Kim Wallen - US grants

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.

This is a request for an ADAMHA RSDA Level II award. The research investigates gender development in group-living male and female rhesus monkeys exposed to atypical levels of prenatal androgen. These studies will elucidate the role that prenatal androgens play in regulating genital and psychological differentiation in a primate, under social conditions that produce a full range of gender-related social and sexual behavior. By employing short-term treatments during early and late gestation the project seeks to separate the effects of prenatal androgen on masculinization of the genitalia from its effects upon masculinization and defeminization of behavior. These studies are relevant to issues of human gender differentiations and discordances between genital sex and gender role behavior, a clinical problem that affects a substantial number of humans. To achieve these goals, time-mated pregnant rhesus females, living in large age-graded heterosexual groups, will have the sex of their fetus identified using sex chromatin staining. Mothers of female fetuses will be injected with testosterone enanthate (TE) or vehicle for 35 days starting on gestational day 40 or 110. Mothers of male fetuses will be injected with a Nal-Lys GnRH antagonist (Antide), Antide plus TE, or vehicle starting at the same times and for the same duration as the female treatments. Prenatal Antide will suppress testicular function creating males who are androgen deficient for a brief period early or late in gestation. The times selected represent periods during or after genital masculinization and when previous work has shown androgens to differentially affect the sexually dimorphic patterns of rough play and juvenile mounting. Experimental and control animals will be observed behaviorally from birth through the transition into adulthood using methodology that captures traditional sexually dimorphic patterns of behavior and additional patterns of social behavior that are only observable when infants can interact with all ages and sexes in an unrestricted manner. Endocrine and skeletal development will be tracked with systematic collection of serum samples to measure neonatal and peripubertal patterns of hormone secretion and morphometric measures of skeletal growth. Pituitary function will be challenged with exogenous GnRH at several times during development to assess the effects of early androgen exposure on the integrity of the hypothalamo-pituitary-gonadal axis. After the pubertal transition, male and female sexual behavior will be investigated in response to their endogenous hormones. If experimental females produce offspring, the relationship between early androgen exposure and materna] behavior will be investigated. These studies provide the first experimental manipulation of prenatal androgen in male primates and will substantially extend our understanding of the role androgen plays in gender differentiation in primates. RSDA support will free the PI from teaching and administrative duties and allow the development of new behavioral and diagnostic techniques for assessing the effects of prenatal androgen manipulations.

DESCRIPTION (Adapted from applicant's abstract): This is a proposal for the continuation of a Research Scientist Development Award, now termed an Independent Scientist Award (KO2). During the first term of this award 60 experimental and control monkeys were created that had either normal or atypical prenatal exposure to androgen. Androgen exposures were of 40 days or less duration and critically timed to the start of the second or third trimester of pregnancy. Prenatal androgen exposure was reduced by twice daily administration of flutamide, an androgen receptor blocker. Weekly injections of testosterone enanthate were used to increase androgen. The proposed K02 Award will allow the PI to devote the majority of his time to investigating the behavioral, neuroendocrine, and cognitive consequences of these alterations in prenatal androgen exposure in socially-living rhesus monkeys. These studies will provide fundamental information about how early hormone exposure affects the development of sexual and sex-typed behavior in a primate with a complex social organization and a long period of sexual development. One of the most striking human cognitive sex differences is in aspects of spatial cognition. The proposed support will allow the PI to develop computerized approaches to studying spatial cognition in group-living monkeys to take advantage of a large sample of rhesus monkeys with atypical androgen exposure created during the previous funding period. In addition, the increased time available for research will allow investigation of possible sex differences in brain structure using magnetic resonance imaging. If neuroanatomical sex differences are demonstrated they can then be pursued in the experimental subjects as they near adult development. During the term of this award the experimental and control subjects will go through puberty and enter adulthood. A goal of the proposed plan is to explore how alterations in the prenatal androgen environment affect the process of pubertal change and adult reproductive behavior. Because none of our hormonal treatments substantially altered the genitalia of experimental females there are no apparent physical barriers to pregnancy. It will be particularly interesting to determine whether substantially less drastic alterations in prenatal androgen exposure than those previously used produce marked changes in reproductive and maternal behavior. In males that have experienced reduced prenatal androgen exposure the PI will determine whether the timing of such hormonal alterations differentially affects their sexual behavior. These studies of nonhuman primates have particular relevance to issues of human gender development, particularly the occurrence of gender dysphoria. The last goal of the proposal is to allow the PI to gain a deeper understanding of human gender dysphoria by spending time at the Clarke Institute of Psychiatry in Toronto Canada, which has one of the world's most active research and treatment programs for childhood and adult gender dysphoria. The combination of these efforts will allow the PI to expand his research into new areas, while increasing his understanding of its relevance to important human mental health conditions.

The effect of altering the prenatal hormonal environment on behavioral development and neuroendocrine function was studied in genetic males and females exposed to either and androgen receptor blocker (flutamide), an long-lasting androgen (testosterone cypionate), or vehicle controls. Treatments were given at the beginning of the second (early) or third trimester (late). Only early flutamide treatment produced substantial genital modification to treated subjects. Early androgen treatment reduced neonatal responsiveness to exogenous gonadotropin-releasing hormone (GnRH) in females, while late flutamide treatment increased responsiveness and basal LH secretion. Behavioral and neuroendocrine development of these unique monkeys will continue through puberty and into adulthood. This work provides fundamental information about the role prenatal androgenic hormones play in the sexual differentiation of behavior and endocrine function and is relevant to understanding in humans how small modifications in the early hormonal environment affect later development.

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 early hormone environment has been thought to influence the development of male/female behavioral and neuroendocrine differences through the life span. This study exposed developing male and female rhesus monkeys to atypical prenatal hormonal environments and has been investigating their behavioral and neuroendocrine development. Treatments were designed to mimic accidental exposure to or suppression of androgens prenatally. This study investigated whether prenatal androgen exposure, social rank, and body weight are factors regulating pubertal development in female rhesus monkeys. Subjects'mothers received injections of testosterone enanthate (20 mg/week), flutamide (an androgen receptor blocker, 30 mg/kg twice daily), or vehicle during gestational days 35/40-70 (early) or days 105/110-140 (late). We investigated whether reducing endogenous T prenatally affected adult copulatory behavior in male rhesus monkeys. Treatment with flutamide early in gestation resulted in males with significantly less masculinized genitals that control males or males getting flutamide late in gestation. During the current year all of the prenatally treated males and females along with concurrent controls were MRI scanned to look for sex differences in brain anatomy. In collaboration with investigators at the National Institute of Aging control MRI scans of males and females are being evaluated for structural sex differences. The prenatally treated animals are then being compared to the common morphed brains of control males and females to identify where prenatal treatments have modified brain structures.

We propose continued studies of the life-span development of 64 male and female rhesus monkeys exposed to normal or atypical levels of prenatal androgen. Treatments were 35-40 days long and timed to the start of the second or third of gestation. Endogenous prenatal androgen effects were reduced by twice daily administration of flutamide, and androgen-receptor blocker. Weekly injections of testosterone enanthate increased androgen. Continuation of this project will allow investigation of the behavioral, neuroendocrine, and cognitive consequences of alterations in prenatal androgen exposure in socially-living rhesus monkeys during the pubertal transition and as adults. These studies provide fundamental information about the effects of prenatal androgens on development of sexual an sex-typed behavior in a primate with a complex social organization and extended sexual development. Subjects will go through puberty and enter adulthood during this funding period. We plan to explore how altered prenatal androgens affect pubertal change and adult reproductive behavior. Because none of our hormonal treatments altered the genitalia of experimental females in a manner to prevent pregnancy we can explore whether exposure to much lower levels of prenatal androgen than previously studied after female reproductive and maternal behavior. In addition to studied of reproductive behavior and neuroendocrine function we will investigate differences in vocal communication, otoacoustic emissions (OAEs), cognition, and neural sex differences. Males and females differ in vocal repertoires. We will investigate the development of these sex differences and relate them to differences in prenatal hormone exposure. Spontaneous and click-evoked OAEs offer a noninavise window into neural function and differ between male and female humans. We will investigate this sex difference in rhesus monkeys and relate it to our atypical prenatal androgen treatments. The most striking human cognitive sex differences is in spatial cognition. We will study sex differences in spatial cognition in group-living monkeys and investigate the relationship between spatial cognition and atypical prenatal androgen exposure. Exploratory studies will relate behavior differences to noninvasively imaged difference in brain structure. We are exploring the hypothesis that the timing of altered prenatal androgens differentially affects sexual behavior, anatomical, neuroendocrine function, and cognition. Particularly significant is our finding that androgen manipulations can affect behavioral and neuroendocrine development without any anatomical changes. These nonhuman primate studies are relevant to human gender development both normative individual variation and the occurrence of gender dysphoria. The range of effects we will investigate reflect the effects of altered hormonal conditions that could presumably occur in humans from maternal illness or exposure to environmental toxins. This research extends and develops the notion that alterations in the prenatal androgen that leave no external indicators may produce life-long alterations in development and adult behavior.

Most of the axial growth of the eye occurs during the first two years of life. While the growth of the eye is usually precisely regulated by a process known as emmetropization, this process can become deranged in certain conditions resulting in excessive or retarded axial growth and large refractive errors. This project seeks to elucidate the molecular mechanisms responsible for the physiological processes that cause aberrant ocular growth with aphakia. Specifically, we will assay infantile eyes for changes in certain neurotransmitters and growth factors after removing the crystalline lens which we have previously shown retards ocular growth. Understanding the mechanisms whereby aphakia alters ocular growth is of vital importance to accurately select the most appropriate intraocular lens power to implant into infantile eyes after cataract surgery. In addition, by understanding the mechanisms retarding ocular growth, it may be possible to modulate the excessive ocular gro wth which occurs in certain pathological conditions such as retinopathy of prematurity. FUNDING NIH EY08544 $156,904 9/30/98 - 1/31/02 PUBLICATIONS None P51RR00165-38 1/1/98 - 12/31/98 Yerkes Regional Primate Research Center

DESCRIPTION (provided by applicant): The hypothesis is investigated that female sexual interest is stimulated by the neural actions of ovarian estrogens and that androgens regulate the bioavailability of these estrogens through interactions with sex hormone binding globulin (SHBG). Three projects, using a rhesus monkey model of endocrine function and behavior, investigate the hormonal basis of female sexual initiation. Project I investigates sexual initiation in females across the menstrual cycle, comparing the occurrence of female sexual initiation in a social group context during normal cycles treated with an androgen receptor blocker (flutamide) or an estrogen receptor blocker (tamoxifen). This will clarify whether androgens or estrogens act neurally to modulate female sexual motivation. Project II tests the novel hypothesis that SHBG regulates bioavailable estrogens and androgens through these steroids' different affinities for SHBG. This project uses a monkey model of hormonal replacement therapy for reproductively prime females after surgical removal of their ovaries and tests the hypothesis that chronic estradiol (E2) ceases to effectively stimulate female sexual interest as estrogen is sequestered by SHBG. It further investigates whether an androgen, 5a-dihydrotestosterone (DHT), with a markedly higher affinity for SHBG than estradiol can acutely and rapidly reinstate female sexual interest by increasing free estradiol by displacing SHBG-bound estradiol. Ovariectomized females receiving chronic estradiol treatment mimicking mid-follicular estradiol levels will be observed for sexual initiation during chronic E2 treatment alone and following chronic E2 and an injection of DHT or E2. Concurrent administration of flutamide or tamoxifen with the estrogen or DHT will discriminate between behavioral changes resulting from the activation of neural androgen or estrogen receptors. The effects of these treatments on neuroendocrine function will also be investigated. Project III investigates whether common human hormonal replacement therapies of chronic estrogen, or chronic estrogen plus testosterone with or without concurrent progestin, can reinstate female sexual interest in reproductively prime ovariectomized female monkeys. The hypothesis will be tested that chronic progestin therapy reduces or eliminates the effectiveness of therapies that reinstate female sexual interest without progestin. These therapies will also be compared on their effects on neuroendocrine function. These studies will markedly increase our understanding of the role that ovarian steroids play in modulating women's sexuality.

DESCRIPTION (provided by applicant): Understanding the relationship between psychopathology and basic psychological and biological processes is crucial to curing significant mental health problems. We propose a program training clinical psychologists in basic science related to psychopathology and basic scientists in the nature of psychopathology. Focusing on how neuroendocrinolqgy and cognition relate to the development and expression of psychopathology, 9 predoctoral fellows will receive a coordinated three year training program starting in their 3rd year of graduate study. Selected from a pool of over 50 potential candidates already enrolled in four participating graduate programs, the fellows will be a unique cohort of students interested in bridging basic and applied science to understand psychopathology. The 24 participating faculty come from the Department of Psychology of Emory College, Psychiatry and Behavioral Sciences, and the Yerkes National Primate Research Center. The training program involves four graduate programs: Clinical Psychology, Cognition and Development, Neuroscience and Animal Behavior, and the Neuroscience Training Program of the Graduate Division of Biological and Biomedical Sciences. The faculty's expertise 'covers psychopathologies from autism to schizophrenia and provides a unique training environment focusing on both human and nonhuman primates, as well as other animal models. Trainees have a mentor and a co-mentor from different graduate programs, fostering collaboration and increasing training diversity. Trainees will have completed the basic course work required by their graduate programs prior to program entry allowing focus on the training program offered by this fellowship. Trainees participate in a two-semester psychopathology sequence taught by clinical and basic science faculty exploring specific psychopathologies and relevant animal models. The first semester offers a high-level introduction to human psychopathology, assessment, methodologies, and treatment. The second semester focuses on the development of, and neuroendocrine and cognitive influences on, five psychopathologies: anxiety, depression, schizophrenia, developmental disorders, and personality disorders with consideration of relevant animal models. Trainees will acquire first hand knowledge of the nature of the psychopathology in addition to relevant basic research background. In addition to this core sequence in psychopathology, trainees take two additional courses, one in neuroscience and one in behavioral/cognitive science, developing their basic science skills in areas relevant to psychopathology. This training program will not provide clinical training, instead develops clinical researchers who are comfortable with and trained in the psychobiological systems underlying psychopathology and basic researchers who are comfortable with and trained in human psychopathology. Clinical trainees will get the opportunity to delve more deeply into the basic science underlying psychopathology while basic science trainees will come to understand the relationship between basic science and human mental illness.

[unreadable] DESCRIPTION (provided by applicant): The amygdala, a sexually differentiated brain structure intimately involved in emotions, cognition, social behavior, and sexuality, is particularly important in monkeys for recognizing and responding to social context and danger. This structure continues to develop postnatally and is sexually differentiated prior to puberty, yet nothing is known about its contribution to sex differences in behavior and cognition, it impact on neuroendocrine function or its role in modulating social signals influencing puberty. Puberty in monkeys and humans is affected by social context and the amygdala may be one of the modulators of the interaction between social context and neuroendocrine function. We will investigate social, emotional, cognitive and neuroendocrine consequences of neonatal and post-pubertal amygdalectomy in male and female rhesus monkeys. Subjects will live in semi-natural complex social groups. A team with expertise in emotional regulation, cognition, neuroendocrine function, and social behavior, will track the development of these systems and relate changes in juvenile function to the pubertal transition and adult social behavior. Forty- eight monkeys, distributed between 8 male and 8 female controls, 8 neonatally amygdalectomized of each sex, and 8 post-pubertally lesioned subjects of each sex, will be created in two annual cohorts of 24. Neonatal gonadal function, anxiety, fear, and maternal attachment will be assessed along with extensive observations of social interactions in the subjects' natal group during the first year of life. Social observations, assessment of emotionality, and stress physiology will continue for the 2nd through 4th years of life. At two years of age, cognitive tasks, using continuously available computer kiosks, will assess object and spatial memory span and object discrimination. Sex differences in social interactions and social integration will be investigated through detailed observations of social behavior. At two years of age females will start intense endocrine sampling to assess pubertal timing as well as pubertal changes in social and sexual behavior. Three year old males will be removed from their natal group to simulate male migration and formed into bachelor groups for investigation of the pubertal transition. It is hypothesized that the different male and female natal social environments, combined with the sexually differentiated nature of amygdala function will result in different magnitudes of effect of amygdala damage, with males likely more severely affected than females. We hypothesize that amygdalectomized females will uniformly show early puberty. Altered amygdala function is found in a number of neurodevelopmental disorders and while amygdalectomy does not serve as a model for any single human mental disorder, understanding its functional effects on a range of behavioral, cognitive and neuroendocrine endpoints will be invaluable for understanding the developmental trajectory of alterations produced by early amygdala dysfunction in human neurodevelopmental disorders. [unreadable] [unreadable] [unreadable]

21+ years old; Adolescent; Adolescent Youth; Adult; Affect; Age; Amygdala; Amygdaloid Body; Amygdaloid Nucleus; Amygdaloid structure; Anxiety; Behavior; Behavioral; CRISP; Class; Cognition; Cognitive; Cognitive Discrimination; Complex; Computer Retrieval of Information on Scientific Projects Database; Computer Systems Development; Computers; Condition; Development; Development, Computer Systems; Discrimination; Discrimination (Psychology); Dysfunction; ENPT; Emotional; Emotional disorder; End Point; EndPointCode; Endpoints; Fear; Female; Fright; Functional disorder; Funding; Grant; Human; Human, Adult; Human, General; Institution; Investigators; Lesion; Life; Macaca mulatta; Man (Taxonomy); Man, Modern; Memory; Modeling; Monkeys; NIH; National Institutes of Health; National Institutes of Health (U.S.); Nature; Neonatal; Neurodevelopmental Disorder; Neuroendocrine; Neuroendocrine System; Neurological Development Disorder; Neurosecretory Systems; Numbers; Physiopathology; Psyche structure; Puberty; Range; Research; Research Personnel; Research Resources; Researchers; Resources; Rhesus; Rhesus Macaque; Rhesus Monkey; Sampling; Sex Behavior; Sex Characteristics; Sex Differences; Sexual Activity; Sexual Behavior; Social Behavior; Social Conditions; Social Environment; Social Interaction; Societal Conditions; Source; Systems Development; Time; United States National Institutes of Health; adult human (21+); amygdaloid nuclear complex; cohort; design; designing; gender difference; gonad function; gonadal function; human puberty; juvenile; juvenile human; male; mental; pathophysiology; sex; sex activity; sexual dimorphism (noncellular); social; social climate; social context; social group; sociobehavior; sociobehavioral; socioenvironment

DESCRIPTION (provided by applicant): There is an unmet need for new and safer estrogen therapies (ET) to treat the symptoms associated with low levels or a lack of estrogens, including decreased sexual motivation, hot flushes, sleep disorders, depression/anxiety, brain and spinal cord trauma, amyotrophic lateral sclerosis, and cognitive dysfunction. Currently only estrogens alone or in combination with progestins or androgens are available to treat these conditions, including natural or surgical menopause. However, many women cannot or do not want to take estrogens because of their potential side effects, the most critical of which are the effects of estrogen on the uterus and the breast. However, estrogen insufficiency contributes to post-menopausal decline in sexual desire that can lead to female sexual desire disorder (FSDD). Current hormonal therapies for FSDD use treatment with an estrogen and testosterone, but their effectiveness is limited. Estradiol by itself can reinstate sexual desire if given at dosages that produce midfollicular serum estradiol levels (100-300pg/ml). However, at such dosages there are concerns about the risk of breast or uterine cancer, thus all current FSDD treatments use hypoestrogenic treatment (20pg/ml) in conjunction with testosterone when larger amounts of estradiol would likely increase treatment effectiveness. Here we investigate a novel and potentially safe estrogen therapy using 10b,17b-dihydroxyestra 1,4 dien-3-one (DHED) to reinstate sexual receptivity and sexual motivation in rat and monkey models. DHED is an inactive precursor of estradiol; it does not bind to estradiol receptors and thus produces no estrogenic effects in its native form. However, an enzyme converts DHED to estradiol. Our preliminary evidence suggests that this enzyme is only expressed in the CNS thus potentially making DHED a brain-selective prodrug for 17b-estradiol (E2). DHED, which crosses the blood brain barrier, would be inactive outside of the brain, but would deliver effective amounts of E2 to the brain. We propose to use sexual behavior as a model estrogen-responsive system with which to test DHED's potential therapeutic effects. Induction of lordosis in ovariectomized females rats using a sequential DHED and progesterone treatment will test the central activity of DHED. We will then use an ovariectomized monkey model of female sexual initiation to test DHED's efficacy in increasing sexual desire. Dr. Prokai will synthesize DHED for this project and undertake in vivo studies on conversion of DHED to E2. Dr. Merchenthaler's lab will investigate estradiol induced gene expression in the brain and periphery after acute or chronic DHED treatment and investigate DHED's capacity to active female rat sexual receptivity. Dr. Wallen's lab will undertake studies of female rhesus monkey sexual initiation during DHED treatment. Preliminary data from rodent models of hypoestrogenic conditions show that DHED is a brain-selective bioprecursor produg for E2, and is likely to be a safe and effective treatment of conditions due to the lack of estrogens (ovariectomy) or low levels of estrogens (menopause). If successful these studies will revolutionize the treatment of clinical problems resulting from hypoestrogenic conditions.

? DESCRIPTION (provided by applicant): Delineating patterns of social affiliation and hierarchical relations in group living rhesus monkeys is very labor intensive. We propose to develop and implement innovative new software and hardware technology that will automate the continuous collection of social interaction data in a complex species-typical rhesus monkey social group. The proposed system will continuously and simultaneously track the precise 3D locations of multiple individuals in a large social group of monkeys in their half acre living area Individual identification will come from comfortable collars, we will design and build, that contai active RFID tags that continuously broadcast each animal's unique code. RFID detectors within and around the compound's periphery allow triangulating each animal's precise location to within 25cm as well as their direction and velocity of movement. We will track the adult males and females that make up the core of the social hierarchy (45 individuals in total). Distance between individuals is a key diagnostic for inferring social affiliation as monkeys are rarely withn 25cm of each other by accident. Thus knowing the amount of time spent at that distance allows inferring affiliative relations. The patterns of affiliation, avoidance, and time in close proximit will allow inferring the social hierarchy of the group. In combination with social network analysis, we will identify patterns of group association and social instability. In addition, we will develop an deploy (in years 3 and 4) a RFID-guided camera system that automatically records high resolution video of predetermined target subjects without human intervention, allowing significantly increased efficiency for later human behavioral coding from these video snippets. Three software systems will be developed under this project: 1) behavioral inference software that infers affiliative, hierarchical, aggressive, and sexual behavior from the automated tracking data; 2) social network analysis software to identify social instabilities; 3) video recognition software that will automate the coding of social behavior from the video snippets collected with the RFID-guided cameras. Together this combined hardware and software will provide an enhanced animal resource making possible easy investigation of the social impact of a variety of manipulations, from drugs to neural insult. In addition, this system can be used to monitor group health and identify individual sickness behavior. This animal resource would facilitate research funded by NIMH, NIDA, or NICHD, and support and enhance primate colony management.