start: 01-MAY-1978
Project End: 30-APR-2004
ICD: NATIONAL CENTER FOR RESEARCH RESOURCES
IRG:

Abstract: The overall goal of this grant is to increase our understanding of the etiology of Functional Hypothalamic Amenorrhea (FHA) and to test several novel treatment regimens for this reproductive disorder. Anovulation is the most common cause of infertility in women and FHA is the most common cause of anovulation. The proximate cause of FHA is insufficient hypothalamic gonadotropin-releasing hormone (GnRH) secretion. However, a constellation of neuroendocrine secretory disturbances attributable to altered hypothalamic function exist. Our current investigation in women and in monkeys implicates dysfunctional attitudes, psychosocial stress, and associated behaviors such as calorie restriction and exercise in the genesis of characteristic hypothalamic-pituitary-adrenal (HPA), HP-thyroidal (HPT), and HP-ovarian (HPO) secretory alterations. Thus, we conceptualize FHA as a state of altered hypothalamic homeostasis caused by synergism between psychogenic challenge and meta boli c compromise. Building on these findings, we hypothesize that cognitive behavior therapy (CBT) aimed at improving attitudes and correcting problematic behaviors will reverse hypothalamic derangements and restore ovulation. One aim of this grant is to test this hypothesis. However, because not all women with FHA will respond to CBT, we are continuing our quest to discern in women and in monkeys the neurobiological mechanisms underlying the synergism between psychogenic and metabolic stressors with the goal of identifying promising pharmacologic interventions. To refine our model of the pathogenesis of FHA, monkey studies are critical to identifying causality and pharmacologic options, while the human studies are integral to testing the efficacy of psychosocial and pharmacologic therapies. An interdisciplinary team of established investigators is working on these combined clinical and basic studies, with expertise in the areas of psychiatry, behavioral medicine, exercise physiology, neurobiology, and reproductive endocrinology. New findings this year indicate that monkeys with elevated basal heartrate are more prone to develop stress-induced reproductive dysfunction than monkeys with lower basal heartrate. We are currently examining whether this also holds true for women. FUNDING NIMH MH 50748-05 PUBLICATIONS Cameron JL, Bridges MW, Graham RE, Bench L, Berga SL, Matthews K. Basal heartrate predicts development of reproductive dysfunction in response to psychological stress. In The Endocrine Society Program and Abstracts 80th Annual Meeting (held in New Orleans, LA, June 24-27, 1998), p 138 (abstract #P1-76).

Thesaurus Terms:
endocrine gland /system, nervous system, psychology, reproductive system, women's health Mammalia, behavioral /social science research tag, clinical research

Institution: OREGON HEALTH & SCIENCE UNIVERSITY
PORTLAND, OR 972393098
Fiscal Year: 2002
Department:
Project Start:
Project End:
ICD: NATIONAL CENTER FOR RESEARCH RESOURCES
IRG:

Abstract: The overall goal of this project is to define the factors underlying the blunted growth hormone (GH) response to pharmacological stimulation that is characteristic of children and adults with depression and to utilize this information to identify neural systems which may underlie the development of affective disorders. Blunted stimulation of the anterior pituitary hormone, GH, after a number of pharmacological stimuli, occurs in patients with depression throughout the lifespan, and the blunted GH response is one of the most consistent physiological measures documented in children and adults with depression. Children with depression tend to exhibit less exploratory behavior and more inhibited behavior in a variety of psychological tests. We are performing the current study in the monkey to determine (1) if inhibited affect and blunted GH responsiveness are linked traits, and (2) if inhibited affect and blunted GH responsiveness predispose an individual to the development of anxious and /or depressive behaviors. Therefore, this project is quantifying the behavioral characteristics of young monkeys with regard to exploratory versus inhibitory behavior, and quantifying GH responsiveness to stimulation, and then assessing the degree of correlation between these parameters. Over the past 22 years, we developed 4 behavioral testing paradigms, based directly on paradigms used in children to assess these characteristics. In 1999, we tested 110 monkeys and we found a strong correlation between blunted GH responsiveness and A low reactivity@ in the playroom and stranger approach test s. Thus, we have evidence that at least in some individuals, these traits are linked. FUNDING NIMH MH41712 PUBLICATIONS Coleman K, Dahl RE, Ryan N, Cameron JL. Behavioral inhibition A new look at a familiar trait. Soc Neurosci Abstr 24:525, 1998 (abstract #211.1). Cameron JL, Coleman K, Bench LM, Sabatini M, Owenby T, Kupfer DJ. Differential development of anxious and depressive behaviors in rhesus monkeys dependent on the timing of maternal separation. Soc Neurosci Abstr 24:526, 1998 (abstract #211.7).

Thesaurus Terms:
endocrine gland /system, growth /development, mental disorder, nervous system, psychology Mammalia, behavioral /social science research tag

Institution: OREGON HEALTH & SCIENCE UNIVERSITY
PORTLAND, OR 972393098
Fiscal Year: 2002
Department:
Project Start:
Project End:
ICD: NATIONAL CENTER FOR RESEARCH RESOURCES
IRG:

Abstract: The overall goal of this grant is to experimentally test the hypothesis that the pubertal period represents a Awindow of heightened sensitivity@ to external perturbations that may have long-term health consequences. Specifically, this project is determining whether monkeys going through the pubertal transition are more sensitive to the various health consequences of social stress and moderately vigorous exercise training than monkeys in the early post-pubertal period. The health consequences that are being monitored are (1) reproductive function, including menstrual cyclicity and reproductive hormone secretion, (2) bone structure and growth, (3) body fat distribution, (4) serum lipid content, (5) glucose tolerance, (6) immune function, (7) susceptibility to viral infection, and (8) cardiovascular function, including resting heart rate, heart rate response to stress, and echocardiographic measurements. These parameters are being studied because they are known to be i nfluenced by social stress and exercise and they are all thought to be affected by reproductive steroid hormone levels. Thus, they are likely to be affected by the pubertal transition as well as by the external perturbations that are being studied. FUNDING The John D. and Catherine T. MacArthur Foundation PUBLICATIONS Rogers CJ, Brissette-Storkus CS, Hayes LA, Cameron JL, Chambers WH. Selective reduction in CD2 expression on CD2bright/CD8+ lymphocytes from cynomolgus monkeys (Macaca fascicularis) in response to acute stress. J Neuroimmunol 86:61-72, 1998. Rogers CJ., Brissette-Storkus CS, Chambers WH, Cameron JL. Acute stress-induced increase in NK cell granule content in monkeys. Soc Neurosci Abstr 24:1862,1998 (abstract 739.2). Rogers CJ, Brissette-Storkus CS, Cameron JL, Chambers WH. Acute stress alters both NK cell conjugation and lytic granule content in monkeys. Society for Natural Immunity Abstract, 1998.

Thesaurus Terms:
carbohydrate, cardiovascular system, communicable disease, diabetes mellitus, endocrine gland /system, growth /development, immunology, inflammation, nervous system, pancreas, physical fitness, psychology, reproductive system, skeletal system, women's health Mammalia, behavioral /social science research tag, clinical research

Institution: OREGON HEALTH & SCIENCE UNIVERSITY
PORTLAND, OR 972393098
Fiscal Year: 2002
Department:
Project Start:
Project End:
ICD: NATIONAL CENTER FOR RESEARCH RESOURCES
IRG:

Abstract:The overall goal of this study is to elucidate the physiological mechanisms by which short term changes in food intake lead to changes in the central neural drive to the reproductive axis in primate species. Previously, we have shown that brief periods of fasting (i.e., one to two days) lead to a significant suppression of luteinizing hormone (LH), follicle stimulating hormone (FSH) and testosterone secretion in adult male rhesus monkeys and men. Subsequent refeeding of monkeys after a brief period of fasting leads to a rapid and dramatic increase in LH and testosterone secretion within the first hour after meal intake. In the current grant, the first specific aim is to identify neuronal systems which may mediate the changes in LH secretion caused by fasting and refeeding. Experiments are examining the role of three specific neural systems in mediating nutrition-induced changes in LH secretion (A) the vagus nerves, (B) noradrenergic pathways, and (C) neuropeptid e Y (NPY)-containing neurons. The second specific aim is to determine the role of insulin and thyroid hormone (T3) in causing the suppression of LH secretion during fasting and the restoration of LH secretion during refeeding. The third specific aim is to determine the extent to which the diurnal pattern of LH secretion in the male rhesus monkey is determined by the pattern of daily food intake, and the timecourse with which changes in the pattern of food intake modify the pattern of LH secretion. Information provided by these studies is providing insight into the mechanisms by which reproductive function is suppressed in normal childhood, anorexia nervosa, bulimia nervosa, and with vigorous exercise training, in that these are all conditions where metabolic signals have been proposed to be at least partially responsible for the quiescent state of the reproductive axis. Studies completed this year have shown that NPY neurons throughout the hypothalamus are rapidly activated after miss ing a single meal, thus are in an ideal position to modulate changes in GnRH neuronal activity. FUNDING NICHD HD26888 PUBLICATIONS Caston-Balderrama AL, Cameron JL, Hoffman GE. Immunocytochemical localization of fos in perfused nonhuman primate brain tissue fixation and antisera selection. J Histochem Cytochem 46:547-556, 1998. Cameron JL. Fasting and reproduction in non-human primates. In Pennington Center Nutrition Series Nutrition and Reproduction (W Hansel, G Bray, DH Ryan, eds). Baton Rouge University of Louisiana Press, pp 95-109, 1998.

Thesaurus Terms:
endocrine gland /system, nervous system, nutrition, reproductive system, women's health Mammalia, clinical research, nutrition related tag

Institution: OREGON HEALTH & SCIENCE UNIVERSITY
PORTLAND, OR 972393098
Fiscal Year: 2002
Department:
Project Start:
Project End:
ICD: NATIONAL CENTER FOR RESEARCH RESOURCES
IRG:

Abstract: Depression and anxiety adversely affect millions of individuals, causing significant mortality and morbidity. Clinical studies over several decades suggest that these behavioral disorders emanate from both genetic factors and exposure to environmental factors. The interrelationships between genetic predisposition to these disorders, and how environmental and social factors modify genetic predisposition is critical to the development of clinical strategies for preventing these devastating disorders. For this purpose we are using a monkey model, to examine how social stress during early development triggers anxious and depressive behaviors and how social support modulates the development of these behaviors. Infant Japanese macaques are being tested early in life for temperament and growth hormone responsiveness. Clinical studies show a correlation between inhibited temperament and blunted GH responsiveness, and the anxiety and depressive disorders. Infant monkey s are then separated from their mothers and reared in a normal social colony environment. Behavioral studies are being used to determine if infants who receive greater social support have a decreased probability of developing anxious and depressive behaviors, compared to infants who receive less social support.FUNDING Center-supported project PUBLICATIONS None

Thesaurus Terms:
growth /development, mental disorder, nervous system, psychology, women's health Mammalia, behavioral /social science research tag, clinical research

Institution: OREGON HEALTH & SCIENCE UNIVERSITY
PORTLAND, OR 972393098
Fiscal Year: 2002
Department:
Project Start:
Project End:
ICD: NATIONAL CENTER FOR RESEARCH RESOURCES
IRG:

Abstract: The overall goal of this study is to determine to what extent common life stresses (i.e., dieting and psychological stress) impair fertility. It has been known for years that numerous forms of stress, including undernutrition, exercise, and psychological stress, can impair reproductive hormone secretion and can lead to decreased gonadal function, and impaired fertility, when the level of stress is severe. However, over the past several years, it has been shown for several different stresses (including both metabolic and psychological stresses) that mild, relatively acute forms of stress can also suppress reproductive hormone secretion; but we do not know if the effects of such stresses are great enough to actually impair fertility. These mild stresses are common in women's lives and if they play a role in impairing fertility, they could be a major factor contributing to infertility in our society. The projects in this research use a non- human primate model to determine if mild stress impact on fertility, and to study if the mechanisms underlying the suppression of reproductive hormone secretion my mild stress. A clinical component of this project begins to examine the role that mild stresses may play in the success rate of infertility treatment in women. Using this combined basic and clinical approach, the specific aims of this project are first, to define the impact of mild stress exposure on pregnancy rates in rhesus monkey; second, to identify the neural circuits which are responsible for mediating impairment of reproductive hormone secretion by mild stresses using a combined approach of pharmacological experiments and in vitro studies utilizing immunocytochemistry, in situ hybridization and confocal microscopy; and third, to determine if success rates in treating patients with tubal infertility by in vitro fertilization/embryo transfer (IVF-ET) are correlated with patient stress parameters. Together, the results of these studies will greatly increase our understanding of the role that common life stresses have on fertility in females, and will determine the underlying mechanisms whereby such stresses suppress reproductive function, providing information necessary for the future development of successful treatments for stress-induced infertility.

Thesaurus Terms:
hormone biosynthesis, reproduction, reproductive hormone, stress biological model, cooperative study, embryo /fetus transplantation, endorphin, environmental stressor, gamma aminobutyrate, in vitro fertilization, luteinizing hormone, menstrual cycle, neuroanatomy, neuropharmacology, progesterone, psychological stressor, steroid hormone receptor Macaca mulatta, behavioral /social science research tag, clinical research, confocal scanning microscopy, female, human subject, immunocytochemistry, in situ hybridization

Institution: OREGON HEALTH & SCIENCE UNIVERSITY
PORTLAND, OR 972393098
Fiscal Year: 2003
Department:
Project Start:
Project End:
ICD: NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
IRG:

OLINE RONNEKLEIV

Abstract: The objective of these experiments is to elucidate the effects of fetal cocaine exposure on neuronal development. Infants which are born to mothers who use cocaine, have increased risk of neurological and neurobehavioral deficiencies. These deficits imply that dopamine neurotransmission is altered. In adult animals cocaine inhibits reuptake of monoamines and thus, affects dopamine neurotransmission. However, very little is known about the effects of cocaine on developing neurons. In order to characterize cocaine-induced changes in brain development, we will focus on the dopaminergic neuronal systems, and explore the effects of cocaine on the molecular, cellular and structural changes that occur during the early stages of neuronal proliferation and migration. The first series of experiments will examine the effects of cocaine on the early ontogeny of tyrosine-hydroxylase (TH)-containing neurons destined to form the mesolimbic, nigrostriatal and tuberoinfundibular dopaminergic neuronal groups. We will determine the time of origin, anatomical location and maturation of cells containing TH within the ventral tegmental/substantia nigra and hypothalamic areas, and the development of their fiber projections using immunocytochemistry and in situ hybridization. Secondly, we will explore the effects of cocaine on the ontogeny of dopamine receptors in the nigrostriatal/mesolimbic and hypothalamic systems. We will detect and quantify D-1 and D-2 receptor mRNAs and measure receptor binding in the nucleus accumbens/striatum and the hypothalamus during the early phase of dopaminergic neuronal development. These experiments will be performed using in situ hybridization and polymerase chain reaction (PCR) to localize and quantify and mRNAs, and receptor autoradiography to quantify receptor binding. The effects of cocaine on the development of the dopamine transporter will also be investigated using [3H]CFT (WIN 35,428) as a ligand. The next series of experiments will examine the effects of cocaine on the ontogeny of dopaminergic target neurons, focusing on enkephalin neurons in the nucleus accumbens and the striatum, and on GnRH neurons in the hypothalamus. These experiments will be performed using immunocytochemistry and in situ hybridization to study the maturation of peptide expression and migration. Lastly, we will quantify proenkephalin mRNA and GnRH mRNA during peak migration and during the post migration period in control and cocaine- treated fetuses, using PCR. The results from these studies will provide important information about normal development and the effects of cocaine on major dopaminergic neuronal systems and their target neurons. Therefore, information will be generated to understand the basic effects of cocaine on developing neurons. This data can be used to develop treatment strategies to counter the effects of cocaine in infants and young children.

Thesaurus Terms:
cocaine, developmental neurobiology, dopamine receptor, drug abuse, embryo /fetus toxicology, neurogenesis G protein, cAMP response element binding protein, dopamine transporter, dynorphin, enkephalin, gamma aminobutyrate, longitudinal animal study, maternal behavior, neural transmission, opioid receptor, prosencephalon, transcription factor, tyrosine 3 monooxygenase Macaca mulatta, autoradiography, immunocytochemistry, in situ hybridization

Institution: OREGON HEALTH & SCIENCE UNIVERSITY
PORTLAND, OR 972393098
Fiscal Year: 2000
Department: NONE
Project Start: 15-DEC-1991
Project End: 31-JAN-2002
ICD: NATIONAL INSTITUTE ON DRUG ABUSE
IRG: NIDA

ELIOT SPINDEL

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, 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 972393098
Fiscal Year: 2000
Department: NONE
Project Start: 01-FEB-1999
Project End: 31-JAN-2004
ICD: NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
IRG: LBPA

Abstract: The goal of this project was to obtain preliminary evidence for the development of a nonhuman primate model of polycystic ovarian syndrome (PCOS). The 4 monkeys used for the experiment had cycled consistently over the previous 12 months (12 or 13 menses recorded, at regular intervals). They were provided with an intraovarian graft of genetically modified baby hamster kidney cells, encapsulated in a polymer of poly [acrylonitrile vinyl chloride, P(AN-VC)]. The surgical implants were performed as close to the first day of the new menses cycle as possible. Two of the monkeys received devices which contained NGF-secreting cells and two of the monkeys received control devices containing unmodified cells. The implants were 0.7 mm outside diameter and 7 mm long. Each monkey received two implants in each ovary. Following laparotomy to expose the ovaries, the implants were inserted into the ovary using a large bore needle and plunger (a suture in the ovarian capsule prevented migration of the device). Following implantation, all four monkeys continued to cycle and did so throughout the six month study; thus, the implants themselves did not affect normal ovarian function. The collected serum samples were assayed for ovarian steroids and luteinizing hormone. Of the steroids analyzed (estradiol, progesterone, testosterone and androstenedione), only androstenedione was elevated following the grafting of NGF producing cells; the other steroids remained constant. Luteinizing hormone levels were also elevated in the monkeys that received the NGF producing grafts. Androstenedione and LH are the two hormones most consistently linked with PCOS in humans. The ovarian morphology following grafting is still being assessed; however, initial observations suggest a possible increase in the number of follicular structures, again reminiscent of PCOS. Although further analysis of the data is necessary, preliminary evaluation suggests that an elevation of intraovarian N GF levels may be one of the components required to generate a nonhuman primate model of PCOS. FUNDING Center-supported project PUBLICATIONS None

Thesaurus Terms:
endocrine gland /system, gene therapy, growth /development, nervous system, reproductive system, women's health Mammalia, animal tissue, clinical research

Institution: OREGON HEALTH & SCIENCE UNIVERSITY
PORTLAND, OR 972393098
Fiscal Year: 2000
Department:
Project Start: 01-MAY-1978
Project End: 30-APR-2004
ICD: NATIONAL CENTER FOR RESEARCH RESOURCES
IRG:

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, 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 972393098
Fiscal Year: 2002
Department: NONE
Project Start: 01-JUL-2002
Project End: 30-JUN-2007
ICD: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
IRG: HED

Abstract: Reproductive senescence in the female rhesus (18-22 years old) was monitored by analysis of serum estrogen (E), progesterone (P) and LH. While many animals had normal menstrual cycles, a subpopulation displayed irregular cycles or were acyclic with low E and P with elevated LH. This corroborates suggestions that menopause occurs in the mid-twenties. Although the macaque hippocampus is believed to be regulated by ovarian steroids, previous studies failed to detect estrogen receptors (ER-?). To see if the ? form of ER is present in this region, macaque cDNAs for ER-? were isolated and used for ISH. High levels of ER-? were indeed found throughout the monkey hippocampus. Ovarian steroid effects were also detected in the prefrontal cortex. ICC analyses showed an increase of dopamine beta hydroxylase (DBH) and serotonin upon ovariectomy. Immunoreactivity for choline acetyl transferase was unchanged. Replacement with E reversed the effects of ovx on DBH whereas E +P decrea sed serotonin fiber density. In vivo brain imaging was performed using Magnetic Resonance Imaging (MRI) to examine structural changes in aged male monkeys. Image analysis confirmed an age-related increase in the total lateral ventricular volume in a subpopulation of aged animals. The increase in ventricular volume may be predictive of age-related behavioral deficits. The initial phase of participation in the NIA Biomarkers of Aging database has shown that longitudinal changes in animals from this Center, in general, agree with compiled data from other participating Centers. Expansion of this effort is currently being discussed. The establishment of reliable Biomarkers of Aging, such as some blood chemistry values, will be valuable for establishing normative ranges for aged monkeys. This database will also be useful for evaluating the acute effects of experimental manipulations thought to influence the rate of aging and lifespan. FUNDING Nonfederal institutional funds PUBLICATIONS Kohama, SG, Rodrigues SM, Pau C, Urbanski HF. Age-related change of glutamate receptors and neuropathology in the monkey prefrontal cortex. Soc Neurosci Abstr 24:464, 1998 (abstract 179.22).

Thesaurus Terms:
aging, nervous system Mammalia

Institution: OREGON HEALTH & SCIENCE UNIVERSITY
PORTLAND, OR 972393098
Fiscal Year: 2000
Department:
Project Start: 01-MAY-1978
Project End: 30-APR-2004
ICD: NATIONAL CENTER FOR RESEARCH RESOURCES
IRG: