Linda P. Spear - US grants

Maturation of a neurotransmitter system may not be an "all or none" event; early maturing neurotransmitter systems or subsystems could serve to mediate age-specific behaviors of the neonate independently of their function in adults. The behaviors induced by psychoactive drugs early in development often are different from those induced by the same drug in adult animals and frequently mimic environmentally-induced behaviors seen at particular stages of ontogeny. For example, potent sensory stimuli (such as novel odors, milk when deprived, foot shock) and monoamine agonists induce marked behavioral activation in rat pups during the first postnatal week, while catecholamine agonists and potent sensory stimuli induce wall climbing behavior during the second postnatal week and rarely if ever thereafter. The work outlined in this proposal will determine which neurotransmitter system(s) mediate these early age-specific behavioral responses to potent sensory stimuli and psychoactive drugs during these two specific ontogenetic periods. Different sensory stimuli and drugs will be tested for their ability to induce behaviors such as general activation and wall climbing during the first and second week of life. Then, the efficacy of different neurotransmitter antagonists in blocking both the drug- and sensory stimulus-induced behaviors will be studied to determine which neurotransmitter systems(s) are critical for the expression of these induced patterns of behavior. This psychopharmacological approach will be supplemented by analysis through brain transections, electrolytic and neurotoxin lesions, and neurochemistry, to specify which brain regions are most critical ontogenetically for these behaviors. The convergence of information gathered through these various approaches should define more clearly the role of specific neurotransmitter systems or subsystems in the behavior of the developing animal.

The work outlined in this proposal will examine further the neurobehavioral consequences of gestational exposure to cocaine administered subcutaneously (s.c.) in Sprague-Dawley rat pups. In work conducted during the previous funding period, psychopharmacological and behavioral evidence was obtained consistent with the hypothesis that gestational cocaine exposure may result in an attenuation in dopamine (DA) activity, at least during the preweaning period. Young offspring exposed gestationally to cocaine were also observed to exhibit cognitive deficits in some but not all conditioning situations. The research outlined in this proposal will investigate further these apparent alterations in DA function and cognitive performance seen early in life following gestational cocaine exposure in Sprague-Dawley rate. Gavid dams will be s.c. injected daily with 10, 20 94 40 mg/kg/3cc cocaine hydrochloride from gestational day 8 (E8) to E20. Control groups include dams similarly injected with 0.9% saline and pair-fed to the 40 mg/kg cocaine dams, as well as non-treated dams. Offspring from all groups will be fostered to surrogate dams on postnatal day 1. In Specific Aim 1, DA function will be examined in offspring from these prenatal treatment groups throughout ontogeny in terms of assessment of DA metabolism (as an index of DA turnover), responsiveness to pharmacological challenges with apomorphine and haloperidol (using psychopharmacological, neurochemical and hormonal response measures), and examination of DA autoreceptor function. In Specific Aim 2, studies will examine the circumstances under which cognitive deficits are observed in neonatal, infant, weanling, periadolescent and adult offspring exposed gestationally to cocaine to determine whether observed deficits in conditioning and retention are related to degree of training, the sensory modalities used for conditioned stimuli, the type of unconditioned stimulus used for conditioning, and the type of response measure used for assessment of performance. Studies will also be conducted to assess whether disruptions in cognitive performance seen early in life are related to maturational delays in cognitive development, or to alterations in cognitive function per se. Such animal research examining the neurobehavioral consequences of early cocaine exposure is particularly important at this time, given the recent escalation in the number of human offspring exposed gestationally to cocaine.

cocaine; dopamine receptor; animal developmental psychology; embryo /fetus drug adverse effect; cognition disorders; dopamine; neurotransmitter metabolism; stimulus /response; psychopharmacology; neuroendocrine system; neurochemistry; memory; conditioning; weanling animal; mature animal; juvenile animal; newborn animals; infant animal;

The work outlined in this proposal is in response to a Request for Applications entitled "Biomedical and Behavioral Research on Alcohol and Youth". Despite the fact that alcohol use typically begins in adolescent or even younger youth, basic research examining the ontogeny of ethanol sensitivity in laboratory animals is relatively sparse and what data are available are surprisingly inconsistent. The work outlined in this proposal is designed as a straight-forward and systematic examination of the ontogeny of ethanol sensitivity, using well-established procedures used in developmental psychopharmacology and with consideration for the special challenges in working with ethanol. Animals of a variety of ages will be examined, including the periadolescent period, an age that we previously have shown to be associated with a variety of age-specific behavioral propensities, including an altered psychopharmacological sensitivity to a variety of drugs. The specific aims of this work are to characterize systematically the ontogeny of ethanol responsiveness in terms of acute ethanol sensitivity and with respect to the development of acute and chronic tolerance to ethanol. To accomplish these aims, three series of experiments are proposed. Experimental Series 1 will examine the ontogeny of acute ethanol sensitivity using a variety of response measures, including assessment of ethanol-induced hypothermia and suppression of startle as well as ethanol-induced alterations in locomotion and in swim speed; dose-response relationships will be systematically assessed, and blood and brain alcohol levels will be examined. Experimental Series 2 will assess the ontogeny of acute ethanol tolerance through the use of linear regression analyses of data on blood and brain alcohol levels at the time of recovery of the righting reflex in animals exposed to a variety of hypnotic doses of ethanol. Experimental Series 3 will examine the ontogeny of chronic tolerance to ethanol as well as the impact of early ethanol exposure on the later development of ethanol tolerance in adulthood; tolerance will be assessed in terms of the hypnotic effects of ethanol as well as ethanol-induced alterations in body temperature, startle amplitude and locomotor activity, with alterations in ethanol elimination rates being used as an index of the development of dispositional tolerance. The results of the studies proposed will provide a solid data base of fundamental information regarding the ontogeny of ethanol responsiveness, findings that may be of substantial importance given the early initiation of alcohol use among youth in the United States.

Adolescents differ from adults in responsivity to a number of ethanol effects. Their insensitivity to the motor incapacitating and sedating effects of ethanol may increase consumption capacities and support higher levels of adolescent drinking, thereby contributing to the unique risks associated with alcohol drinking during adolescence. It has yet to be established whether this unique pattern of alcohol responsivity during adolescence is a function of age-related differences in initial neural sensitivity to alcohol or in the magnitude of tolerance developing within ethanol exposure periods (acute tolerance) or across exposures (rapid and chronic tolerance). There are, however, recent observations of notable ontogenetic alterations in the expression of these ethanol adaptations. Such ontogenetic differences in ethanol tolerance may be particularly pronounced under stressful circumstances, given evidence that vulnerability to stressors may be increased during adolescence and that development of ethanol tolerance is enhanced by stressors. Consequently, the work outlined in this proposal will explore the contribution of tolerance to the unique pattern of alcohol responsivity during adolescence, and the effects of stressors on these adaptational processes. Studies will compare the expression of acute, rapid and chronic tolerance to various characteristic effects of ethanol and the impact of stressors on these ethanol adaptations in male and female, adolescent and adult (as well as weanlings, where feasible) Sprague-Dawley rats. It is predicted that age-related attenuations in sensitivity to specific ethanol effects will be associated with greater acute tolerance but less rapid and chronic tolerance to those effects, with adolescents being particularly vulnerable to stress-induced facilitation of acute, rapid and chronic tolerance. Characterizing the ontogeny of ethanol tolerance and the effects of stressors on these adaptational processes is not only critical for assessing determinants of the unique pattern of alcohol responsivity seen during adolescence, but also may contribute to our understanding of the long-term consequences of adolescent alcohol exposure for later alcohol use and abuse.

DESCRIPTION: (Adapted from the Investigator's Abstract) Alcohol use typically begins during adolescence in humans, with the age of initial use strongly predicting later alcohol abuse. Adolescents of a variety of species, including humans, differ neurobehaviorally from their adult counterparts; characteristic traits include an altered responsiveness to stressors as well as alterations in brain regions sensitive to activation by stressors and implicated in mediating the reinforcing effects of alcohol and other drugs of abuse. Yet, despite frequent alcohol use by adolescents and the implications of the early use for later abuse, study of alcohol and adolescence is still in its infancy. The work outlined in this proposal will use an animal model to examine the reinforcing efficacy of ethanol in adolescence, effects of stressors on this efficacy, and potential long-term consequences of adolescent exposure for later intake of ethanol. These issues will be examined using a model of adolescence in rats in which, during a compressed time period of around 2 weeks, developing rats exhibit certain characteristic neurobehavioral features of adolescence similar to those reported in adolescents of other species, including humans. Both males and females will be examined, due to reports of gender differences in ethanol responsiveness but the limited attention previously given this issue in animal studies with alcohol. Three specific aims are proposed: 1) to examine whether the reinforcing efficacy of ethanol is altered in adolescents relative to adults, and the role of chronic stress and corticosterone in contributing to age-related differences in ethanol reinforcement; 2) to establish whether alcohol intake in adolescence is more affected by stressors than such intake in adulthood; and 3) to determine whether prior exposure to ethanol and/or stress has a greater impact on subsequent ethanol consumption when that exposure occurs during adolescence than in adulthood. The results of this basic research should provide an essential initial base of data regarding ethanol reinforcement and stress during adolescence as well as potential long-lasting consequences of such exposure.

DESCRIPTION (provided by applicant): Human adolescents and their counterparts in a variety of animal models avidly seek out rewarding stimuli through heightened peer interactions, risk taking, and increases in food consumption and drug use. These behavioral features of adolescence are highly conserved across species, as are adolescence-associated transformations in forebrain regions (e.g., PFC, amygdala, accumbens, and associated DA input) implicated in these behaviors and in attributing hedonic value and incentive motivation to natural rewards, drugs of abuse, and their associated cues. A crucial core question remains, constraining understanding of problem behaviors in adolescence and attempts to discern their neural substrates: Are adolescent-associated increases in behaviors directed towards natural rewards and drugs of abuse related to increases or decreases in the value they attribute to these rewarding stimuli? On the one hand, adolescents might pursue certain natural rewards and drugs because they normally exhibit (or are prone to develop) strong incentive motivation for these stimuli. Alternatively, adolescents may avidly seek these rewards because they are attempting to compensate for an age-related insensitivity in reward circuits that produces a partial, developmentally expressed anhedonia. Using an established model of adolescence in the rat and focusing on three natural rewards of particular significance for adolescents: social interactions, novelty, and appetitive taste) stimuli, the proposed work will test these possibilities and determine age-specific expression of neural markers for these rewards. Work in this proposal will answer the following questions: Do adolescents exhibit attenuated hedonic affect to natural rewards relative to mature animals (Sp. Aim 1)? Do they express increased incentive motivation for natural rewards (Sp. Aim 2) or are they unusually prone to develop incentive sensitization to these rewards following chronic drugs or stressors (Sp. Aim 3)? Do adolescents exhibit unique patterns of regional brain activation in PFC, amygdala, accumbens and related circuitry in response to natural rewards and their cues when compared with mature animals (Sp. Aim 4)? The proposed work will further understanding of why adolescents behave the way they do, identify candidate neural regions underlying these age-related proclivities, and help inform strategies for the treatment of adolescents exhibiting drug abuse problems or other excessive reward-directed behaviors.

DESCRIPTION (provided by applicant): Women have been excluded or underrepresented in much past research examining alcohol use and dependence. Yet, the research available has found women to differ from men in their alcohol drinking patterns and their sensitivity to various consequences of ethanol. These sex differences begin to emerge during adolescence and become more pronounced at maturity. While social, cultural and environmental influences undoubtedly contribute to the emergence of sex differences in alcohol intake and effects, biological factors also are likely to play an important role. Yet, despite the marked hormonal and neural changes associated with puberty and the other physiological transitions of adolescence, little is known of how these factors might contribute to the ontogeny of sex differences in ethanol intake and sensitivity. Determining the role of pubertal increases in gonadal hormones on the emergence of sex differences in alcohol consumption and consequences is ethically challenging in humans, but can be approached using animal models that share with human adolescents certain basic age-typical similarities in hormonal, neural, and behavioral characteristics. The proposed work will use a simple animal model of adolescence in the rat to examine the role of pubertal increases in gonadal hormones in both males and females on the emergence of sex differences in ethanol intake and consequences. Given mounting evidence that pubertal rises in gonadal steroids can exert organizational effects on the brain to support the differentiation of certain sex-typical behaviors at puberty, both organizational as well as activational (direct hormone-stimulated) effects of gonadal hormones will be explored. This will be accomplished via gonadectomizing animals either pre-pubertally or in adulthood, and hormone replacement beginning either at puberty or maturity. Focusing on alcohol-related sex differences that begin to emerge during the adolescent period and become marked in adulthood, the proposed studies will assess both organizational and activational effects of the rise in gonadal hormones at puberty on expression of sex differences in ethanol intake, ethanol sensitivity (indexed by ethanol-induced social suppression) and ethanol stress interactions. Despite the increasing emphasis on alcoholism as a developmental disorder, little is known of the potential role of the rise in pubertal hormones in promoting the emergence of sex-typical patterns of alcohol use, consequences and abuse. The proposed work will address this considerable gap in our understanding, data that should help inform the development of sex-relevant strategies for prevention, identification, and treatment of alcohol use disorders among youth and young adults.

[unreadable] DESCRIPTION (provided by applicant): Adolescence is a time of notable neurobehavioral transitions, with increases in peer-directed social interactions common among adolescents of a variety of species. It is during adolescence that alcohol intake becomes normative in humans, with a significant number of adolescents engaging in high (binge) levels of use. Most adolescent use of alcohol occurs in a social context, with adolescents reporting that a key motive for their drinking is for social facilitation. Using an animal model of adolescence, adolescent rats likewise often drink substantially more than mature rats, and are uniquely sensitive to ethanol-induced increases in peer-directed social interactions. This ethanol-induced social facilitation is not normally evident in adult rats, but emerges in adults and becomes even more pronounced in adolescents following chronic stress. Despite the critical role that the social context plays in adolescent alcohol use, little is known of the social neuroscience of this use - e.g., whether ethanol enhances the rewarding properties of social stimuli to make social interactions particularly rewarding for adolescents, or whether ethanol exposure in a social context increases ethanol's hedonic impact among adolescents. These possibilities may not be mutually exclusive and will be explored, along with potential neural substrates for these effects, using an animal model of adolescence in the rat in the proposed work. Effects of ethanol on the rewarding value of social stimuli will be examined in Aim 1 through studies of social motivation (approach and anticipation), social affect, and social buffering, while Aim 3 will use similar measures to assess whether sensitization to ethanol-induced social facilitation induced by chronic stress is related to an enhancement of the effect of ethanol on social reward. Effects of a social context on ethanol's aversive and appetitively rewarding properties will be examined in Aim 2 through studies of place conditioning, along with assessing the impact of a social context on ethanol intake in a voluntary, limited access situation. Candidate brain regions potentially contributing to enhanced ethanol/social reward expression during adolescence will be identified in Aim 4. The proposed work will further understanding of how the social context influences adolescent alcohol use, identify candidate neural regions contributing to these adolescent-characteristic interrelationships, and help inform strategies for the use of social context as a protective, rather than a risk factor for adolescent alcohol use. [unreadable] [unreadable] [unreadable]

DESCRIPTION (provided by applicant): Development of tolerance to alcohol is thought to reflect a critical sign of problematic alcohol involvement, and is included as one of the diagnostic criteria for alcohol dependence in the DSM IV. Yet little is known of the relationship between tolerance development and levels of alcohol consumption, although it has been speculated that emergence of tolerance to aversive effects of ethanol may permit greater consumption, increasing alcohol exposure levels and concomitant problems. Exploring the relationship between ethanol tolerance and consumption is particularly critical during adolescence, with 12-21 yr. olds not only drinking substantially more per drinking occasion than adults, but also reporting a higher incidence of ethanol tolerance than any other age group. The studies outlined in this proposal will explore this relationship using a simple and well characterized animal model of adolescence in the rat, a model that is typified by 2-3 fold high levels of ethanol consumption, expression of markedly more acute tolerance (and under some instances, chronic tolerance), and an attenuated sensitivity to various aversive effects of alcohol when animals in this developmental phase are compared with adults. The proposed work will explore the development of acute and chronic tolerance to various aversive effects of ethanol and the relationships of these adaptations to ethanol consumption during the adolescent period and following comparable manipulations in adulthood. Neural systems influencing these adaptations and whether these adaptations are causal for elevated ethanol consumption during adolescence will also be explored. Work in this proposal will answer the following questions: What are the relationships among the emergence of acute, rapid and chronic tolerance for various aversive effects of ethanol in adolescents and adults (Sp. Aim 1)? Do chronic exposure regimens sufficient to induce chronic tolerance to these ethanol effects lead to increases in voluntary ethanol consumption in home cage and limited access tests, with this increased consumption being particularly marked in adolescence (Sp. Aim 2)? What role do NMDA and opioid receptor subsystems play in the emergence of acute and chronic tolerance to ethanol during adolescence (Sp. Aim 3)? And, finally, are the adaptive changes associated with chronic tolerance development causal for increasing ethanol consumption in adolescence (Sp. Aim 4)? The proposed work will not only provide convincing evidence as to whether the emergence of chronic tolerance to ethanol leads to increases in its consumption, but will also illuminate the relationships among acute, rapid and chronic tolerance and the role of tolerance development in contributing to the elevated ethanol consumption and the insensitivity to various aversive effects of ethanol characteristic of adolescence. PUBLIC HEALTH RELEVANCE: Development of alcohol tolerance is viewed as a sign of problematic alcohol involvement and is one of the criteria used to diagnose alcohol dependence, yet little is know of the relationship between tolerance development and levels of alcohol consumption. Exploring tolerance/consumption relationships is particularly critical during adolescence, given that 12-21 yr. olds not only drink substantially more per drinking occasion than adults, but they also report a higher incidence of alcohol tolerance than any other age group. The studies outlined in this proposal will use a simple and well-characterized animal model of adolescence in the rat to compare different forms of tolerance and how they change developmentally, explore the neural substrates of these adaptations to alcohol, and determine whether tolerance development is causal for inducing the high levels of alcohol consumption characteristic of the adolescent period.

DESCRIPTION (provided by the applicant): This P30 proposal is in response to RFA-OD-09-005 and has the goal of hiring an early stage investigator with a strong record of research productivity and interests and expertise in alcohol research of a developmental nature as a tenure-track, assistant professor in the Department of Psychology at Binghamton University. This new P30-assisted hire would (a) enhance and complement the emphasis on developmental and alcohol research within the behavioral neuroscience area of the department, as well as (b) broaden and strengthen the interdisciplinary research program on fetal/adolescent alcohol exposure that is the focus of the Developmental Effects of Alcohol Research Center (DEARC), a newly NIAAA-funded alcohol research center that unites researchers at SUNY Upstate Medical University, Binghamton University and other institutions in upstate New York. The new hire will not previously have held a tenure-track position, and will spend 75% of his/her time on research activities during the academic year, and 100% of time devoted to research during the summer months over the two years of this award. In addition to this salary support, support for start-up funds and for a graduate research assistant is also requested. During the two year grant period, the new faculty member would be expected to establish an independent research laboratory at Binghamton, submit research from his/her laboratory for peer-reviewed publication, initiate the research training of one or more graduate students, collect preliminary data for one or more NIH grant proposals on which he/she is PI, and obtain DEARC funding for a pilot project. The new hire would be assisted in meeting these expectations by multiple mentoring mechanisms within a program recognized for being highly supportive of junior faculty and a research setting characterized by excellent laboratory space, outstanding equipment resources, and abundant opportunities for collaboration provided by numerous productive colleagues working in similar areas.

Alcohol use by human adolescents is pervasive, with over 10% of 8th graders, 20% of 10th graders and 25% of 12th graders reporting consumption of five or more drinks in a row in the past two weeks. High levels of alcohol consumption may be fostered in adolescents in part because of their insensitivity to certain desired ethanol effects as well as to effects of ethanol used to self-monitor intake, whereas these relatively high intakes may exacerbate certain adverse ethanol effects to which adolescents may be unusually sensitive. Indeed, animal studies have shown adolescents to differ considerably from adults in their sensitivity to acute ethanol, with these alterations bi-directional in nature. Though less sensitive than adults to many ethanol effects (e.g., sedative, motor-impairing and anxiolytic effects), adolescents are conversely more sensitive than their adult counterparts to ethanol-induced social facilitation and ethanol-related impairments in LTP and spatial memory. Recent work has shown that these effects are particularly pronounced during early adolescence, with animals of this age varying considerably in their ethanol sensitivities from older adolescents and adults. Using an established animal model of adolescence and animals generated from the ANIMAL CORE, the proposed work will examine factors contributing to the unique mosaic of behavioral sensitivities to ethanol seen in young adolescents. Psychopharmacological studies will explore contributions of developmental alterations in GABAA and NMDA receptor systems to an ethanol effect for which young adolescents are unusually sensitive (social facilitation) as well as for consequences of ethanol to which they are relatively resistance (anxiolysis and behavioral suppression) when compared with older adolescents and adults. Using the NEUROANATOMY CORE, additional studies will examine ontogenetic patterns of regional brain activation undergoing transformation during adolescence, and determine whether pharmacological manipulations that block age-related behavioral responses to ethanol likewise attenuate age-specific patterns of ethanol-induced cFos activation in target regions of interest.

DESCRIPTION (provided by applicant): Adolescence is a time of considerable transformation in stress-sensitive brain regions that play key roles in the processing of rewarding and social/emotional stimuli and in areas that exert regulatory control over these regions. Given that lasting effects of substances are most likely to occur when systems sensitive to those substances are changing developmentally, we hypothesize that adolescent intermittent ethanol (AIE) exposure will induce lasting effects on social behavior and on the social rewarding and aversive effects of ethanol, leading to persistent elevations in adult intake due in part to alcohol's social anxiolytic properties. Indeed, social rewarding effects of alcohol are critical for the initiation of alcohol use in adolescence and escalation to heavy drinking among individuals with social anxiety, with socially anxious alcoholics showing greater levels of social impairment than non-dependent, socially anxious individuals. The proposed studies will investigate the consequences of voluntary and experimenter-administered adolescent intermittent alcohol (AIE) on later social anxiety and reward, the efficacy of alcohol for countering these lasting perturbations in social behavior, and the impact on later voluntary intake of ethanol, along with neural/genetic adaptations underlying these effects. Using synergisms provided by the NADIA, the proposed research will address the following aims: (1) test whether AIE increases baseline levels of anxiety-like behavior under social circumstances in adulthood and exacerbates stress-induced social anxiety, and determine the efficacy of acute ethanol for reversing this social anxiety;(2) determine whether AIE exposure disrupts the rewarding and/or hedonic value of social stimuli, whether these alterations are further exacerbated by stress, and the efficacy of acute ethanol for reversing these effects;(3) assess whether AIE enhances ethanol drinking in adulthood and decreases sensitivity to the aversive properties of ethanol, and whether these effects of AIE are further exacerbated by social context in adulthood;(4) investigate AIE effects on dopamine transmission during adulthood by measuring real-time dopamine release in nucleus accumbens in response to social stimuli and assessing dopamine-related gene expression in key limbic regions. PUBLIC HEALTH RELEVANCE: The proposed investigations of lasting effects of adolescent alcohol exposure on social anxiety, alcohol's anxiolytic properties, and underiying neural/genetic adaptations are of considerable relevance, given the importance of alcohol's social rewarding properties in initiating use and escalating to heavy drinking among individuals with social anxiety, along with the greater social impairment seen in socially anxious alcoholics.

DESCRIPTION (provided by applicant): The Administrative Core provides leadership and support to promote the highest quality research in the DEARC and its impact. The goals of the Administrative Core are to facilitate interactions among projects and attain research excellence throughout the DEARC by providing clear leadership, efficient management, effective committee structure, strong administrative, fiscal and research infrastructure, and the promotion of integration and synergism. These goals are supported through 5 aims. Aim 1 provides an organizational infrastructure to facilitate attainment of Center goals and objectives through a strong Executive Committee (EC) consisting of the PI (L. Spear) and the two co-l's (T. Deak & S. Youngentob) of the Administrative Core, aided by an efficient DEARC office and administrator, capable fiscal support through the State University of New York Research Foundation (with offices at both Binghamton University [BU] and Upstate Medical University [UMU]), as well as strong institutional support and an impressive research infrastructure that includes new animal buildings on both DEARC campuses. Building on the culture of collaboration within the DEARC, Aim 2 promotes communication, facilitates interactions and encourages integration and synergisms across the DEARC through such activities as monthly meetings, mini-retreats, DEARC-sponsored colloquia, and a yearly retreat. Aims 3 and 4 are to ensure oversight, coordinate interactions among research projects and cores, promote research-supportive allocation of resources, and utilize internal and external guidance committees to ensure quality and progress of research components and cores. Day-to-day decisions will be made by the EC with frequent consultation of a Steering Committee (composed of PIs of cores and main projects) and oversight by two critical guidance committees: an Internal Advisory Board (lAB) comprised of administrators at BU and UMU, and an External Advisory Board (EAB) comprised of nationally prominent senior alcohol researchers who serve as a program advisory committee to evaluate and advise the DEARC regarding all aspects of Center functioning. Aim 5 is to encourage enrichment at all career levels, promote the recruitment of new researchers in the field of alcohol and development, and enhance outreach efforts.

Alcohol use by human adolescents is pervasive, with over 10% of 8th graders, 20% of 10th graders and 25% of 12th graders reporting consumption of five or more drinks in a row in the past two weeks. High levels of alcohol consumption may be fostered in adolescents in part because of their insensitivity to certain desired ethanol effects as well as to effects of ethanol used to self-monitor intake, whereas these relatively high intakes may exacerbate certain adverse ethanol effects to which adolescents may be unusually sensitive. Indeed, animal studies have shown adolescents to differ considerably from adults in their sensitivity to acute ethanol, with these alterations bi-directional in nature. Though less sensitive than adults to many ethanol effects (e.g., sedative, motor-impairing and anxiolytic effects), adolescents are conversely more sensitive than their adult counterparts to ethanol-induced social facilitation and ethanol-related impairments in LTP and spatial memory. Recent work has shown that these effects are particularly pronounced during early adolescence, with animals of this age varying considerably in their ethanol sensitivities from older adolescents and adults. Using an established animal model of adolescence and animals generated from the ANIMAL CORE, the proposed work will examine factors contributing to the unique mosaic of behavioral sensitivities to ethanol seen in young adolescents. Psychopharmacological studies will explore contributions of developmental alterations in GABAA and NMDA receptor systems to an ethanol effect for which young adolescents are unusually sensitive (social facilitation) as well as for consequences of ethanol to which they are relatively resistance (anxiolysis and behavioral suppression) when compared with older adolescents and adults. Using the NEUROANATOMY CORE, additional studies will examine ontogenetic patterns of regional brain activation undergoing transformation during adolescence, and determine whether pharmacological manipulations that block age-related behavioral responses to ethanol likewise attenuate age-specific patterns of ethanol-induced cFos activation in target regions of interest.

The success of the DEARC depends upon the ability of the ADMINISTRATIVE CORE to facilitate interactions and attain the highest quality research among the projects. These goals will be achieved through multiple mechanisms. (1) The quality, progress, and fiscal integrity of research components (i.e., MAIN and PILOT PROJECTS) will be assured through both internal and external guidance systems, (a) Two internal systems will be an Executive Committee (formed by the Center Director and Scientific Director) and a Coordinating Committee (comprised of the Pis of the Main Projects and the Cores). Pis will report to these committees through regular written reports and orally at monthly meetings and at the annual DEARC Retreat, (b) Two extra-DEARC bodies will oversee the activities of the DEARC: an Internal Advisory Board comprised of administrators at Binghamton Univ., SUNY- Cortland, Syracuse Univ., and Upstate Med. Univ. and an External Scientific Advisory Board comprised of nationally prominent, senior alcohol and developmental biology researchers. (2) A critical facet of the DEARC will be the three Scientific Cores (ANIMAL, CELL/MOLECULAR, and NEUROANATOMY CORES). The two internal systems described above will oversee the CORES, assuring the quality of the data generated by the CORES and the productivity of the CORES. (3) DEARC researchers are located at three SUNY campuses- in Binghamton, Cortland, and Syracuse. The ADMINISTRATIVE CORE will assure that handling of samples and data is efficient and that communication between these campuses is seamless. These will be achieved using wellestablished inter-campus courier services, electronic communications, and sophisticated teleconferencing systems. (4) The PILOT PROJECT PROGRAM is designed to generate new ideas and new programs. The ADMINISTRATIVE CORE will guide this program. This involves soliciting and reviewing applications and monitoring the progress of the funded projects. (5) The ADMINISTRATIVE CORE will support the continuing education of DEARC personnel. This will occur through journal clubs, team-taught courses, and seminar series. These offerings will be made available to more people through teleconferencing and videotaping (archiving). Other communications by DEARC personnel (e.g., poster, manuscript, and grant preparation) will be supported by the Administrative Core. Together, the ADMINISTRATIVE CORE will serve as a system for maintaining quality and progress of the DEARC and supporting the communication within and beyond the DEARC.

A primary goal of the DEARC is to elucidate the brain and behavioral consequences of ethanol exposure throughout a broad range of developmental stages encompassing gestation through adulthood. All projects described in this center proposal utilize rodent models, making the Animal/Behavior Core a critical component of the DEARC. The Animal/Behavior Core plays several key roles within the center: It provides guidance and training on breeding practices, ethanol exposure, surgical techniques and behavioral phenotyping. In addition, the Animal/Behavioral Core provides access to equipment for behavioral assessment and measurement of ethanol in blood and brain. Another major goal of the Animal/Behavior Core is to facilitate the sharing of animal resources through an interactive on-line database. These goals will be facilitated by the core PI and Col, each with more than a dozen years of experience. The Animal/Behavior Core will be critical in maintaining the over arching goal of the DEARC of cross-study comparisons to understand how developmental ethanol exposure changes brain and behavioral function across the lifespan. Specifically the Animal/Behavior Core will streamline the generation, use, and distribution of animal resources, while at the same time providing crucial support for the standardization of techniques, thereby assuring quality control. Finally, the Animal/Behavior Core will play an essential role in promoting cross talk among Pis and between the Pis and the Administrative Core.

High levels of ethanol (EtOH) consumption during adolescence may be in part biologically driven, given that adolescents often manifest 2-3 fold greater EtOH intakes than adults in a variety of mammalian species. Studies in rats have suggested that such elevated intakes may be possible in part because adolescents are less sensitive than adults to aversive effects of EtOH and other intoxicating effects that may normally serve as cues to moderate intake. Genetic studies have found that sensitivity to aversive effects of EtOH plays an important role in moderating EtOH intake, and is more strongly related to EtOH intake than sensitivity to EtOH's rewarding properties. Despite compelling evidence for an association between insensitivity to EtOH aversion and enhanced EtOH intake, little is known about the ontogeny of these aversive sensitivities, their regional specificity and neural contributors. In conjunction with the DEARC NeuroCore and Animal/Behavior Core, the proposed work will address these critical gaps. Aim 1 will use conditioned taste aversions (CTA) to examine the ontogeny of the developmental insensitivity to EtOH's aversive effects throughout the juvenile to adult period. Aim 2 will assess c-Fos activation patterns to aversive effects of EtOH across age and associate these activation patterns with age differences in EtOH aversion. Focusing on an aversive region where activation is strongly linked to age differences in EtOH CTA, Aim 3 will determine whether reversible inactivation of that region diminishes age differences in EtOH's aversive effects and EtOH intake differences, while Aim 4 will examine potential neural substrates underlying the age differences in EtOH-induced activation in this region via examination of NMDA, GABA and opiate receptor regulation and intracellular signaling pathways. Aim 5 will determine whether elevations in adolescent drinking induced by prenatal EtOH exposure are accompanied by exacerbated insensitivies to EtOH's aversive effects. Collectively, these studies will provide critical new information to further our understanding of how ontogenetic changes in the brain contribute to adolescent insensitivities to aversive properties of EtOH, and ultimately to the elevated levels of EtOH drinking characteristic of adolescence.

The Administrative Core provides leadership and support to promote the highest quality research in the DEARC and its impact. The goals of the Administrative Core are to facilitate interactions among projects and attain research excellence throughout the DEARC by providing clear leadership, efficient management, effective committee structure, strong administrative, fiscal and research infrastructure, and the promotion of integrafion and synergism. These goals are supported through 5 aims. Aim 1 provides an organizational infrastructure to facilitate attainment of Center goals and objectives through a strong Executive Committee (EC) consisting of the PI (L.Spear) and the two co-l's (T.Deak & S.Youngentob) of the Administrative Core, aided by an efficient DEARC office and administrator, capable fiscal support through the State University of New York Research Foundation (with offices at both Binghamton University [BU] and Upstate Medical University [UMU]), as well as strong institufional support and an impressive research infrastructure that includes new animal buildings on both DEARC campuses. Building on the culture of collaboration within the DEARC, Aim 2 promotes communication, facilitates interactions and encourages integration and synergisms across the DEARC through such activities as monthly meetings, mini-retreats, DEARC-sponsored colloquia, and a yearly retreat. Aims 3 and 4 are to ensure oversight, coordinate interactions among research projects and cores, pronriote research-supportive allocation of resources, and utilize internal and external guidance committees to ensure quality and progress of research components and cores. Day-to-day decisions will be made by the EC with frequent consultation of a Steering Committee (composed of Pis of cores and main projects) and oversight by two critical guidance committees: an Internal Advisory Board (lAB) comprised of administrators at BU and UMU, and an External Advisory Board (EAB) comprised of nationally prominent senior alcohol researchers who serve as a program advisory committee to evaluate and advise the DEARC regarding all aspects of Center functioning. Aim 5 is to encourage enrichment at all career levels, promote the recruitment of new researchers in the field of alcohol and development, and enhance outreach efforts.

? DESCRIPTION (provided by applicant): Our current work in the NIAAA consortium on the Neurobiology of Adolescent Drinking in Adulthood (NADIA) has yielded four key findings of critical importance regarding the consequences of adolescent intermittent ethanol (AIE). These consequences: a) differ dramatically with exposures during early-mid versus late adolescence; b) are sex-specific; and include c) induction of social anxiety, and d) the persistence of adolescent-typical phenotypes, including adolescent-typical responses to ethanol into adulthood. Our current proposal will address critical gaps arising from this work. Aim 1 will assess whether early-mid adolescent AIE exposure (analogous to early initiation of use in adolescence) alters social/affective/reward domains influenced by subcortical limbic systems whose activity is particularly prevalent at that time, whereas AIE in late adolescence (analogous to the late adolescent/emerging adulthood period in humans where binge drinking is particularly pronounced) produces alterations in tasks requiring integrity of late maturing prefrontal areas. Effects are predicted to be sex-specific, with early AIE effects more pronounced in males and late AIE consequences in females. Aim 2 will focus on neural substrates underlying the marked social anxiety-like behavior induced by early AIE in males, with a focus on two peptides critically involved in social behavior: oxytocin (OXT) and vasopressin (AVP). Psychopharmacological approaches along with assessment of levels of OXT, AVP and their receptors, as well as associated epigenetic modifications in two key regions involved in social anxiety (amygdala and hypothalamus), will be used to test the hypothesis that social anxiety in males is associated with a shift in balance of OXT and AVP toward AVP. Lastly, Aim 3 will assess whether the persistence of well-characterized adolescent-typical ethanol sensitivities into adulthood following AIE relates to perturbations in excitatory-inhibitory balance. Both psychopharmacological and molecular approaches will be used to test this hypothesis, with a particular emphasis on glutamate NR2B and AMPA and extrasynaptic GABAA receptors along with vesicular transporter ratios. Studies in both Aims 2 and 3 will be conducted with an eye toward identification of pharmacotherapeutic approaches to reverse AIE effects.

ABSTRACT The goal of this proposal is to support a new pre- and post-doctoral training program in the area of development and neuroadaptations in alcohol and addictions (DNA2). Given increasing recognition that alcoholism and other substance use disorders have roots established in development, the proposed training grant will provide the next generation of addiction researchers with crucial training in the conceptual issues and methodological approaches for studying developmental neuroadaptations leading to abuse and addiction. Building on existing research strengths in this area exemplified by the Developmental Exposure Alcohol Research Center (DEARC) and anchored by historically strong graduate training programs in development, addictions and neuroadaptations within psychological sciences along with rapidly evolving programs in community science and public affairs, the DNA2 T32 provides the rich, cooperative intellectual environment conducive for highly successful research training, didactic education and mentorship for the proposed predoctoral (4/yr) and postdoctoral (2/yr) trainees. The training program emphasizes five key goals: (1) to actively seek and recruit a highly qualified and diverse group of pre- and post-doctoral trainees whose interest in and commitment to research in alcohol and addictions fits the laboratory(s) of one or more mentor-eligible faculty; (2) to provide individualized training experiences that foster expertise in state-of-the-art assessments, hypothesis formation, rigorous experimental design, data acquisition and analysis, and oral presentation and publication of results; (3) to assist trainees in generating preliminary data, novel hypotheses and specific aims and provide training in writing and grantsmanship for individual grant or F31/32 fellowship applications; (4) to create an individualized, multi-level research training environment that encourages both expertise and breadth in conceptual knowledge and technical capabilities and fosters translational considerations; and (5) to support the career/professional development of each trainee. To meet these goals, the T32 program will provide comprehensive interdisciplinary training through didactic coursework, trainee workshops, seminars/colloquia, journal and writing clubs, supervised research experiences, and participation in professional meetings and extra-university training opportunities. Methodological issues and techniques in alcohol and drug abuse research will be emphasized throughout the typical 2-year training period for each trainee, with a particular focus on training in developmental approaches, and rigorous attention to education in the responsible conduct of research, including means to enhance research reproducibility. This training program combines the expertise of faculty with established expertise in the alcohol field as well as in research with other drugs of abuse to capitalize on synergies across research programs and perspectives.

PROJECT SUMMARY/ABSTRACT MAIN RESEARCH COMPONENT 4 Alcohol is one of the most widely used substances by American adolescents, with binge and heavy drinking evident in almost two thirds of underage current drinkers. These high rates of binge and heavy drinking are alarming, since adolescents who engage in even episodic heavy drinking are more likely to exhibit alcohol use disorders and other adverse consequences later in life. Young people predominantly drink in social situations, although this context specificity ? let alone sex differences in sensitivity to social consequences ? has been little investigated. Using a rat model of adolescence, we have shown pronounced qualitative sex differences in the precursors and effects of ethanol contributing to high social drinking among adolescents. High social drinking among males is associated with high social activity and enhanced sensitivity to the socially facilitating effects of ethanol, whereas in adolescent females, higher levels of social drinking are associated with elevated social anxiety and enhanced sensitivity to the socially anxiolytic effects of ethanol. The present proposal is designed to separately determine brain regions that are responsible for high social activity and sensitivity to ethanol-induced social facilitation in adolescent males, in contrast to regions related to high social anxiety and enhanced sensitivity to the socially anxiolytic effects of ethanol in females. Target brain regions that are differentially activated in males and females with high social drinking phenotypes will be determined using transgenic cFos-LacZ rats and X-Gal staining for c-Fos. The Daun02 procedure will then be used to selectively inactivate neuronal ensembles in specified target regions that were activated by the social stimulus alone or in combination with acute EtOH, and consequences of this inactivation on subsequent social drinking determined in male and female high social drinkers. We expect that inactivation of neural ensembles activated by social interactions alone or in combination with acute EtOH in high socially active males will attenuate social drinking in these animals, whereas inactivation of neuronal ensembles activated in high socially anxious adolescent females by social stimuli will diminish social drinking in these females. Given the critical importance of brain vasopressin/oxytocin peptide systems in regulation of social behavior and social anxiety, we will also test the hypothesis that high social drinking in males is associated with hyperactivity of the brain vasopressin V1a receptor, whereas functional hypoactivity of the brain oxytocin system contributes to high social drinking in adolescent females. These hypotheses will be tested neuropharmacologically and via assessment of protein levels for oxytocin and vasopressin and their receptors in the brain regions sex-specifically associated with high drinking phenotypes. The work outlined in this proposal will be among the first to examine neural contributors to the pronounced qualitative sex differences in precursors leading to high social drinking among adolescents, and are essential for creation of new, sex-specific early prevention and intervention strategies for heavy alcohol use in adolescence.