Mark E. Wilson, Ph.D. - US grants

Puberty in mammals is controlled by a complex interplay of endocrine and environmental variables. We propose a longitudinal series of studies to examine the hormonal parameters of sexual maturation in female rhesus monkeys, and to determine how social and physical characteristics of the environment regulate this process. Intact and ovariectomized females treated with estradiol (E2) will be studied from 12 months of age (prepuberty) through 48 months of age (young adult) with specific experiments demarcated at critical times during development. We will determine if maturation in this primate is characterized by a gradual decrease in the ability of E2 to inhibit the secretion of luteinizing hormone (LH) and follicle-stimulating hormone prior to first ovulation. Developmental changes inthe pulsatile secretion pattern of LH will be assessed to determine their precise temporal relationship to (1) the predicted decrease in E2 negative feedback on gonadotropin secretion, (2) menarche, and (3) the capacity to ovulate. As rhesus monkeys are seasonal breeders, attempts to induce on ovulatory surge of LH by E2 treatment at specific times during development will determine how age and annual photoperiodic changes control the occurrence of first ovulation. Furthermore, an examination of changes in serum patterns of adrenal androgens and growth hormone will determine how these hormones synergistically influence the decrease in E2 negative feedback on gonadotropin secretion and pubertal growth rates. Demographic variables also will be assessed to determine their influence on the hormonal progression of puberty and the outcome of pregnancy associated with first ovulation. The proposed experiments will elucidate the hormonal sequence of puberty in a primate species, and will thus provide a better understanding of how the brain and ovary interact to produce a reproductively viable adult. The examination will further clarify normal and abnormal pubertal development in humans.

The polymorphonuclear neutrophil plays a prominent role in host resistance to foreign bacteria, including organisms involved in the development of periodontal disease. This is evidenced by the fact that patients with a number of different intrinsic neutrophil defects often present with particularly severe forms of periodontitis. In addition, pathogenic bacteria, including those from the oral cavity, can produce products which may either enhance or depress host immune responses. One such example of the latter case is the recently described low molecular weight substance present in Capnocytophaga culture supernatants which inhibits neutrophil chemotaxis. An important aspect of the proposed study will be to isolate this chemotaxis inhibitor and to characterize its chemical properties. We have also proposed studies to examine whether this substance, or other substances present in Capnocytophaga culture supernatants, influences other aspects of neutrophil function. Finally, we will characterize the interaction between selected Capnocytophaga strains and host mediation (antibody and/or complement) systems which facilitate clearance of these bacteria, either directly or cooperatively with neutrophils. These studies will provide fundamental information concerning a) mechanisms whereby oral bacteria may be effectively neutralized by host mediation systems and/or phagocytic cells and b) modulation of host responses by bacterial products which may contribute to the persistence of oral pathogens.

The period of postpartum infertility in primiparous female primates is longer than the interval in multiparous females. First pregnancy and resultant lactation typically occur in female primates prior to the completion of the growth phase and therefore have implications for continued growth and subsequent reproductive capacity. Pregnancy and lactation produce specific changes in the neuroendocrine system controlling ovulation, and also increase the nutritional requirements of the female, both effects which may be exacerbated in young, sexually-mature females. We propose a series of longitudinal experiments using four groups of female rhesus monkeys to determine how age, parity and maternal nutritional status influence the duration and characteristics of postpartum infertility. Subjects will be studied in provisioned, outdoor housed social groups to assess how mother-infant interactions influence the lactational process. We will determine by measurement of body weight, percent body fat, and skeletal growth how age at first pregnancy and lactation influence the completion of the growth process. The studies will specify how first pregnancy and first lactation differentially influence the pattern of growth. A comparison of multiparous females to different-aged primaparous females during the lactational period will determine how age and parity affect patterns of 1) maternal care, 2) infant nursing behavior and physical growth, and 3) maternal endocrine status, measured by serum levels of prolactin, luteinizing hormone, follicle-stimulating hormone, and ovarian steroids. These analyses will elucidate how the duration of postpartum infertility in primiparous mothers is influenced by the nutritional costs of lactation, by infant nursing patterns, or by the interaction between these two variables. Finally, we will determine how parity and age influence the resumption of fertility and the capacity of females to conceive and maintain pregnancy. The characteristics of first lactation, including alterations in maternal nutritional status and basal insulin levels, will be related to the subsequent reproductive ability of the female. This project will further define how pregnancy and lactation influence the completion of growth and the parameters of subsequent reproductive activity in young primate mothers.

The duration of lactational infertility is significantly lengthened in young primate mothers. We hypothesize that altered neuroendocrine regulation of ovulation in adolescents synergizes with the nursing-induced inhibition of ovarian function to greatly reduce the reproductive capacity in young females. A series of longitudinal studies will evaluate the hypotheses that (1) nursing suppresses ovarian function by suckling-induced decreases in gonadotropin-releasing hormone (GnRH) secretion, and thus luteinizing hormone (LH) and follicle stimulating hormone (FSH) and (2) sexually mature adolescent females are more responsive to estradiol (E2) negative feedback and nongonadal restraint of LH and FSH release. The use of seasonal breeding rhesus monkeys will identify the biobehavioral factors which maintain lactational infertility at a time which is optimal-fbr the occurrence of ovulations in nonlactating females. We will determine how quantifiable changes in nursing behavior alter the neuroendocrine control of ovarian function. The resilience of episodic LH and FSH release and pituitary sensitivity to GnRH will be assessed in adult and adolescent mothers following removal of the suckling stimulus during mid and late lactation. Pulsatile infusion of one of three doses of GnRH to reinitiate ovulatory cycles will evaluate the hypothesis that lactation disrupts endogenous GnRH secretion and will determine if adolescent mothers are less responsive to this treatment. These studies will also assess the direct effects of nursing on ovarian function in the presence of appropriate LH and FSH support. Further dose-response studies will determine if suckling and adolescent status affect LH and FSH secretion by enhanced E2 negative feedback or the induction of nongonadal restraint. An examination of periparturitional levels of prolactin (PRL) will determine if these are predictive of eventual neuroendocrine deficits in adolescent mothers. Also, studies will elucidate how lactation and adolescence diminish ovulatory capacity during the breeding season by assessing the effects of low intensity nursing on episodic LH and FSH release and subsequent ovarian responsiveness. Finally, the behavioral and physiological antecedents which initiate changes in nursing behavior will be described. The manipulation of both nursing behavior and serum PRL levels will determine the differential contribution of each variable to the maintenance of lactation infertility. These studies will describe the basic mechanisms which regulate lac- tational infertility in primates and how it is prolonged in adolescent mothers.

Dr. Wallen has developed a model that integrates the central effects of hormones with peripheral mechanisms affecting their metabolic fate that has direct relevance to the development of human steroid replacement therapies. From work with humans and primates, it is clear that removal of female ovaries or damaging their ability to function drastically reduces sexual interest and satisfaction. A critical problem is which steroid hormone or combination of hormones can mimic the action of the ovaries and adrenals on these behavior. This is further complicated since blood of female rhesus and human females contains special proteins that can tightly bind estrogens and androgens. When hormones are attached to these proteins, called sex hormone binding globulins or SHBG, they cannot affect the nervous system and are inactive. When they become unattached from the protein they are once again active. Although both steroid hormones bind to SHBG, androgens bind more tightly than estrogens. Dr. Wallen is examining whether the binding of androgens to these special blood proteins frees estrogens to influence behavior. His work will clearly define the role of both estrogens and androgens. Furthermore, finding that SHBG strongly influences the behavioral effectiveness of ovarian and adrenal steroids will markedly change current views of hormone action. The outcome of his innovative work will provide new information on how steroid hormones interact to modulate their individual dynamics and effectiveness and could lead to the development of better hormonal treatments for humans with endocrine disorders.

The signals responsible for the development of gonadotro

Risk for development of localized juvenile periodontitis (LJP) varies markedly among different e c groups, with blacks being roughly 15 times more likely than Caucasians to develop this disease. LJP also tends to aggregate within families, suggesting that susceptibility may be genetically-defined. To date, however, a genetic marker for LJP has not been identified. Black LJP subjects mount a strong IgG response toward A. actinomycetemcornitans, the etiologic agent of this disease, a significant proportion of the IgG response being comprised of antibodies of the IgG2 subclass. The effectiveness of these IgG2 antibodies in host defense has not been defined. Genetically- defined polymorphism of IgG Fc receptors (Fc-lambda-R) can markedly affect receptor affinity for human IgG subclasses, particularly IgG2. A principal objective of this proposal is to evaluate the relationship between Fc-lambda-R polymorphism, opsonic activities of IgG subclass antibodies to A. actinomycetemcomitans, and ethnic variations in risk for LJP. Moreover, we will examine the initial IgG subclass responses to this organism following the onset of disease. Specifically, we propose to: l. Determine if variations in the frequency of allelic polymorphism of Fc-lambda-R exist among UP subjects of diverse ethnic background. 2. Define the influence of Fc-lambda-R polymorphism upon the effectiveness of IgG subclass antibodies in LJP sera in promoting opsonization of A. actinomycetemcomitans. 3. Assess temporal changes in the IgG subclass responses to A. actinomycetemcomitans which occur during the early stages of transition from a periodontally healthy state to one of LJP. 4. Compare the IgG subclass responses of ethnically diverse LJP subjects to key antigens of A. actinomycetemcomitans. The information derived from the proposed studies will provide insight into the role of Fc-lambda-R polymorphism in defining risk for LJP among various ethnic groups, and in regulating the properties of IgG subclass antibodies to A. actinomycetemcomitans in sera of LJP subjects. Moreover, these studies will enhance our understanding of the dynamics of the IgG response to A. actinomycetemcomitans following the onset of LJP, as well as of qualitative and/or quantitative variations in the IgG response of ethnically diverse LJP subjects to this organism.

The neutrophil plays a key role in host defense against infection due to extracellular bacteria. Considerable evidence also indicates that the neutrophil is a critical element of host defense against periodontal injury initiated by bacteria in dental plaque. In order to provide such defense, adequate numbers of neutrophils must first penetrate vascular endothelium through an adhesion-dependent process. Following extra- vasation, neutrophils must migrate toward, ingest and ultimately destroy the target bacteria. Defects in neutrophil production or function often result in increased susceptibility to recurrent bacterial infection, and appear to predispose to early-onset, severe forms of periodontal disease which may arise prior to or during puberty. Susceptibility to severe periodontitis varies widely in the adult population. Cross-sectional studies conducted at the Clinical Research Center have identified certain variables associated with increased risk of severe periodontitis. Included in this group of "risk indicators" are diabetes and smoking. A key objective of studies proposed in this Clinical Research Center application is to conduct longitudinal studies necessary to establish if diabetes and/or smoking constitute true "risk factors" for development of severe periodontitis. Diabetes has long been associated with increased susceptibility to infection, attributable in part to altered neutrophil function. Neutrophil adherence, chemotaxis, phagocytosis and killing are often decreased in diabetes. Vascular changes observed in diabetes may also impede neutrophil extravasation into infected tissues. Various components of tobacco smoke have also been reported to inhibit neutrophil function. The objective of this pilot study is to determine whether apparent increases in susceptibility to periodontitis seen in diabetics or smokers are associated with impaired neutrophil function. Specific parameters to be evaluated will include a) in situ crevicular leukocyte responses to casein, b) expression of cell adhesion molecules (Mac-1/CR3, L- selectin) using anti-adhesion monoclonal antibodies in conjunction with flow cytometric techniques, c) in vitro aerobic and anaerobic bactericidal activity toward Actinobacillus actinomycetemcomitans, d) neutrophil degranulation, as determined by flow cytometric analysis of right angle light scatter, and e) chemotaxin (IL-8, fMLP)-induced signal transduction (IP3 production). Neutrophil responses will be compared in a) diabetic vs. non-diabetic periodontitis patients (balanced for smoking, microflora and age), b) smoker vs. non-smoker periodontitis patients (non-diabetic only balanced for microflora and age), and c) periodontitis subjects vs. periodontally healthy (diabetic and non- diabetic, as well as smoker and non-smoker) controls. These pilot studies are designed to provide the basis for an in-depth assessment of mechanisms by which smoking and/or diabetes impairs neutrophil-mediated defenses.

Growth hormone (GH) and its primary effector, insulin-like growth factor- I (IGF-I), govern the tempo and degree of adolescent growth. In addition to anabolic effects, the OH axis may regulate the hypothalamic -pituitary - ovarian unit (HPO) during puberty and affect ovarian physiology in adults. It is not clear whether the GH axis is obligatory for the normal progression of puberty and adult fertility and, if so, whether these effects are mediated through changes in H-P regulation of luteinizing hormone releasing hormone (LHRH) and LH and/or ovarian sensitivity. The proposed studies, using a female rhesus monkey model, will not only clarify the neuroendocrine regulation of the GH axis in adolescents and adults but will also determine whether the effects of GH and IGF-I on reproduction are limited to the puberty and how these may alter the HPO function. Age-dependent changes in pulsatile GH secretion will be assessed in intact and ovariectomized (OVX) adult and adolescent females treated with varying doses of estradiol (E2) to determine if the regulation of pulsatile OH by E2 is differentially affected by age. Additional studies will test the hypothesis that IGF-I differentially regulates pulsatile GH in adult and adolescent females and under specific E2 treatment conditions. Finally, the response to GH-releasing hormone (GHRH) will be assessed to determine if the age-dependent decline in serum OH results from decreased GHRH release and if this is modified by IGF-I and E2. In situ hybridization will quantify gene expression of H - P IGF-I and hypothalamic GHRH and somatostatin (SRIF) and autoradiography will quantify H - P receptor populations of these peptides to test the hypothesis that the decline in OH secretion with aging results from decreased GHRH and increased SRIF. Additional studies of these animals will examine the specific reproductive effects of the GH axis. The effects of IGF-I, a SRIF-analog (SRIFa), and IGF-I & SRIFa together on gonadal hormone secretion during spontaneous ovulatory cycles and following LHRH agonist treatment will test the hypothesis that IGF-I is critical for normal response to LH in adults. The addition, ovarian IGF-I and II gene expression and content of IGF-I and IGF binding protein-3 in follicular fluid will be quantified in adult and adolescent females treated with saline, IGF-I, SRIFa, or IGF-I & SRIFa to determine how these are regulated by age and alterations in peripheral IGF-I levels. Pulsatile LH secretion will be assessed in these adolescent and adult, intact and E2-treated OVX females to determine if the alteration in sensitivity to E2 negative feedback on LH is restricted to the peri- pubertal period and if IGF-I mediates such effects in adults. Administration of N-methyl-D, L- aspartic acid at specific reproductive stages and under specific E2 treatment conditions will assess how IGF-I affects LHRH neuronal activity. Finally, we will test the hypothesis that IGF-I prevents the decline in pulsatile LH observed following an acute fast thus affecting LHRH neuronal activity during catabolic states. These data will further our understanding of the physiological significance of the interactive effects of the GH axis and reproduction throughout the female lifespan.

The Oral Immunology/Microbiology Research Group was founded in 1991 as a means of promoting collegial interaction and collaboration among individuals interested in the immunology and microbiology of the oral cavity, particularly as related to oral disease (dental caries and periodontal disease). Formed in response to a general perception that large meetings such as the IADR/AADR General Sessions do not provide a forum suitable for intimate discourse and collaboration, the OIMRG is comprised of more than seventy investigators representing thirty-nine universities, research centers, and commercial organizations in the U.S. and abroad. Once each year the group convenes for an annual meeting which consists of three scientific session, each focusing upon a distinct area of oral immunology and microbiology. It is primarily, but not exclusively, through the annual meeting that the objectives of the Oral Immunology/Microbiology Research Group are achieved. These objectives include the following: 1. To foster interaction and collaboration among scientists interested in oral immunology and microbiology. 2. To promote information exchange and collaboration between academicians and their colleagues in the private sector who are engaged in basic and clinical studies pertaining to oral health and disease. 3. To provide a forum through which new investigator entering the fields of oral immunology/microbiology can begin to "network" with more established investigators and to establish contact with representatives of federal and non-federal agencies which may be a potential source of funding for future studies. The 7th annual meeting of the Oral Immunology/Microbiology Research Group has been scheduled for Feb. 21-24, 1997 and will be held at the Sheraton Charleston, in Charleston, SC. Session topics will include a) host responses to oral bacteria, b) mechanisms of microbial pathogenesis employed by oral bacteria, and c) mechanisms of host-mediated injury to oral tissues. Guest lecturers will discuss the role of IL-12 in infectious disease, as well as the use of in vivo expression technology as a means of studying bacterial virulence factors. Perpetuation of the OIMRG and its annual meetings greatly enhances interaction/collaboration among investigators with expertise in immunology and microbial pathogenesis as related to oral health and disease.

model design /development; growth /development; hormones; endocrine gland /system; Mammalia;

drug receptors; receptor binding; ligands; polymers; peptide chemical synthesis; protein sequence; peptides; molecular site; chemical binding; intermolecular interaction;

Estradiol (E2) is essential for the health, estradiol (E2) also normalizes behavioral systems related to mood and sexuality. Although E2 replacement prevents bone loss and reduces the risk of cardiovascular disease in hypo- estrogenic women, such treatments may adversely affect the breast uterus, despite co-treatment with progestins. Consequently, non-steroidal estrogens have been developed with specific targeted effects as E2 agonists in some tissues and antagonists in others. Since these tissue- specific actions are likely mediated through subtypes of the estrogen receptor (ER), the compounds are described as selective estrogen receptor modulators (SERMs). Despite their well-documented efficacy in peripheral target tissues, it is not known whether these SERMs act as E2 agonists or antagonists within the CNS regulating neuro-chemical mechanisms subserving complex social behavior. This project will determine if the SERMs, tamoxifen and raloxifene, are E2 agonists or antagonists in the neuroendocrine regulation of behavior in female rhesus monkeys. Using ovariectomized females, each Specific Aims will test the hypothesis that SERMs, in the absence of E2, act as agonists whereas, in the presence of E2, are antagonists. Preliminary studies will determine the pharmacokinetics of the SERMs to set the parameters of the treatment protocol. Specific Aim 1 will determine if SERMs mimic E2 and increase affiliative behavior mediated by increased central serotonergic (5HT) activity, assessed from concentrations of 5HT and its metabolite in cerebrospinal fluid, quantitative receptor autoradiography, and in situ hybridization for the 5HT re- uptake transporter. Subsequent studies will test the hypothesis that these SERMs antagonize endogenous E2, diminishing 5HT activity and increasing aggressiveness. Specific Aim 2 will determine if SERMs, in the absence of E2, induce female sexual motivation and coordinate gonadotropin synthesis and secretion. Additional studies will test the hypothesis that these SERMs antagonize endogenous E2, blocking the induction of sexual motivation and regulation of gonadotropin synthesis and release. These data will provide not only a better understanding of the biology of SERMs but will also help clinicians and patients evaluate of these compounds on behavioral health.

DESCRIPTION (Adapted from applicant's description): Acquisition of a critical body mass is thought to be important for the initiation of puberty. Although a number of studies in a variety of species have linked body size to puberty, the signal responsible for increasing pulsatile gonadotropin hormone releasing hormone (GnRH) secretion and initiating puberty is unknown. However, the recent identification of the protein leptin has lead to the hypothesis that a developmental increase in leptin, derived from accumulating fat mass, acts centrally to increase GnRI-1 secretion and a parallel increase in the pituitary release of the gonadotropins, resulting in gonadal activation. While the hypothesis that leptin initiates puberty has yet to be adequately tested in primates which, unlike rodents, show a protracted period of post-natal development prior to puberty. Before prospective studies can be initiated in primates, additional preliminary data derived from a female monkey model are needed to A) define precisely the temporal relationship between prepubertal increases in diurnal leptin concentrations and the emergence of nocturnal gonadotropin release and B) develop a protocol to assess how the sustained administration of leptin affects GnRH secretion in well nourished females. Specific Aim 1 will test the hypothesis that the developmental rise in nocturnal leptin secretion precedes the emergence nocturnal gonadotropin secretion in females. Specific Aim 2 will test the hypothesis that the timing of daytime food intake influences the pattern but not amplitude of the diurnal leptin secretion. Specific Aim 3 will determine the dose of sustained administration leptin that reverses the fasting-induced suppression of gonadotropin secretion and will test the hypothesis that leptin binding protein facilitates leptin's access to the brain and effects on neuroendocrine systems. Specific Aim 4 will obtain information on the efficacy of leptin administration to immature females on metabolic and reproductive parameters.

Although studies from a variety of species indicate that a structural or functional shift in neurochemical input into gonadotropin releasing hormone (GnRH) neurons induces the developmental increase in GnRH secretion initiating puberty, the factor(s) responsible for this change are not known. Using female rhesus monkeys, this project will test the hypothesis that a signal emanating from growth, namely insulin like growth factor (IGF)-I, changes the nature of this input, increasing GnRH and pituitary gonadotropin secretion, initiating puberty. The working hypothesis is that a prepubertal increase in IGF-I stimulates the emergence of GnRH secretion as assessed from nocturnal pulsatile gonadotropin release. Specific Aim 1 will test this hypothesis by using two different models of growth hormone (GH) insensitivity. A GH-IGF-I deficient model will be produced by treating juveniles with a GH receptor antagonist. It is predicted that the emergence of nocturnal gonadotropin secretion will be disrupted in GH antagonist-treated females compared with controls but replacement therapy with IGF-I will normalize this pattern. In the second study, reproductive maturation will be arrested by treating juveniles with an GnRH analog to prolong the prepubertal period of hypogonadotropism. Upon the cessation of analog treatment at an age equivalent to mid puberty in control females, it is predicted that co-treatment with a GH receptor antagonist will suppress the expression of the developing GnRH pulse generator, inferred from robust nocturnal pulses of gonadotropins, compared to females only treated with the analog. In contrast, this suppression of the GnRH pulse generator by GH antagonism will be reversed by co-administration of IGF-I. Specific Aim 2 will test the hypothesis that the integrity of the GH-IGF-I axis during the neonatal period is essential for the subsequent GH-IGF-I activity, prepubertal growth and the timing of puberty. Females treated neonatally (from birth-8 mo.) with a GH receptor antagonist will be compared to controls and to females treated neonatally with a GnRH analog, a treatment known to delay puberty in monkeys and rats. It is predicted that a postnatal disruption of GH-IGF-I will produce long term deficits in the GH axis, diminishing growth, delaying the emergence gonadotropin secretion and the onset on puberty. The data derived from these studies will provide significant new information on how the GH axis regulates puberty and, thus, a better understanding of aberrations in growth and development in children.

ethology; gonadotropin releasing factor; hormone regulation /control mechanism; Primates; animal colony;

structural biology; protein structure; biomedical resource;

endocrinology; Primates; animal colony;

neoplasm /cancer

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Core acquired a mass spectrometer with gas chromatography and HPLC capabilities. The Core has added this service to provide small molecule and proteomics that is not available through traditional immunoassay technologies. The Biomarkers Core Lab continues to provide steroid and protein hormone assay services for investigators. In 2009, the lab served 66 different investigators located throughout the country, including Emory (41%), other academic institutions (50%), government labs (3%), and private industry (6%). During the last year, the Lab processed 31,242 samples in support of research. Costs incurred in the performance of the assays are recovered using a charge back to all users that, in the past year, resulted in $322,707 recovered, a change of +30 and +40% above the amounts recovered for 2007 and 2008, respectively.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This project is designed to develop a protocols using liquid chromatography-mass spectroscopy to quantify changes in brain chemistry resulting from cocaine abuse using a monkey model and experimentally induced menopause using a mouse model. During the current year, levels of glutamate and GABA have been measured in microdialysis samples collected from the striatum of monkeys self-administering cocaine. In the second project, ovarian failure, to mimic the loss follicles characteristic of menopause, was induced in control mice and mice over-expressing the amyloid protein. Analyses will proceed in the coming year in the development of neuropathology in control versus experimental animals.

Animals; Aquadiol; Area; Binding; Binding (Molecular Function); Brain; Brain region; CRISP; Computer Retrieval of Information on Scientific Projects Database; Dimenformon; Diogyn; Diogynets; Encephalon; Encephalons; Estra-1,3,5(10)-triene-3,17-diol (17beta)-; Estrace; Estradiol; Estradiol-17 beta; Estradiol-17beta; Estraldine; Estrogen Receptors; Estrogenic Agents; Estrogenic Compounds; Estrogens; FES; FPS; FPS-FES Oncogene; Female; Fujinami Sarcoma Virus and Feline Sarcoma Virus Transforming Gene; Funding; Gene Action Regulation; Gene Expression Regulation; Gene Regulation; Gene Regulation Process; Grant; Hypophysis; Hypophysis Cerebri; Hypothalamic structure; Hypothalamus; Institution; Investigators; Lead; Ligand Binding; Ligands; Medical Imaging, Positron Emission Tomography; Molecular Interaction; Monkeys; NIH; National Institutes of Health; National Institutes of Health (U.S.); Nervous System, Brain; Nervous System, Pituitary; Oncogene FES; Oncogene, FPS-FES; Ovocyclin; Ovocylin; PET; PET Scan; PET imaging; PETSCAN; PETT; Pb element; Pituitary; Pituitary Gland; Positron Emission Tomography Scan; Positron-Emission Tomography; Progynon; Proton Magnetic Resonance Spectroscopic Imaging; Rad.-PET; Radiation Chemistry; Radiochemistry; Radiology; Radiology Specialty; Radiology, General; Research; Research Personnel; Research Resources; Researchers; Resources; Source; Therapeutic Estradiol; Therapeutic Estrogen; United States National Institutes of Health; density; design; designing; heavy metal Pb; heavy metal lead; hypothalamic; neuroimaging; neuropathology; new technology; radioligand; receptor binding

The Reproductive Status Core will be responsible for the collection of biological samples from women, chimpanzees, and rhesus monkeys to confirm classification of reproductive age, characterize gonadal steroid patterns within cycles, and assess menopausal status. Using previously validated immunoassays, reproductive hormone assessments will be made by Core personnel using the facilities provided by the Yerkes NPRC Endocrine Core Laboratory. Data will be transmitted to the Administrative Core and Project Leaders to schedule the cognitive testing in Projects 1 and 2 and neuroimaging in Project 3 for the early to mid follicular phase for pre- menopausal females and to quantify the gonadal hormone milieu at the time of the assessment for pre- and post menopausal females. An ovulatory cycle will be confirmed in young and middle aged females by a sustained, 7 to 10 day rise in progestin (serum progesterone for monkeys;urinary pregnanediol for chimpanzees;salivary progesterone for women). Short luteal phase ovulatory cycles will be identified by cycles with an abbreviated, 3 to 7 day, increase in progestins. The period of the sustained increase in progestins followed by menstruation will define the luteal phase. Anovulatory cycles will be identified by intermittent increases in estrogen (serum estradiol for monkeys;urinary conjugated estrogens for chimpanzees;salivary estradiol for women) in the face of persistently low and unvarying concentrations of progestins during the inter-menstrual period. The follicular phase of an ovulatory cycle will be identified as the period following menstruation characterized by rising concentrations of estrogen prior to the increase in progestin at ovulation. Menopausal status will be confirmed in aged rhesus monkeys by elevated serum levels of FSH in the face of reduced estradiol and inhibin B concentrations;persistently low and unvarying serum progesterone;and amenorrhea in rhesus monkeys;in aged chimpanzees by elevated urinary of FSH in the face of reduced conjugated estrogen;persistently low pregnanediol concentrations; amenorrhea;and reduced perineal swellings;and in older women by reduced salivary concentrations of estradiol and progesterone in the face of amenorrhea and self-reports of menopausal symptoms.

DESCRIPTION (provided by applicant): Postnatal stressors, producing a dysregulation of the limbic-hypothalamic pituitary adrenal (LHPA) axis, can have a deleterious effect on a child's pubertal and psychosocial development. Importantly, the gonadal release of estradiol (E2) occurring during the pubertal transition likely contributes to the continued maturation of cortico-limbic circuits that regulate emotional behavior but stress-induced delayed puberty could compromise this development. Puberty may, thus, be a time when children, particularly girls, show an increased vulnerability to the emergence of psychosocial problems as a result of incomplete cortico-limbic development resulting from psychosocial stress exposure. In addition, some individuals are genetically predisposed to respond differentially to stress, as individuals with a specific polymorphism in the gene encoding the serotonin (5HT) reuptake transporter (SERT) are more susceptible to stressors. We propose that psychosocial stress interacts with genetic vulnerability to induce dysregulation of the LHPA axis to disrupt puberty and delay exposure to increases in E2, thus placing these females at risk for neurobiological defects and mood disorders. This project will study the behavioral, physiological, and neurobiological consequences of psychosocial stress, imposed by social subordination, during adolescence in female rhesus monkeys. Specific Aim 1 will test the hypothesis that social subordination, exacerbated by the presence of the short allele in the SERT gene, produces LHPA dysregulation and delays puberty. Aim 2 will test the hypothesis that exposure to psychosocial stressors during adolescence increases emotional reactivity by prolonging the pubertal transition and reducing exposure to E2, particularly in females with the short allele in the SERT gene. Aim 3 will use neuroimaging to test the hypothesis that development of cortico-limbic circuits and 5HT systems regulating emotional behavior are adversely affected by exposure to psychosocial stressors and reduced levels of E2. Aim 4 will test the hypothesis that SSRI therapy to subordinate females may normalize LHPA activity but not growth or puberty, delaying exposure to increasing levels of E2, and attenuating maturation of cortico-limbic circuits and 5HT systems. These studies will elucidate the complex interplay between behavior, genetics, and reproductive maturation, providing a comprehensive understanding of the role of adolescence as a critical period for the emergence of mood disorders in girls. PUBLIC HEALTH RELEVANCE: Using a rhesus monkey model, this project is designed to provide a better understanding of how psychosocial stress, imposed by social subordination, affects brain maturations and emotional development in females and whether this is influenced by polymorphisms in the gene that encodes the serotonin re-uptake transporter (SERT), a protein essential for normal serotonin neurotransmission and emotionality. These studies will elucidate the complex interplay between behavior, genetics, and reproductive maturation, providing a comprehensive understanding of how adolescence represents a critical period for the emergence of psychiatric disorders.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Socio-environmental factors significantly affect the risk for developing a number of behavioral and psychiatric disorders in people, including anxiety, depression, eating disorders, and addiction. It is becoming increasingly clear that some individuals are more susceptible to developing these pathologies and that this increased risk may be attributed to genetic factors. While identifying genetic/environment influences on behavior presents practical challenges and ethical constraints in people, rhesus monkeys represent an ideal model to begin to identify the genetic contribution to such disorders. The goal of this project is to conduct an analysis of the genetic heritabilities of a number of biobehavioral and physiological traits that are relevant for psychiatric disorders, followed by linkage and association analysis of candidate genes and gene polymorphisms that account for individual differences in these characteristics using the pedigreed rhesus monkey population living in large social groups at the Field Station. In the past year we collected phenotypic data on the animals. These data included behavioral measures of sociality and aggression, anthropometric measures of body weight, height, BMI, and serum hormone measures of leptin, insulin, triiodothyronine, thyroxin, glucose, and cortisol;and CSF measures of monoamines, CRH, vasopressin, and oxytocin. Data are being analyzed. This project represents a unique opportunity to conduct a translational, behavioral genetics study of candidate genes that predispose individuals to the adverse consequences associated with behavioral and psychiatric disorders, including anxiety and depression.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This project is designed to develop to collect pilot data in support of a grant application to NIH to better understand how changes in monoamine systems emerge during pregnancy that may predict what females may be more likely to exhibit postpartum depression symptomotology. Using microPET neuroimaging, the pilot study showed that dopamine 2 (D2R) receptor binding changed from pregnancy to the early postpartum interval. Because differences in D2R has been implicated in depressive symptoms these data that a rhesus monkey model can be used to show how D2R plasticity during the peripartum period is associated with socio-emotional behavior changes.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Using a rhesus monkey model, this project is designed to provide a better understanding of how psychosocial stress, imposed by social subordination, affects brain maturations and emotional development in females and whether this is influenced by polymorphisms in the gene that encodes the serotonin re-uptake transporter (5HTT), a protein essential for normal serotonin neurotransmission and emotionality. In this initial of the project, the first cohort of animals were recruited. These animals are embedded in large social groups at the Field Station and are distributed throughout the social status hierarchy of each group. In addition, 19 animals have the l/l 5HTT genotype and 20 animals have the l/s or s/s 5HTT genotype. Studies began this year included a dexamethasone suppression test to assess glucocorticoid negative feedback;measures of activity using a device (Actical) worn by the animal;tests of emotionality using the approach/avoidance test;and neuroimaging of the serotonin (5HT) 1A receptor using PET. These studies will elucidate the complex interplay between behavior, genetics, and reproductive maturation, providing a comprehensive understanding of how adolescence represents a critical period for the emergence of psychiatric disorders.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Gestational diabetes mellitus (GDM) is an increasingly prevalent problem with important health and societal impact, yet controversies continue in the most elementary issues related to GDM. While clinical studies are assessing the effects of maternal hyperglycemia on subsequent development of obesity, insulin resistance, diabetes, and cardiovascular disease risk in the offspring, progress is hampered by the absence of a good experimental animal model. While clinicians report that frank gestational diabetes is rare in the monkey population at Yerkes, they are presented with only the most advanced cases. No systematic studies of glucose tolerance have been done in the captive population. During the year, we initiated a pilot project to identify pregnant female rhesus monkeys at risk for gestational diabetes. Of 10 females studied during pregnancy, three have serum glucose in excess of 160 mg/dl, clearly indicative of potential problems with glucose regulation. Data are being collected on a large number of rhesus females to better understand the incidence in our population The data generated in these studies will serve as the nucleus for grants submitted to the NIH (e.g. NICHD, NIDDK, NHLBI) and foundations, such as the American Diabetes Association and March of Dimes. The rhesus monkey model blends well with our concurrent study of human gestational diabetes and a two-species approach will provide a unique edge in national competitions.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This is a translational multi-center project to initiate novel studies into the pathophysiological origins of a highly prevalent reproductive and metabolic disorder in women, namely polycystic ovary syndrome (PCOS). This hyperandrogenic syndrome increases a woman's lifetime risk of type 2 diabetes, cardiovascular disease, endometrial cancer and infertility. Our focus on PCOS stems from the highly pervasive, but heterogeneous nature of the disease, and our incomplete understanding of its etiology in women. We now need a naturally-occurring phenotype in a nonhuman primate model as well as an ability to target children at risk of PCOS to develop novel clinical interventions to prevent or diminish the adult PCOS phenotype. In this pilot project begun in late Fall 2009, we are establishing the efficacy of both nonhuman primate and human approaches with a consortium of multiple Primate Centers and the Wisconsin CTSA. The PI is David Abbott at the WNPRC. Our targeted use of appropriate nonhuman primate populations and expertise at three Primate Centers minimizes redundancies in resources available nationally and our utilization of Assay Services within the WCTSA demonstrates our sharing of a single core resource. The Yerkes investigators have completed sample collection on 150 adult female rhesus monkeys. All samples have been shipped to Dr Abbott at the WNPRC for analysis.

DESCRIPTION (provided by applicant): The social environment has a clear and profound impact on human health and well being. Chronic social stress and reduced access to social support are predictive of a number of adverse health outcomes, including cardiovascular disease and diabetes. Indeed, evidence suggests that social stress is linked to life expectancy itself: poor social integration, for example, has been estimated as a risk factor for mortality on the scale of familiar health risks like smoking and obesity. However, despite keen interest in social stress as a human health concern, the mechanistic relationships linking social stress to its impact on the body are still poorly understood, particularly on the level of the genome. The goal of the proposed work is to address this gap by investigating how dominance rank in female rhesus macaques influences genome regulation. Dominance status in macaques is an excellent model for human social stress: the natural hierarchical organization of macaque social groups is characterized by increased rates of harassment and threats directed towards lower ranking group members, which are reflected in rank-related stress physiology. Additionally, dominance rank assignments can be experimentally imposed in this species by altering group membership. Thus, an individual's exposure to social stress can also be manipulated, yielding an experimental system for investigating the effects of social stress on the genome that is directly translatable to humans, but that is practically and ethically impossible in humans themselves. The proposed study will take advantage of this system to investigate how dominance status and differential exposure to social stress influence gene expression in immune cells in the peripheral blood. This relationship will be investigated in four different cell types that play unique roles i the immune system, as well as under both day-to-day conditions and in response to stimulation by compounds that mimic disease infection. The project will also investigate the contribution of DNA methylation, an important mark that changes the structure of DNA without altering its sequence and that is known to respond to environmental context, to social status- related gene expression variation. Importantly, the study design will include a mid-study intervention to systematically alter the dominance rank positions of each individual in the study. This approach both permits the identification of genes that are causally influenced by exposure to social stress, and also permits investigation of whether an individual's social stress history has a long-term impact on the genome even after this stress has been alleviated. Together, these efforts will illustrate how social stress changes gene expression in an important animal model for human social stress, including how stress mitigation might offset the physiological costs of prior stress and how increased stress may alter individual vulnerability to disease.

DESCRIPTION (provided by applicant): The development of effective strategies to prevent and treat HIV infection is of paramount importance to improve global human health. Simian immunodeficiency virus (SIV) infection of rhesus macaques (Macaca mulatta) is the most effective pre-clinical model of HIV infection. Members of the Macaca genus, however, harbor several viruses that could confound experimental results from studies relating to the AIDS prevention or cure. It is therefore vital to ensure a constant supply of specific pathogen free (SPF) rhesus macaques for experimental studies in AIDS research. Since 2002, the SPF Rhesus macaque breeding groups at Yerkes National Primate Research Center (YNPRC) have, in part, been supported by U24 funding. By the end of 2013, all breeding groups will be free of SIV, Simian T-Lymphotropic Virus (STLV), Simian Type D Retroviruses (SRV), and Herpes Simian B Virus (Herpes-B). The central aim of this final phase of U24 funding is to achieve financial sustainability for those U24-supported groups and to expand multi-generational, genetically characterized breeding groups to provide appropriately aged experimental animals for NIH funded AIDS research. To achieve this aim, proven management practices will be employed that allow optimum production of animals in socially stable breeding groups to provide animals to AIDS-related studies. A second aim of this project is to expand methodology for SRV and Herpes testing in the Yerkes Virology Core. On-site SRV and Herpes B testing will save costs and allow a more rapid colony management response should the virus status of any SPF animal be questionable. A third aim of this renewal is to transition to a more cost-effective, therefore financially sustainable, SNP-based and direct sequencing approach for macaque genotyping, providing AIDS investigators with pedigreed animals with confirmed sets of major histocompatibility complex (MHC). It is expected that the YNPRC SPF rhesus macaque breeding colony will be financially sustainable by the end of the next U24 funding period and will be recognized as a national resource of genetically well-characterized rhesus macaques available for HIV/AIDS research.

DESCRIPTION (provided by applicant): Studies of both animals and children show that postnatal stress may have lasting effects on brain structure and function, resulting in behavioral and cognitive impairments, particularly for females. It is also unclear how social stress experienced by the mother during gestation synergizes with postnatal stress experienced by her offspring to produce these phenotypes. Importantly, other environmental factors that may interact with stressor exposure to affect brain development during childhood are frequently overlooked, most notably the consumption of calorically dense diets (CDDs) and the resulting metabolic phenotype. Indeed, there is likely a synergy, as chronic social stress is a cumulative risk factor for childhood obesity. Not only may obesity accelerate the tempo of puberty but limited data in children suggest the developing brain is vulnerable to these metabolic insults, as increased body fat is associated with altered brain structure and deficits in cognition and emotional processing. Understanding the impact of stress and obesity on neurodevelopment is critically relevant, given alarming rates of obesity in children, likely due to the consumption of CDDs - a dietary environment quite unlike the typical low caloric diets fed animals used as models for children. Key biological signals could be stress-induced elevations in cortisol and proinflammatory cytokines that are exacerbated by increased fat mass. Prospective studies of the developmental origins of health and disease are difficult to do in children. However, socially housed rhesus monkeys provide an effective translational model, as social subordination produces distinct stress-related phenotypes even during development. This application will address four specific aims to test the overarching hypothesis that prenatal maternal stress interacts with post natal social stress to alter female neurobehavioral development from infancy through puberty and these impairments are exacerbated by obesity. Aim 1 will determine whether increased fat mass interacts with postnatal social stress to alter developmental trajectories of female social and emotional behavior, as well as prefrontal-related cognitive function. Using neuroimaging, Aim 2 will test the hypothesis that social stress and increased fat mass will synergize to alter structural and functional development of the prefrontal cortex (PFC) and its connectivity with regions regulating social and emotional behaviors as well as executive function and self-regulation from infancy, with differences accelerating through the pubertal transition. Mediation analysis in Aim 3 will examine whether cortisol and inflammatory markers mediate the effects of social stress and fat mass on impaired neurobehavioral development. Using cross-fostering, Aim 4 will determine how maternal stress during gestation synergizes with postnatal social stress and obesity to further compromise neurobehavioral development. The project will identify potential biological signals that mediate the adverse effects of stress ad obesity on brain health and behavior and, in doing so, will provide crucial information that will help shape clinical interventions and social policy improvement to optimize neurobehavioral development in girls.

DESCRIPTION (provided by applicant): Emotional feeding resulting from chronic exposure to psychosocial stressors is likely a key factor for excess food intake. This may be particularly important for women, who consistently report more stress-induced eating and have higher rates of obesity. The well-accepted notion is that emotional feeding of calorically dense diets is a form of self-medication to relieve the behavioral and physiological manifestations of stress. While this may be the case, it is also probable that emotional feeding is the result of compromised dopamine (DA) reward pathways, as exposure to chronic psychosocial stress increases susceptibility to an addictive phenotype by reducing DA 2 receptors (D2R) in mesolimbic regions, producing a hypodopaminergic condition. Furthermore, simply eating a calorically dense diet reduces D2R in these regions but only in some animals. Thus, self-medicating with high caloric diets may further compromise the stress-induced hypofunctional DA system. Clearly, this is not a healthy coping strategy for unresolved stress. An issue particularly important for women attempting to diet is whether these stress-induced changes, including impaired DA function, persist in the face of life style changes. Our preliminary data using social subordination in female rhesus monkeys as a model of chronic psychosocial stress in women suggests this may be the case, as subordinate females consume significantly more calories when given a high caloric diet (HCD) and this hyperphagia persists when a healthier diet is exclusively available. What is not known, however, is how this pattern continues under these healthy dietary conditions. Similarly, it is unknown if the alleviation of chronic psychosocial stress will restore caloric restraint, or if changes in systems regulating appetite, including DA, persist even after stress is resolved. Using socially housed female rhesus monkeys as a translational model for women, this project will identify mechanisms that sustain emotional feeding. Aim 1 will determine whether chronic social stress induced by social subordination sustains excessive intake of a HCD and whether this is exacerbated by exposure to acute stressors. Aim 2 will test the hypothesis that intake of a calorically dense diet will reduce DR2 availability in mesolimbic regions and this will be exacerbated by social subordination. Aim 3 will test the hypothesis that estradiol will be more effective suppressing caloric intake in dominant females when a LCD is available but will promote caloric intake when an HCd is available, particularly in subordinate females. in a Aim 4 will use two intervention strategies to further elucidate how social stress sustains emotional feeding: 1) despite replacing a HCD with a healthier low caloric diet, hyperphagia and reduced D2R binding potential in subordinates will persist; and 2) reducing social stress by changing social status will improve but not normalize D2R availability or food intake Together, these studies will increase our understanding of factors that sustain emotional feeding, even in a healthy dietary environment.

PROJECT SUMMARY In most mammalian species, social interactions among individuals of the same species are governed by dominance relationships. These hierarchical relationships are established and maintained by agonistic behaviors, including aggression. Importantly, recent data indicate that the neural mechanisms underlying aggression and attaining dominance produce a phenotype that is resistant to social stress while the mechanisms underlying subordinate status produce a social stress-susceptible phenotype that may result in a number of adverse behavioral and physiological outcomes. Despite the relationship between social status and stress, the neurochemical mechanisms that underlie dominance have received only limited attention in males and almost no attention in females. This project will fill this critical gap in our knowledge by testing an integrated series of hypotheses using Syrian hamsters and rhesus monkeys. This project will critically test the overarching hypothesis that the agonistic behaviors responsible for the formation and maintenance of dominance relationships are regulated in dramatically different ways by vasopressin (AVP) and serotonin (5- HT) in males and females. Specifically, we propose that activation of AVP and inhibition of 5-HT promotes dominant status and a stress resistant phenotype in MALES while producing subordinate status and a stress susceptible phenotype in FEMALES. In contrast, inhibition of AVP and activation of 5-HT promotes dominance and a stress resistant phenotype in FEMALES while producing subordinate status and a stress susceptible phenotype in MALES. Together, these data will significantly expand our knowledge of sex differences in the neurochemical mechanisms that define social phenotypes and will provide innovative gender specific strategies for promoting resistance to social stress. The data obtained in this project could have an almost immediate clinical impact by guiding drug treatments for stress reduction in men and women as well as guiding drug development by emphasizing the role of AVP-targeted drugs in males and 5-HT- targeted drugs in females.