Charles V. Vorhees, PhD - US grants

Both advanced maternal age and multiparity are factors which place the human conceptus at risk. It has been shown recently that mothers of fetal alcohol syndrome (FAS) children have had more pregnancies and are older than mothers who give birth to normal children. The present study will examine both maternal age and parity in a rodent model of FAS. Our hypothesis is that old dams with a high parity history will produce the most severely affected offspring. It is the aim of this study to provide information on the effects of prenatal alcohol and maternal age, maternal parity, and the combination of both age and parity on pregnancy outcome and offspring development. Pregnancy outcome will be assessed by measuring maternal weight, resorptions, malformations (external), sex ratio, and birth weights. Offspring development will be assessed by measuring growth, mortality, and behavior. Offspring behavior that will be measured will be olfaction, activity and learning. This information combined with knowledge of the mother's drinking pattern may provide insight into the potential risks that produce an FAS child.

This research is to develop and evaluate in an animal model a new treatment for maternal phenylketonuria (PKU). Maternal PKU produces a high incidence of children born with mental retardation (up to 92%), growth retardation, microcephaly and congenetal malformations, particularly cardiac defects. Dietary treatment of PKU women during pregnancy by restricting phenylalanine intake has not veen very successful in preventing the intrauterine damage. In recent years, we have been developing a new treatment for older PKU individuals that involves the administration of the amino acids valine, isoleucine and leucine (VIL). The treatment was developed because of the difficulty in being able to adequately restrict dietary phenylalanine in PKU adults whose protein requirements are low compared to children. VIL is able to reduce CSF phenylalanine and improve symptoms of acute phenylalanine toxicity in adults with PKU. We now propose to test the feasibility of the VIL treatment in cases of maternal PKU. As a first step, studies in rats will be undertaken using an animal model of PKU involving the administration of L-phenylalanine and p-chlorophenylalanine. We will use the PKU-inducing diet to induce malformations, growth retardation and behavioral impairments in utero in pregnant rats. Once the timing of administration, exact dosage of phenylalanine and p-chlorophenylalanine are established, we will test the ability of varying doses of VIL to counteract the adverse effects of the PKU diet. We will use a range of control procedures to rule out explanations for our findings due to inadequate food or fluid intake, the influence of p-chlorophenylalanine or phenylalanine separately, VIL alone, and for the postnatal studies, of postnatal maternal influences. The data obtained will lay the groundwork for human trials of the VIL treatment in pregnant PKU women.

Both advanced maternal age and multiparity are factors which place the human conceptus at risk. It has been shown recently that mothers of fetal alcohol syndrome (FAS) children have had more pregnancies and are older than mothers who give birth to normal children. The present study will examine both maternal age and parity in a rodent model of FAS. Our hypothesis is that old dams with a high parity history will produce the most severely affected offspring. It is the aim of this study to provide information on the effects of prenatal alcohol and maternal age, maternal parity, and the combination of both age and parity on pregnancy outcome and offspring development. Pregnancy outcome will be assessed by measuring maternal weight, resorptions, malformations (external), sex ratio, and birth weights. Offspring development will be assessed by measuring growth, mortality, and behavior. Offspring behavior that will be measured will be olfaction, activity and learning. This information combined with knowledge of the mother's drinking pattern may provide insight into the potential risks that produce an FAS child.

Up to half of the 3.5 million babies born annually in the United States are exposed to diagnostic ultrasound in utero. No adverse effects, and many benefits, have accrued to ultrasound's proper use during pregnancy, but these are lingering concerns about its possible effects on subtle aspects of intrauterine brain ontogeny and subsequent behavioral and intellectual development. The goal of this research is to evaluate the possible biobehavioral effects of ultrasound on the developing central nervous system by exposure of rat embryos and fetuses to ultrasound and evaluating the offspring with tests recently developed here and elsewhere to detect behavioral teratogenicity. The structural and ponderal teratogenicity of ultrasound in separate groups identically exposed will also be evaluated. A unique feature of the proposal is that we will insonate rats without subjecting them to long periods of anesthesia or other forced restraint during gestation by using a behavioral approach in which we will train the rats to voluntarily remain stationary while propping themselves in a water vessel. We already have data that this can be done successfully. In the course of 5 years we propose to examine the dose-response effects of continuous wave (yr-01) and pulsed wave (yr-02) ultrasound (4 levels of each type, 2-3 of which are in the clinical range). Exposure will be during most of pregnancy (postimplantation to parturition). The structural, ponderal, neurological, and behavioral development of the offspring will be evaluated in detail. We will also determine if ultrasound produces any residual maternal effects lasting until after delivery that might affect the dam's rearing of her young (yr-03). Finally, we will assess whether ultrasound at one of four very specific stages of gestation has any effects on postnatal neurological, behavioral, and intellectual development (yr-04 & 05). The stages of exposure for this last 2-year study will be preimplantation, early-mid organogenesis, late organogenesis, and fetogenesis, all stages that are clinically relevant.

DESCRIPTION: (Applicant's Abstract) Effects on children from intrauterine exposure to methamphetamine (MA) have been the focus of concern because although taken less frequently than cocaine its popularity in smoked form has increased in several regions of the U.S. In humans, MA has not been linked to congenital malformations, but preliminary indications of neurotoxicity have been reported. During the last funding period, following developmental MA treatment, we found persistent augmented acoustic startle reflexes, reduced nucleus accumbens serotonin, and most unexpectedly, spatial learning deficits. The focus of this application is to extend these findings in order to: (1) test the specificity of the spatial learning deficits by comparing its effects to nonspatial (proximal cue) learning, (2) test for its effects on working memory, (3) determine its effects on spatial discrimination and compare that to nonspatial discrimination, (4) investigate cholinergic and noradrenergic pathways to the hippocampus in relation to the spatial learning effects, noradrenergic innervation in the amygdala in relation to the startle effects, and changes in NMDA receptors in the hippocampus as another possible correlate of spatial learning deficits, (5) resolve several technical issues related to: dose rate (4 vs. 2 injection schedules), internal dose (pharmacokinetics), inclusion of undernourished controls in neonatal studies, and fostering/crossfostering controls in prenatal studies. The learning/memory procedures will be conducted in a Morris water maze. The Porsolt swim test will be used to determine if MA offspring have altered stress responsiveness similar to that reported after prenatal cocaine treatment. Reactivity procedures will be conducted using basic and fear-potentiated acoustic startle. Four exposure periods (2 prenatal and 2 neonatal) were chosen to span distinct stages of brain development (analogous to first, second, and early third human trimester brain development). The results will define the specificity of MA's developmental effects on spatial learning and startle, and begin the search for neurochemical correlates of these functional changes. The findings may assist in understanding the long-term risks to children exposed in utero to MA.

DESCRIPTION: (Applicant's Abstract) We have found spatial learning deficits after exposure to methamphetamine (MA) and MDMA on postnatal days 11-20. Based on comparative CNS ontogeny, this period in rats is analogous to human 2nd-3rd trimester development. Exposures from weaning to puberty in rodents (equivalent to childhood to adolescent development) are poorly understood, despite the fact that children and adolescents sometimes abuse stimulants. Neurochemically, we have found significant changes in dopamine and GFAP between postnatal day 20 and 60 in response to MA, ages spanning the so-called periadolescent period of rodent development. Based on these data, it appears that there are changes occurring during these stages of brain development which could affect MA-induced neurotoxicity. The purpose of this application is to investigate the cognitive effects of exposure to MA to determine if there are later periods of vulnerability. Assessments will include the Morris water maze test of spatial navigation, tests of working and sequential learning, and conditioned place preference (CPP) behavior. Effects on hippocampal norepinephrine will be investigated based on new data showing that MDMA increases NE after preweaning exposure. Split-litter-by-exposure-age experiments will first determine the most vulnerable ages for learning and CPP effects; these will be followed by split-litter dose-response experiments for those periods showing the largest effects and will investigate effects on working memory. Exposure periods will be postnatal days 21-30, 31-40, 41-50, and 51-60. A final experiment to differentiate the previously found preweaning vulnerable period will be investigated after treatment on postnatal days 11-15 or 16-20. Experiments will begin with MA treatment and proceed to treatment with MDMA depending on the outcome of the MA experiments. NIDA has identified an important gap in our understanding of the effects of drugs of abuse on cognitive development and its neurobiological substrates and this application will develop data to fill this gap in an animal model of MA and MDMA-induced neurotoxicity."

Methamphetamine (MA) abuse has risen in recent years and in some regions is used more than cocaine. Despite this, little is known about its prenatal effects. During the first funding period we established that a third trimester equivalent exposure to high doses of MA in rats results in acoustic startle hyperreactivity and deficits and deficits in learning the Morris maze hidden platform test of spatial navigation. During the most recent funding, we extended these findings and determined that the spatial learning deficit is replicable, generalizable (found in 3 strains), obtained over a range of doses, specific to spatial navigation, and affects reference but not working memory. We also obtained pharmacokinetic data showing that P1-10 and P11-20 treatment produce similar absorption, peak and elimination patterns despite the fact that only P11-20 leads to spatial learning impairments. We also found that MA at these developmental ages does not induce hyperthermia or alter brain monoamines, but that it does dramatically increase plasma corticosterone. We now propose to pursue these findings with aims to determine the effects of MA at different developmental stages . For the third trimester equivalent exposure period we plan to examine MA's: (1) longitudinal effects on spatial learning, (2) strategic and motivational specificity, (3) dose-response effects, (4) critical period effects, (5) conditional place preference effects, (6) glucocorticoid and glucocorticoid receptor effects, and (7) NMDA receptor effects. For first and second trimester equivalent exposure periods we will determine MA's effects on: (8) spatial learning and memory (with controls material influences), and (8) glucocorticoids. These studies will advance our understanding of MA's effects on the developing brain neurochemically and functionally, especially MA's cognitive effects, and provide new information that may ultimately assist in extrapolating these findings to the risk infants and children may encounter when their mother's abuse MA during pregnancy.

3,4-Methylenedioxymethamphetamine (MDMA) is an amphetamine analog that possesses both stimulant and hallucinogenic properties. In recently completed preliminary experiments we have identified neurobehavioral effects following developmental MDMA exposure. Specifically, neonatal MDMA administration (analogous to late in utero stages of human development) results in long-lasting deficits in spatial and sequential water maze learning and memory. The largest effects occurred after MDMA treatment on postnatal days 11-20, with smaller effects after treatment on days 1-10. In a follow up experiment completed since the last submission, we replicated these effects in a between-litter design and showed that the effects were not the result of undernutrition or saline-injection stress occurring during critical stages of development. The primary focus of this proposal is to characterize those aspects of learning and memory that are disrupted following developmental MDMA exposure and to identify the endocrine changes that are associated with these effects. To accomplish this the following specific aims will be investigated: (la) Test the hypothesis that deficits in the Morris water maze following developmental MDMA exposure result from disruptions in working memory using both appetitive (Olton maze) and aversively motivated (working memory version of the Morris maze) approaches and (lb) conduct pharmacokinetic studies on neonatal rats exposed to MDMA at the two exposure stages of interest to determine if there are developmental differences in brain or plasma concentrations of drug that might explain the stage-specific cognitive deficits we have found. (2) Determine the time-dependent onset and long-term effects of the induced cognitive deficits by examining animals at different developmental stages after treatment with MDMA. (3) Determine if the adult cognitive effects could be associated with HPA axis changes as reflected by alterations in plasma corticosterone and ACTH, or hypothalamic CRF; or by alterations in fearfulness as measured in the elevated plus maze (uncontaminated by changes in locomotor activity levels). Further, we will determine whether, in a test of distracter cues on Morris maze, alterations in attention may contribute to the learning and memory changes. Changes in this revised application include the addition o multiple dose groups to all of the behavioral experiments and to the pharmacokinetic and endocrine experiments, elimination of monoamine determinations, and reducing, the scope of work on NMDA and NE receptor binding to pilot studies. The expanded dose-response experiments (including counter-balancing for cross-task effects) will be the first of their kind to demonstrate that developmental MDMA exposure induces long-term learning and memory impairments. Such data will help identify risks associated with the prenatal use of MDMA on brain and cognitive development.

DESCRIPTION (provided by applicant) The goal of this training program is to educate scientists who will contribute to the fight to prevent birth defects (congenital malformations and neurobehavioral abnormalities). The approach taken is to view birth defects as molecular perturbations of developmental pathways. Graduate students supported by this grant in the Molecular and Developmental Biology (MDB) Graduate Program take classes in developmental biology, embryology, molecular genetics, biochemistry, cell biology, and teratology and for some also in neuroscience and biostatistics. The molecular focus is carried throughout the program, including seminar and journal clubs, and in laboratory experiences. All laboratories in the program use molecular techniques and many also use cellular and systems methodologies. Trainees from this program have been very successful, moving on to academic, industry and government research positions in developmental toxicology, teratology, and/or developmental neurotoxicology. There is a national shortage of scientists trained in these areas and demand for our graduates remains strong. Our graduates have proven to be well prepared causing employers to return to the investigators for more of their trainees. Accordingly, request is made to train four pre-doctoral students and three postdoctoral fellows. This is the same number of pre-doctoral positions and an increase of one postdoctoral position compared to the last five years. The request for three postdoctoral positions is the same as was funded during the preceding cycle (10 to 5 years ago) and the same as was approved for funding 5 years ago. Since that time demand for scientists trained in these areas has increased and the number of applicants for the postdoctoral positions has also increased. In response, the investigators have expanded the training faculty and seek reinstatement to the prior level of three postdoctoral positions. The duration of training for postdoctoral fellows is three years and for graduate students, five years. It should be noted that recent changes to the MDB curriculum have the potential to reduce the time to earn the Ph.D. degree to 4.5 years. In addition, the contexts in which this training grant operate, i.e., the MDB program and the Children?s Hospital Research Foundation, are undergoing major expansions that bode well for even greater future success of this training grant in the next five years.

DESCRIPTION (provided by applicant): 3,4-Methylenedioxymethamphetamine (MDMA) abuse is a serious health problem yet little is known about its effects on developing brain. We established that neonatal rats administered MDMA (a model of 3rd trimester exposure) causes long-term path integration and spatial learning and memory impairments after P11-20 (but not P1-10) exposure, while sparing cued and working memory. New data show that exposed offspring have altered expression of CAPON, PSD95, nNOS, and NMDA-NR1 (all members of the NMDA receptor complex). This treatment also causes large reductions in 5-HT and sustained release of corticosterone (CORT). P11-20 overlaps the stress hyporesponseive period (SHRP, P4-15). We hypothesize that MDMA treatment that begins during the SHRP triggers a unique cascade of events beginning with overactivation of the stress response pathway (release of CRF, ACTH, CORT) but in which normal feedback mechanisms fail to operate correctly. The resulting prolonged CORT release alone or combined with the concomitant 5-HT reductions lead to changes in CNS organization and long-term cognitive deficits. Specific aims to test this hypothesis are: (1a) Determine the critical period for MDMA-induced long-term cognitive and NMDA receptor complex effects using treatment intervals before, during and after the SHRP and for changes in response to stress. (1b) Compare the critical period from Aim 1a with a non-critical period for short-term effects on the HPA axis and brain 5-HT. (2) Test the role of HPA axis changes in the long-term effects using a new technique we developed involving adrenalectomy combined with adrenal alloengraftment to temporarily interrupt HPA responses with later restoration of basal CORT function. (3) Test 5-HT involvement by blocking MDMA-induced 5-HT reduction using SSRI pretreatment and test for prevention of cognitive deficits. (4) Determine changes .in nNOS, NMDA-NR1, PSD-95 and CAPON and related proteins in animals not tested behaviorally to ensure that these changes are not experience-dependent. The present model of developmental MDMA exposure is the first to establish induction of cognitve deficits and is the first step in our long range objective to ultimately test the effects of MDMA after exposure during other (earlier) stages of brain development.

[unreadable] DESCRIPTION (provided by applicant): Environmental exposures to polychlorinated biphenyls (PCBs) are known in humans as well as lab animals to cause immunosuppression, thyroid disease, endocrine disruption, and damage to the central nervous system. Not all humans or laboratory animals respond similarly to the same dose -indicating interindividual genetic differences. In rodents to elicit these pathologies, planar PCBs must bind to, and activate, the aryl hydrocarbon receptor (AHR). Despite this overriding role for planar-PCB-mediated AHR activation in toxicity, the AHR up-regulates CYP1A2, which in liver sequesters and protects distant tissues against planar PCBs. Both the AHR and CYP1A2 are polymorphic in humans: the AHR exhibits >12-fold differences in ligand-binding affinity; liver basal CYP1A2 shows >60-fold differences in subjects having no known exposure to inducers. With regard to fetal exposure, our studies in mice demonstrate that risk of birth defects by planar TCDD depends on the high-affinity AHR and is also greatly increased in fetuses carried by dams that lack CYP1A2. PCBs represent mixtures having many dozens of different congeners; which congener is toxic, and the rates of uptake, metabolism and excretion are difficult to determine in humans, and most studies in lab animals look at a single congener. We have studied mice with the high- (Ahrb, B6) vs low- (B6.D2-Ahrd) affinity AHR, and with or without the Cyp1a2 gene. Using these mice, we hypothesize that Ahrb fetuses carried by Cyp1a2(-/-) dams will be most susceptible, and Ahrd fetuses carried by Cyp1a2(+/+) dams most resistant, to deficits in learning, memory, and other behaviors caused by planar PCBs. For the funding period, we propose to: [1] determine tissue distribution of each of eight PCB congeners (most relevant to humans) given as a mixture -comparing B6 vs B6.D2-Ahrd, and Cyp1a2(+/+) vs Cyp1a2(-/-) dams and their offspring; and [2] evaluate the in utero and lactational effects of this orally administered PCB mixture on learning, memory, and other behaviors in offspring of these treated dams, starting at postnatal day 60. These studies will define the impact of a fetal basis for adult disease. The Ahr and Cyp1a2 genotypes in these mice represent the extremes for variability of these two genes in the human population. There exist genetic differences in mouse (and human) populations, which represent a gradient of at-risk individuals. Project Narrative: Polychlorinated biphenyls (PCBs) are widespread persistent organic pollutants linked to numerous human health problems, including learning and memory deficits in children of exposed mothers. The Ahr and Cyp1a2 genotypes in our mouse models represent the extremes for variability in these two genes in the human population, and both genes likely play a role in susceptibility following PCB exposure. These studies will define the fetal basis for adult disease and help to identify individuals at greatest risk of PCB-induced neurotoxicity. [unreadable] [unreadable] [unreadable] [unreadable]

This request is for support to help defray the costs of the Annual Meeting of the Neurobehavioral Teratology Society (NBTS) for 5 years. The annual meeting of NBTS highlights new data and theoretical constructs concerning neurobehavioral teratology and the developmental origins of CNS disorders that result from developmental exposure to drugs, toxic substances, environmental contaminants (including food contaminants), stressors, genetic mutations, epigenetic changes, and the interaction of these factors. Effects in humans, animal models, and neural studies of mechanisms are presented. Issues of construct and translational validity are considered, as well as developments concerning the mechanisms of how drugs and other chemicals affect brain and behavior. The 2013 meeting features topics on advances in methods, advances in the science of developmental disorders, and brings forward areas (e.g., schizophrenia) as issues of importance since recent data suggest that neuropsychiatric disorders may have embryonic and fetal origins. The meeting brings together developmental neuroscientists, developmental neurotoxicologists, developmental psychologists, developmental epidemiologists, teratologists, developmental toxicologists, medical geneticists, physicians, genetic councilors, and developmental environmental scientists from academia, industry, and government. NBTS meets with the Teratology Society (TS) and the Organization of Teratology Information Specialists (OTIS) producing synergy. Aim 1: Support Conference Travel Awards for 2 graduate students, postdoctoral fellows, advanced undergraduates or research assistants to present their data with waiver of registration fee and discounted room rates. Conference Awards are $700 each. The Society matches this by supporting 2 conference travel awards from its dues for a total of 4 awards per year. Aim 2: Support 1 New Investigator Award per year. The Society waives the recipient's registration fee and pays for travel and meeting-related expenses ($1100 coming from this grant and the balance ($2000-2500) from the Society. Aim 3: Provide support for travel expenses for non-member speakers. NBTS typically invites 8-10 non-member speakers per year. We request $2,500 toward this cost. The 2013 outside speaker estimated costs will be ~$15,000. Hence, the $2,500 contribution requested will be partial support to help the Society maintain the involvement of high quality non-member speakers.

DESCRIPTION (provided by applicant): Attention Deficit Hyperactivity Disorder (ADHD) affects 5.4 million children at a cost >$72 billion/year, yet its etiology is poorly understood. A new gene for an orphan receptor has been identified that conveys high risk for ADHD: Latrophilin-3 (LPHN3). We have a new Lphn3 knock-out (KO) rat. This rat shows hyperactivity and hyper-reactivity. But ADHD also affects attention and impulse control. Objective-1 will test Lphn3 KO rats for attention and inhibitory control. ADHD is treated with stimulants, which increase attention and reduce distractibility and activity. Objective-2 is to test the effects of ADHD medications in Lphn3 KO rats. Effective ADHD medications increase dopamine and norepinephrine signaling (and some increase 5HT) by acting on monoamine reuptake and release processes or receptors. Objective-3 is to determine the effects of Lphn3 disruption on monoamines, transporters, and receptors. The overall purpose is to identify the contribution of Lphn3 to the behavioral and neurochemical phenotype of ADHD. The proposed experiments will generate essential data for how Lphn3 contributes to each of the symptoms and neurochemical changes associated with ADHD. Lphn3 could become a novel target for development of drugs that modulate Lphn3 function or its binding partner (Flrt3) to alleviate the underlying deficit in ADHD. As such this represents innovative research on a cause of ADHD. We are the only group with the Lphn3 KO rat and pilot phenotypic data. Therefore, we are uniquely position to pursue this model. Specific Aim-1 will determine the ADHD-like phenotype of Lphn3 KO rats. The role of Lphn3 loss of function in extended locomotor activity, sensorimotor gating, cognitive flexibility/attention (set shifting), attention/flexibility (continuous performance test), working memory, and response to ADHD medications will be assessed. The data will demonstrate the range and severity of ADHD phenotype. Specific Aim-2 will elucidate the neurochemical changes in Lphn3 KO rats. Exp-2a: KO and WT rats will be compared by immunohistochemistry for changes in monoamines implicated in ADHD (dopamine (by tyrosine hydroxylase), 5HT, and dopamine beta hydroxylase (for NE), and as controls GAD67 (for GABA) and NMDA-NR1 (for glutamate). Stains for each transporter will be performed. Because the dopamine D4 and 5HT2B receptors have been implicated in ADHD they will also be assessed. Exp-2b: Once histological regions of change are identified, other animals will have these regions analyzed by Western blot. Exp-2c: Once Westerns from Aim-2b identify significant changes, this will be used to guide which markers will be analyzed in Aim-2c in animals after behavioral testing; then correlations between neurochemical and behavioral data will be performed. In addition, RT-PCR for Flrt3 will be performed to determine it is altered by the Lphn3 mutation. These experiments will materially advance the field of ADHD research.

DESCRIPTION (provided by applicant): The objective of our application is to determine if exposure to polychlorinated biphenyls (PCBs) causes epigenetic transgenerational effects. There is extensive evidence that early life exposure to environmental chemicals can lead to disease outcomes in adult life. The negative effects of these exposures are believed to be reset in each generation, such that subsequent generations are unaffected by the exposure history of their parents and grandparents. However, evidence challenging this notion is growing, suggesting that in certain cases, environmental exposures affect multiple generations removed from the original insult, causing far- reaching consequences. It is still controversial whether thi phenomenon, known as transgenerational inheritance of acquired traits, occurs in mammals, though exposure to specific diets or nutritional factors, maternal stress, and certain environmental chemicals have all been reported to induce phenotypic changes observed at least two generations after exposure. We have shown that progeny of dams treated in utero with a mixture of coplanar and non-coplanar PCBs that simulates human exposure show cognitive deficits as adults. Other experimental evidence in rodents and primates strongly suggests a transgenerational impact of in utero exposure as the cause of abnormal fetal neurodevelopment and later childhood cognitive function. There is also compelling evidence of an association of PCB exposure with decreases in neurological function in infants and children, including Attention Deficit Hyperactivity Disorder. Similarly, maternal exposure during pregnancy to PCBs and other halogenated hydrocarbons has been associated with a higher incidence of congenital heart defects in the progeny and cardiac insufficiency later in life, linking fetal and adult cardiovascular disease. It remains to be determined if these health effects are caused by transgenerational or direct toxicity because studies have not been carried out to the F3 generation. In this application, we propose to address the hypothesis that exposure to PCBs during a critical window of development causes heritable transgenerational phenotypic changes in CNS and cardiac physiology associated with changes in DNA methylation patterns responsible for altered chromatin conformation and subsequent dysregulation of gene expression, detectable at least up to the F3 generation. In three Specific Aims, we will determine whether developmental PCB exposure during the time of germ-line specification will change neural or cardiac phenotypes transgenerationally and whether these changes will be associated with corresponding changes in the germ-line epigenome and affected tissue epigenomes and transcriptomes. Successful demonstration of transgenerational inheritance in response to in utero PCB exposure will transform our understanding of the health effects of these ubiquitous environmental pollutants.

This application is for support of the Annual Meeting of the Developmental Neurotoxicology Society (DNTS). The annual meeting of DNTS highlights new data and theoretical constructs concerning developmental neurotoxicology and the developmental origins of adult CNS disorders that result from developmental exposure to drugs, environmental agents, pesticides, and contaminants (including food contaminants), stressors, genetic mutations, epigenetic changes, and interactions among them. Effects on humans and experimental animals are used, and studies of cellular, molecular, physiological, and behavioral effects/mechanisms are presented. Issues of construct and translational validity are considered, as well as developments concerning the mechanisms of how drugs and chemicals affect brain and behavioral ontogeny. The 2018 meeting featured topics on methods, advances in the science of developmental disorders, and problems of American society, such as tobacco, marijuana, opioid addiction, and medications to treat addiction during pregnancy, effects of environmental agents and potential causes of children?s neurobehavioral disorders. The meeting brings together developmental neuroscientists, developmental neurotoxicologists, developmental psychologists, developmental epidemiologists, teratologists, developmental toxicologists, medical geneticists, physicians, genetic councilors, and environmental scientists from academia, industry, and government. DNTS meets with the Teratology Society (TS) and the Organization of Teratology Information Specialists (OTIS) cooperatively. Aim 1: Support Conference Travel Awards for 4 trainees (graduate students, postdoctoral fellows, undergraduates or research assistants) to present their newest data. Conference Awards are $700 each. The Society matches this by supporting 4 additional conference travel awards. Aim 2: Support (50%) for the Patricia Rodier Mid-Career Award jointly with TS. Aim 3: Provide partial support for travel expenses for non- member, invited speakers. DNTS typically invites 8-10 non-member speakers per year. We request $2,500 toward this cost. A recent meeting cost for outside speakers was $15,000. Hence, $2,500 requested will be a fraction of the total cost of outside speakers but is critical to the vibrancy of the Society?s Annual Meetings. Five years of support is requested at $6,000/year.

Attention Deficit Hyperactivity Disorder (ADHD) is the most prevalent neurodevelopmental psychiatric disorder (9.4% prevalence in children; 4.4% in adults) and is polygenic. A novel gene associated with ADHD is Latrophilin-3 found in striatum, hippocampus, cerebellum, prefrontal cortex (PFC), and amygdala. In humans, there are 21 variants of LPHN3 associated with ADHD. Some pesticides may interact with ADHD genetic risk factors to trigger or exacerbate the symptoms. We found that the common pyrethroid, deltamethrin (DLM), administered prior to weaning in rats causes long-term behavioral, neurochemical, and electrophysiological effects. We developed the first KO rats of Lphn3. Lphn3 KO rats are hyperactive, hyper-reactive to startle stimuli, and cognitively impaired. This PAR-19-386 ?Environmental Risks for Psychiatric Disorders: Biological Basis of Pathophysiology? seeks models that will elucidate Gene x Environment interactions related to neuropsychiatric disorders, such as ADHD. We hypothesize that Lphn3-/- and Lphn3+/- rats will interact with DLM (Type II pyrethroid) or permethrin (PRM, Type I pyrethroid) to exacerbate an ADHD-like phenotype. Specific Aim 1: Determine the effects of DLM in Lphn3-/-, Lphn3+/-, and wildtype (WT) rats on activity, reactivity, learning and memory (L&M), dopamine (DA) and NMDA markers, and apoptosis. Aim-1a: Compare WT rats with Lphn3-/- and Lphn3+/- rats administered 0, 0.5, or 2.0 mg/kg DLM from P3-20 for changes in activity, acoustic and tactile startle (including prepulse inhibition (PPI)) egocentric, allocentric, and working L&M, and for changes in DA and NMDA-R markers in various brain regions, including markers for programmed cell death. Aim-1b, neurochemical outcomes in rats not behaviorally tested. Specific Aim 2: Determine the effects of PRM in Lphn3-/-, Lphn3+/- rats vs. WT rats on the outcomes used in Aim-1. Aim-2a: Same as Aim-1a with PRM. Aim-2b: Same as Aim-1b with PRM. Specific Aim 3: Determine the effects of DLM in adult Lphn3- /-, Lphn3+/- rats vs. WT rats. Aim-3a: same outcomes as in Aim-1a. Adults with ADHD are an understudied and a population susceptible for higher exposure to pyrethroids from occupational exposure, making Aims 3 and 4 important. Aim-3b: Same as Aim-1b in adult rats. Specific Aim 4: Determine the effects of PRM in adult Lphn3-/-, Lphn3+/- rats vs. WT rats. Aim-4a: Same outcomes used in Aim-1a. Aim-4b: Same as Aim-1b in adult rats not behaviorally tested. Impact: ADHD interferes with normal development, costs billions to treat and manage, yet we know little about environmental contributions to those with ADHD. Insecticides are suspected in ADHD but such interactions between gene and environment are not established. Lphn3-/- and Lphn3+/- rats represent a novel approach to probing the effects of exposure to pyrethroids using a known ADHD genetic susceptibility. The model will shed new light on how a gene known to be associated with ADHD affects the behavioral and biochemical effects of prototypical pyrethroids. Interaction data can be used for risk assessment and help provide safeguards against pyrethroid exposure for those with ADHD.