Terrence Deak - US grants

In recent years, there has been increasing recognition that pro- inflammatory cytokines play a role in the behavioral and physiological alterations produced by exposure to psychological stressors. The data to support this conclusion comes from the findings that (a) acute stress can induce pro-inflammatory cytokine production both centrally and peripherally, (b) administration of pro-inflammatory cytokines produces many of the same behavioral and physiological consequences as acute stress exposure, and (c) blockade of pro-inflammatory cytokine activity inhibits or attenuates some behavioral and physiological consequences of acute stressor exposure. These data have led to the suggestion that production of cytokines in response to stress may provide novel insight into the etiology of stress-related disorders. Perhaps one of the most widely used animal models of stress is the forced swim test. During this stressor, rats are forced to swim in a cylinder of water from which there is no escape. After an initial period of behavioral activation characterized by repeated attempts to escape, rats then adopt a characteristic posture of immobility that has been conceptualized as behavioral despair. Given recent notions that cytokines mediate some stress-related behaviors, we explored whether the pro-inflammatory cytokine Interleukin01 (IL-1) would be elevated in rats that have been forced to swim. Our preliminary results demonstrate a robust increase in IL-1 in all peripheral tissues examined (blood, pituitary, and spleen) immediately after the forced swim session. These data raise the possibility that the immobility response observed during the forced swim stressor may be mediated by IL-1. Thus, the goal of this proposal is to characterize the time-course of the swim stress induced increase in IL-1 and determine whether the increase in IL-1 is functionally related to the changes in behavior observed during forced swim exposure.

While neurons were once considered to be the primary functional cells of the nervous system, it has become increasingly clear in recent years that other cell types such as astrocytes and microglia powerfully modulate the activity of neurons. For instance, microglia have always been viewed as passive cells in the brain that modulate neuronal activity only when challenged by an inflammatory stimulus or traumatic brain injury. This project challenges the existing notion that microglia reside in a quiescent state in the normal brain, suggesting that the principal biological response to stress is under tight regulatory control by microglia. This project implicates microglia as a novel cellular target for regulation of the neuroendocrine stress response, providing a role for microglia in negative feedback mechanism. The project examines the role of specific brain structures where microglia are thought to exert inhibitory control over the neuroendocrine stress response and examines one potential cellular mechanism by which microglia may regulate thse neuroendocrine stress responses. The project uses a combination of biochemical and anatomical procedures to address these issues. Together, these projects will forge an altogether new understanding of how diverse cell groups interact within the central nervous system and is expected to produce a paradigm-shift in how investigators view basic regulation of the stress response. This project provides a unique opportunity for training of undergraduate and graduate students from a diverse range of backgrounds (50% women, 33% minority). The laboratory environment is state-of-the-art and contains extensive resources devoted specifically to promote training and networking for careers in science.

[unreadable] DESCRIPTION (provided by applicant): Withdrawal from acute alcohol exposure produces profound changes in behavior including reduced exploration and locomotor activity as well as suppressed social interaction. These behavioral changes characterize the "hangover" state and are remarkably similar to sickness behaviors that are typically observed during acute illness produced by infectious agents or injection of pro-inflammatory cytokines. We therefore propose that behavioral responses during withdrawal from acute alcohol exposure in rats may be viewed as sickness-like behavioral responses that are initiated and maintained by activation of inflammatory-related pathways traditionally associated with immune activation. Our preliminary data indicate robust increases in transcriptional activity of interleukin-1 in CNS sites that are crucial for normal progression of the immune response and the generation of sickness behaviors. The goal of the following proposal, therefore, is to (Aim 1) establish the temporal and spatial characteristics of cytokine responses produced by hangover, (Aim 2) to identify probable cellular and neurochemical mechanisms of cytokine responses produced by hangover, and (Aim 3) to forge a positive mechanistic link between increased central cytokines and the sickness-like behavioral changes observed during hangover in rats. [unreadable] [unreadable] [unreadable]

Exposure to stress leads to activation of multiple systems such as the sympathetic nervous system (SNS) and the hypothalamic-pituitary-adrenal (HPA) axis, both of which evolved as endogenous mechanisms to promote survival in the face of diverse threats. However, considerably less attention has been devoted towards understanding brain mechanisms that promote functional recovery and adaptation after the threat subsides. As a result, we have argued that exposure to intense stressors requires a protracted period of recuperation, and that activation of inflammatory pathways in the brain as a result of stressor exposure promotes recuperative behavior. The goal of this project, therefore, is to determine the mechanisms by which stressor exposure leads to activation of inflammatory pathways in the brain. The researcher specifically hypothesizes that activation of central noradrenergic nuclei in response to stress are critical for activating these inflammatory pathways, while the release of corticosterone, the major effector molecule of the HPA axis in rat, constrains the development of neuroinflammation. In this regard, the project employs a systems biology approach that seeks to clarify the functional interplay among multiple stress responsive systems at the cellular, neurochemical, and neuroanatomical levels of analysis. The investigator's laboratory is comprised of trainees at all levels of the scientific hierarchy (undergraduates, graduate students and post doctoral trainees) and includes strong representation by underrepresented populations in the sciences (50% female, 33% minorities). The laboratory environment is state-of-the-art and contains extensive resources devoted specifically to promote training and networking for careers in science. The laboratory also has a strong history, and deliberate plans for, dissemination of findings to the scientific community (both domestic and abroad) as well as to the lay public.

A primary goal of the DEARC is to determine the consequences of exposure to ethanol during development on brain structure and function. Development is defined as encompassing the gestational period through adolescence, inclusive. All projects described in this application will use rodents (Sprague- Dawley rats and mice) as a model;thus, the ANIMAL CORE is a critical component for the DEARC. The ANIMAL CORE has multiple roles within the Center. It provides a rodent breeding program, standardized modes of ethanol administration (and administration of other drugs), standardized maintenance of pregnant, pre-weanling, and adolescent rodents, standardized determination of blood and brain ethanol concentrations (BECs and BrECs, respectively), collaboration in the design of experimental paradigms, and maintenance of a database containing maternal/litter data. As all animal research will be performed in Binghamton (Binghamton University) and Syracuse (Upstate Medical University), there are two components to the ANIMAL CORE. Each subdivision will be supervised by a member with more than a dozen years of experience with animal models, many of those years performing alcohol research. The subdivisions offer complementary, non-overlapping models of ethanol exposure. Having a coordinated ANIMAL CORE increases the efficient use of animals and standardizes the generation of animals. This is critical for the generation of compatible models that are the basis for the proposed anatomical, biochemical, molecular biological, and behavioral studies. To this end, the ANIMAL CORE will purchase, house, and breed animals. In addition, BECs and BrECs will be determined and databases following the subjects (cradle-to-grave) will be maintained by the ANIMAL CORE.

Activation of inflammatory signaling pathways within the CNS, including increased expression of proinflammatory cytokines, microglial activation, and other tell-tale signs of neuroinflammation, have emerged as powerful drivers of ethanol-dependent behavioral change and neuropathological consequences of ethanol exposure. While ethanol load is certainly a critical determinant of ethanol-induced cytokine changes, virtually nothing is known about how ethanol-induced inflammatory processes are impacted by cues associated with ethanol exposure, or how ethanol-induced inflammatory processes differ across key stages of development, particularly the juvenile-* adolescent?> adult continuum, where age of ethanol initiation is most predictive of later Alcohol Use Disorders (AUD). As such, the overarching goal is to identify mechanisms of plasticity in central cytokine responses evoked by ethanol challenge across eariy development in a rodent model. The proposed work will significantly advance our understanding of ethanol-induced (2-4 g/kg) expression bf Interieukin-6 (IL-6), a rapidly induced (3 hr post ethanol) cytokine effect that occurs in key CNS structures relating to learning and memory (hippocampus) and aversive processes (paraventricular nucleus; PVN). Importantly, when ethanol was paired with novel cues (lemon odor in a unique context) across 4 trials, a substantially lower dose of ethanol (0.5-g/kg) on the fifth trial led to a robust increase in IL-6 expression in the hippocampus (but not PVN). This context-dependent sensitization of IL-6 suggests that neuroinfiammatory consequences of ethanol exposure can come under the conditioned control of cues associated with ethanol exposure, thereby providing novel insight into how ethanol-related inflammatory processes transform across the first few ethanol exposures. Three Specific Aims will (1) characterize developmental differences in ethanol-dependent changes in IL-6; (2) identify developmentally critical/sensitive periods for ethanol-induced plasticity; and (3) determine cellular mechanisms of ethanol dependent expression of IL-6. The proposed work will significantly advance our understanding of how ethanol-related inflammatory processes transform as a function of early developmental exposure to alcohol.

DESCRIPTION (provided by applicant): Aging is a multifactorial process, in which a variety of neurobehavioral functions including social motivation and social recognition abilities decline at different rates, yet very little is known about the neural basis of these deleterious changes. The overarching goal of this proposal is to evaluate the neural mechanism(s) underlying the disintegration of social behavior during the natural course of aging, using rats as a model system. We hypothesize that the gradual decline in social behavior during senescence is symptomatic of both reduced motivational drive toward, and impaired recognition of, social stimuli. We argue that two established consequences of aging - a progressive decline in circulating androgen levels, termed andropause, and a gradual transition of the CNS toward a heightened state of pro-inflammation - play a key role in the disintegration of social behavior during senescence. The proposed mechanism is that gradual loss of circulating androgens leads to disinhibition of pro-inflammatory cytokines such as interleukin-1, which short-circuits th release of oxytocin (OT) expression in the paraventricular nucleus (PVN) of the hypothalamus and release in the medial amygdala (MeA), a key requisite for social motivation/recognition processes. Thus, Specific Aim 1 will characterize the age-related nature of social behavior deficits using a series of social behavior tasks, while at the same time determining the role of OT neurons in the MeA on the degradation of social behavior in aged rats. Specific Aim 2 will test the impact of the CNS pro-inflammatory state on the blunted response of social behavior circuitry and the therapeutic effect of administration of anti-inflammatory agents in aged rats. Specific Aim 3 will determine the role of gonadal hormones on the transition of the CNS toward a pro-inflammatory state and its impact on social behavior and their respective mechanisms of action. The outcome of the proposed work will integrate several core features of the aging process (declining androgens, heightened inflammation, altered neuropeptide regulation) into a meaningful mechanistic portrait of how social behavior erodes across the lifespan, and offer several specific therapeutic targets for reversing this process. As a result, the present studies hold great promise for translating findings from rodent models into direct improvements in the quality of life for aging populations.

The overarching goals of the Pilot Project Core are to (i) provide a structured funding mechanism to seed the development of great new ideas relevant to central themes of the DEARC; (ii) support career growth for both young and seasoned investigators through mentoring and facilitated collaboration; and (iii) capitalize on unique and timely opportunities that may arise from the rapidly changing, dynamic field of research on alcohol and development. The Pilot Project Core will achieve these goals through administration of a Pilot Project program that solicits Pilot Project Proposals from eligible investigators, coordinates the review process for proposals received, and ultimately oversees the award and reporting functions for all Pilot Projects. In addition, the Pilot Project Core, in close coordination with the Executive Committee, will help promote scientific connections among new members of the DEARC, and between these new members and established DEARC investigators to further increase scientific synergy and promote both peer-based and mentor-mentee relationships within the Center. Pilot Projects will be for 2 years each and funded at $30k (direct costs) each year. Using this approach, we will have a total of 6 funded pilot projects during the proposed 5-year funding period. Two highly innovative Pilot Project Proposals are proposed to begin in FY1. Pilot Project 1 (Cameron, PI) will examine the impact of maternal care during the neonatal period on DNA methylation and GABA-A receptor subunit expression, and how maternal care influences ethanol sensitivity later in life. Pilot Project 2 (Doremus-Fitzwater, PI) will examine the effect of adolescent ethanol exposure on coping behavior during aversive test situations, hypothesizing that adolescent ethanol exposure may differentially impact active versus passive coping strategies relative to adults. Through support of exciting new projects and promising investigators, the Pilot Project Core will serve as an incubator of great new ideas related to the central themes and mission of the DEARC.

The Developmental Exposure Alcohol Research Center (DEARC) is a successful, rapidly evolving alcohol research center that has been funded by NIAAA since 2009. The DEARC is an integrative, multidisciplinary center located at Binghamton University, one of four University Centers within the State University of New York (SUNY) system. The DEARC is unique in its focus on basic research to examine functional effects and neural consequences of ethanol exposure throughout extended periods of central nervous system development - inquiries that are of particular importance given the increasing recognition that alcoholism is a disorder with strong roots in development. Through the use of integrative research teams with broad interdisciplinary expertise and experience, the DEARC is distinctively positioned to focus its research efforts on determining the interactions between neurodevelopmental processes and alcohol, with a particular focus on two key points in development when the immature nervous system is particularly likely to be exposed to alcohol: prenatally through maternal alcohol consumption and during adolescence, when many youth first initiate alcohol use. The DEARC will include four Main research components that use established rodent models and integrative cellular, molecular, genomic, neuroanatomical, physiological and behavioral studies to explore functional and neural consequences of developmental exposures to alcohol. Projects also include an emphasis on sex-specific neurobehavioral processes, such as social context and anxiety, that promote alcohol consumption during adolescence. Pilot projects will be vetted and supported by a Pilot Project Core designed to further strengthen the DEARC while encouraging the development of new alcohol researchers. Critical support for all projects is provided by thea Behavioral Resources, Analysis and Integative Neuroscience (BRAIN) Core to support and integrate projects with genomic, molecular, and neuroanatomical expertise. Management, oversight and coordination of scientific effort is provided by an Administrative Core. The over-arching theme for these projects is that exposure of the developing nervous system to alcohol alters neural development and influences later responsiveness to alcohol in ways that exacerbate the normal propensity for enhanced alcohol use during adolescence, thereby increasing the probability of alcohol use and abuse in adulthood.

PROJECT SUMMARY/ABSTRACT ADMINISTRATIVE CORE The Administrative Core provides leadership and support to promote the highest quality research in the Developmental Exposure Alcohol Research Center (DEARC) and its scientific 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 to promote integration of projects and synergistic outcomes. 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, aided by an efficient DEARC office and administrator, capable fiscal support through the State University of New York Research Foundation, as well as strong institutional support and an impressive research infrastructure. 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 center leadership with frequent consultation of a Steering Committee and oversight by two critical guidance systems: an Internal Advisory Board (IAB) comprised of senior administrators at Binghamton University, and a Program Advisory Committee (PAC) comprised of five prominent senior alcohol researchers who serve as an external scientific advisory board 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 by DEARC investigators. The Administrative Core is therefore critical for seamless and effective integration of projects, as well as smooth and efficient operations of the DEARC as a whole.

PROJECT SUMMARY/ABSTRACT MAIN RESEARCH COMPONENT 3 Alcohol use and abuse represents a substantial threat to public health. Age of first alcohol exposure is a critical determinant of developmental trajectory and subsequent health status later in life, with prenatal and adolescent periods emerging as developmental epochs during which alcohol exposure is particularly prevalent. Neuroimmune consequences of alcohol have emerged as novel mechanisms that may contribute to changes in alcohol reinforcement, dependence, and ultimately the development of alcohol-related brain damage. Importantly, exposure to acute, binge-like doses of ethanol (EtOH) in rodents produce time-dependent changes in cytokine expression in which Interleukin-6 (IL-6) is substantially elevated in key limbic structures (amygdala, PVN and hippocampus) during acute EtOH intoxication. In contrast, expression of both IL-1? and TNF? tends to surge in these same structures during withdrawal from acute EtOH exposure. Surprisingly, adolescent rats (P31-33) displayed severely reduced cytokine responses to acute EtOH intoxication. These findings suggest that adolescent rats may have a functionally immature neuroimmune response relative to young adults. Paradoxically, however, adolescent Chronic Intermittent EtOH (CIE) exposure sensitized the intoxication-related IL-6 response evoked by a binge-like dose of EtOH later in life (P70 young adults). Thus, adolescents appear to be less sensitive to acute EtOH-induced cytokine responses, while at the same time being vulnerable to long-term sensitization of neuroimmune processes resulting from adolescent CIE exposure. However, the functional significance of these acute, EtOH-induced cytokine changes observed across the intoxication-withdrawal cycle remain obscure. This proposal will utilize contextual fear conditioning procedures as an animal model of emotional learning, and to test the functional relevance of EtOH-dependent expression of cytokines. Consistent findings demonstrate that EtOH impairs fear conditioning when training occurs within a short time-frame after EtOH exposure (i.e., during intoxication), whereas conditioning during EtOH withdrawal tends to enhance fear conditioning. The studies proposed here will therefore examine the phase-specific influence of EtOH on fear conditioning. Our central hypothesis is that phase-specific expression of cytokines in the basolateral amygdala (BLA) produce opposing actions on BLA excitability and subsequent fear conditioning. These studies will also examine long-term adaptations in neuroimmune function and resultant consequences following adolescent CIE. In this way, the proposed studies will be among the first to examine how phase-specific, EtOH-induced cytokine expression translates into age-specific, cognitive and behavioral outcomes of early EtOH exposure.