Prescribed opioids are the most common analgesic used for alleviating acute and chronic pain. Despite this positive attribute, opioids are also highly abused drugs that can lead to tolerance and dependence. This abuse has caused a dramatic increase in opioid overdose-related deaths over the past couple of decades, deeming it a crisis. However, cessation of opioid use in tolerant individuals who wish to detoxify can precipitate severe withdrawal symptoms, often leading to relapse in order to avoid experiencing these negative symptoms. In recent studies, modulation of neuropathic and neuroinflammatory responses have been linked to withdrawal symptoms. As a result, we hypothesized that microglia, the resident immune cell of the central nervous system, serve as a potential target for withdrawal treatment. In order to test this, we reduced microglial activity by inhibiting the purinergic signaling pathway. This was achieved by first exposing mice to escalating doses of fentanyl over the course of a few days to create tolerance. Then, we administered clopidogrel, a selective antagonist of the P2Y12 receptors which are expressed in microglia, before inducing withdrawal using naloxone. Subsequently, in order to quantify whether inhibition of microglial P2Y12 receptors mitigated naloxone-precipitated withdrawal in fentanyl-tolerant mice, we measured avoidance of the withdrawal context with the conditioned place aversion (CPA) test, and evaluated somatic signs of withdrawal with EthoVision video analysis. Avoidance of the negative emotional and physical symptoms of withdrawal is a key driver of relapse, therefore the results from this experiment can provide prospective molecular pathways to target for future studies in treating opioid withdrawal symptoms. Reducing the severity of withdrawal would thus allow ease in discontinuing opioid use and diminish relapse.

Pituitary adenylate cyclase-activating polypeptide (PACAP) is an excitatory neuropeptide which has been associated previously with stress, fear, and post-traumatic stress disorder (PTSD). One region that strongly expresses Adcyap1, the gene encoding PACAP, is the lateral habenula (LHb), a node of stress in the brain. Studying PACAP and its role within the LHb provide insight into the aid and treatment of a variety of stress disorders by understanding the mechanisms of these conditions. In order to investigate the function of PACAP within the LHb we used a 2x2 experimental design. We injected a virus expressing either a Cre-inducible excitatory Designer Receptor Exclusively Activated by Designer Drugs (DREADD) hM3Dq and RiboTag or Cre-inducible RiboTag alone into the LHb of Adcyap1-2a-Cre mice to activate and quantify gene expression in these neurons specifically. Mice were injected with either the DREADD-specific drug clozapine-N-oxide (CNO) or vehicle just prior to contextual fear conditioning, a behavioral procedure in which mice are placed in a novel chamber and given repeated foot shocks in order to elicit a fear memory. The following day, mice were placed in the same chamber without CNO and their time spent freezing indicated the strength of their fear memory. We hypothesized that mice which have their LHb PACAP neurons activated will have increased time spent freezing within the contextual fear chamber, indicating they have a stronger fear memory than the control groups. This study could shed light on the mechanisms of PTSD and other stress disorders.

Opioid abuse leads to over 40,000 annual deaths in the United States; even more individuals are impacted by anticipatory withdrawal anxiety and subsequent treatment avoidance. Despite the magnitude of this issue, there is a lack of effective treatments that address opioid dependence and withdrawal. Molecular responses to opioids have traditionally been linked to neuronal activity, but recent literature suggests that microglia also play a role in opioid addiction. Recent experiments we conducted reveal that opioid dependence and withdrawal have inverse effects on the microglia translatome, with morphine treatment correlating to decreased gene expression and withdrawal correlating to increased gene expression. We found a dramatic change in genes relating to cyclic AMP signaling during withdrawal, which has been shown to modulate microglia motility and potentially their interactions with nearby neurons. From this, we sought to further investigate the molecular basis of microglia morphology during opioid tolerance and withdrawal. To do so, we constructed four experimental groups consisting of mice who received saline followed by saline, saline followed by naloxone, morphine followed by saline, and morphine followed by naloxone. After obtaining the mouse brains through perfusion, we took sections of the striatum. In order to visualize and quantify microglia morphology, we performed 1ba1 immunohistochemistry to stain the slices, then imaged mounted slices on a confocal microscope to acquire confocal stacks of the striatum. This was followed by 3D reconstruction of individual microglia for analysis using 3DMorph software. The results of this experiment are a step towards clarification of molecular mechanisms behind opioid dependence and withdrawal for future work on mitigating the effects of opioid addiction. Alleviating withdrawal symptoms through translational research would allow users to more easily cease opioid use and therefore reduce opioid abuse mortality.

Despite research supporting the efficacy of certain mental health practices, many mental health care providers in community mental health institutions are not utilizing evidence-based practices (EBPs) consistently as their method towards patients receiving mental health care. This is a problem because the patients in need do not benefit from therapies intended to help them. Clinical supervisors (clinicians who provide clinical direction and guidance to less experienced providers) play an important role in how often and how effective clinicians are at implementing EBP’s in their treatments. The purpose of our project is to examine the feasibility and self-reported usefulness of providing case-based consultation to clinical supervisors in supporting the implementation of EBP. We expect that clinical supervisors will report that they benefited from EBP case-based consultations by having an increase in confidence and competence in supervising mental health care providers in their efforts to use EBP. So far, we have successfully recruited 9 clinical supervisors from 9 community mental health organizations. Clinical supervisors completed a pre-survey that tested their comfortability in supervising mental health care providers implementing EBP. In addition to gathering quantitative data via the survey, we are also analyzing qualitative data from transcripts of the consultation calls with the clinical supervisors. After the 6 consultation calls, we expect that supervisors will rate the calls as high in usefulness. We also plan to learn about any barriers or challenges that the clinical supervisors experienced. With the data that we have collected during this pilot test, we hope to conduct further research testing if EBP implementation to clinical supervisors through case-based consultations creates a downstream effect by resulting in an increased level of competency and comfortability for clinicians.  

Autism Spectrum Disorder (ASD) is a disorder characterized by social and communicative impairments. Children with Autism Spectrum Disorder, particularly those who are older, display internalizing behaviors more frequently than typically developing children. These behaviors are mainly characterized by negative feelings and occur at higher rates in females than males in children with ASD, however, the specific rates in each subgroup, such as anxiety, depression, or somatic complaints are less understood. This study will aim to explore the gender differences and the impact of the following factors on internalizing behaviors in children with ASD: (1) Participant medication use (for behavioral issues), (2) symptoms of depression/anxiety in parents of participants, (3) environmental factors such as family composition and household income. 161 participants, aged 8-18 (90 boys, 71 girls) from the ACE GENDAAR network, a four site NIH funded project investigating gender differences in children with ASD, were included in the study. Additionally, all participants met ASD criteria on ADOS-2 and via the ADI-R. Parents of participants completed the Child Behavior Checklist (CBCL), answering questions about their child’s internalizing behaviors. Parents also completed the ACE Medical History (medication use) and the ACE Demographics (environmental factors) questionnaires. We hypothesize that girls with ASD will score higher (worse) across all internalizing behavior symptom categories. We also expect to see correlations between the use of medication by participants, depression/anxiety reported in parents of participants and internalizing behavior symptoms. The data from this study will not only improve our understanding of specific internalizing behaviors associated with girls and boys with ASD but will also provide insight into the effects of various medical and environmental factors affecting children with ASD.

Autism Spectrum Disorder (ASD) is a developmental and neurological disorder characterized by difficulties in communication and social domains. Previous research highlights that involvement in organized community activity has shown to have positive effects on social and emotional adjustment in youths (Bohnert et al., 2019). Meeting early language milestones strongly indicates positive language development in individuals with ASD. Better early language development may foster interactions with peers, promote a sense of belonging, and may make it easier for parents and children with ASD to participate in community activities. 
This study explores the relationship between early language milestones (age at first words, age at 3-word phrases), current social milestones and current language level. 105 youth (f=42; m=63), ages 8-17 years with from the ACE GENDAAR network, a four-site NIH funded project investigating gender differences in children with ASD, were included in the study. ASD diagnosis was confirmed via standardized measures of autism symptoms (ADOS-2, ADI). All included participants completed the DAS-II to measure cognitive skills, with a score of ≥70 on the Verbal domain. Parents completed the ACE medical history and ADI, which assessed early language milestones. Parents also completed the Child Behavioral Checklist (CBCL) and rated their child’s activity involvement in sports, hobbies and group organizations and completed a measure of social adaptive skills. We hypothesize that there will be a positive correlation between meeting early language milestones, social involvement, and language development in children with ASD. That is, children with better early language, will have better later language, better adaptive skills, and will be more likely to participate in community activity. This study may help us learn more about the implications of early language development in children with ASD and how it impacts later abilities to participate in the community.     

Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder that is characterized by having persistent deficits in social interactions, as well as social communication impairments with restricted, repetitive behaviors. Past researchers have found that another characteristic of individuals with ASD tends to be higher rates of aggression compared to other developmental disabilities; a research study found that 68% of individuals with ASD in a sample of 1,380 children had a history of directed aggression. Due to the variety of measures that have been utilized to define aggression, there is a large amount of unexplored variability in the relationship between ASD and aggression. Social awareness is defined as an individual’s ability to cognitively understand reactions to different social situations and effectively modify these reactions to achieve beneficial social communication. Past research has found a strong correlation between high social responsiveness and low aggression, but this has not been widely studied in a population with ASD. The aim of this study is to explore the relationship between decreased social awareness and rate and type of displayed aggressive behavior.150 children, aged 8-11 years participated in the study, focusing on late elementary school. All participants met the Autism diagnostic criteria on the ADOS-2, a child-clinician interaction that measures a child’s social, repetitive behaviors and communication skills. Parents completed the Social Responsiveness Scale, which includes scales of social awareness and cognition, as well as standardized measures of aggression. We expect to see a negative correlation between the rate of aggression and social awareness such that children with ASD who are reported as lower in social awareness will have higher reports of aggression. This research will help to understand why aggression rates are high in this population and may inform social skills and school based interventions for children with autism aggressive behaviors.

 Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder that impairs an individual’s social, communication, and behavioral skills. These impairments can significantly impact an individual’s mental health. Puberty is a period in an adolescent’s life when they experience physical, social, and emotional changes. Adolescents are also especially vulnerable to these changes due to the potential disjunction of physical and cognitive systems maturing at different rates. Unfortunately, little is known about the effect of pubertal development on the mental health of individuals with ASD. The goal of this study was to investigate patterns in anxiety and depression rates across individuals with ASD during the stages of puberty. The study included children between the ages of 8 and 17 with a confirmed diagnosis of ASD via standardized measures of autism symptoms (ADOS-2 and ADI). Participants or their parents completed the Pubertal Developmental Scale (PDS) which asked seven sex-specific questions and provided a score of the child's physical progression through puberty. Parents also completed the Subject Medical History Form, Child Behavior Checklist (CBCL), Child & Adolescent Symptom Inventory-5 (CASI-5), a self-report questionnaire to identify signs of psychiatric disorders. We explored the relationship between states of pubertal development and prevalence of anxiety and depression between pubertal groups (pre/early/mid/late/post). We expected to find an increase in anxiety and depression rates in adolescents and individuals who are in the early-post stages of puberty relative to those who are pre-puberty. Additionally, we compared rates of anxiety and depression of children with ASD to their siblings as an environmental control. We expected anxiety and depression rates to be higher in children with ASD compared to their siblings. Investigating the relationship between pubertal development and anxiety and depression can help us better understand risks for mental health concerns in children on the autism spectrum.

Autism spectrum disorder (ASD) is a developmental disorder that causes challenges with speech and nonverbal communication, social interaction and repetitive behaviors. It is hypothesized that ASD is caused by a combination of genetic and environmental factors such that symptoms and behaviors present differently between individuals. Given variability in causes and symptoms of ASD, it is important to look at the effectiveness of treatments for individuals with specific genetic and behavioral profiles, including individuals with rare genetic mutations linked to ASD. Recommended treatments include biomedical, behavioral, speech, and occupational therapies, which can be expensive and time consuming for families. Speech-language therapies and other services can be provided through a school district or through private providers, but the effectiveness of these services for individuals with rare ASD-associated genetic mutations is currently unknown. The aim of this project is to compare satisfaction with school-based versus private speech-language therapy for individuals with mutations to DYRK1A or SCN2A, which are associated with ASD and language delay. In this study, participants included individuals with a disruptive mutation to either DYRK1A (n=28, ages 4-24 years) or to SCN2A (n=23, ages 3-21 years). Treatment history interviews will be analyzed to assess caregivers’ perceptions of treatment effectiveness. We hypothesize that there will be greater satisfaction with private speech-language therapy than with school-based services, as these services may allow for greater communication and coordination with parents. We also hypothesize that satisfaction with speech therapy will be highest in the DYRK1A group, as increased medical complications in SCN2A (e.g., severe seizures) may lead to reduced response to speech therapy. This study will contribute to better understanding of effective treatments and parents’ satisfaction with current services for individuals with rare genetic mutations associated with ASD, which may inform future treatment choices and recommendations.

Autism Spectrum Disorder (ASD) is characterized by disruptions in social, behavioral, and communication behaviors. Meeting early language milestones has been identified as a strong predictor of positive language outcomes individuals with ASD. Females, compared to males, show better early cognitive and language functioning, including high risk infants with and without ASD outcomes. Less is known about language trajectories in females with ASD, as they often make up a minority of research participants. In this study, we want to evaluate the relationship between early language milestones and youth language and communication ability in a sex balanced sample with ASD. The project included 137 youths, 60 females and 77 males, from ages 8-18 years with ASD. To assess language, parents reported from the ACE Medical History, which reports on age at first words and age at 3-word phrases which were confirmed with similar items in the ADI. The participant completed the CELF-4, with analysis focusing on the subdomains Recalling Sentences and Formulating Sentences, and the parent completed the Vineland Adaptive Behavior (Communication Domain). Our preliminary analysis demonstrated significant differences by sex in early language milestones as well as relation to later better language ability as a youth. Age at first words was related to later language, but only in females with ASD; while age at 3 words was related to later outcomes for males and females. It is important to understand how language develops different in males and females with ASD and being able to recognize risk factors at a young age for more accurate intervention.

Fear is an emotion that is triggered when an organism encounters danger or threat. Once fear is triggered, the nervous system induces many physical changes, and the brain responds accordingly to elicit defensive or fleeing behavior. Such fear responses are known to involve a critical brain region called the amygdala. One of the ways in which the amygdala mediates fear response is by secreting a neuropeptide called corticotropin-releasing factor (CRF). Our lab has previously found that CRF-expressing neurons in the central nucleus of the amygdala (CeA-CRF neurons) facilitate the acquisition of discriminatory fear responses and prevent fear extinction. In contrast, others have reported that activation of CeA-CRF neurons is reinforcing. To resolve these contradictory findings, we investigated the functional roles of CeA-CRF neurons in naïve and fear-conditioned mice. We hypothesized that activation of CeA-CRF neurons in a neutral context may produce reinforcing effects by signaling a type of salience. However, following a fearful experience these neurons may signal the aversive nature of this experience. To stimulate CeA-CRF neurons selectively, we used optogenetics, in which light-sensitive ion channel-expressing neurons are stimulated using light, and animals underwent several behavioral tasks including operant conditioning for lever pressing, real time place preference task, and elevated plus maze test. Based on the animals’ emotional context (naïve or fear-conditioned), we examined how stimulation of CeA-CRF neurons affected the animals’ behavioral performances. Our results showed that stimulating CeA-CRF in naïve mice produced no effects. However, if the animal’s first experience in a context was appetitive, then stimulating in this context is reinforcing. Similarly, if an animal’s experience is aversive, then stimulating these neurons perpetuates fear and anxiety behaviors. These results suggest that CeA-CRF neurons signal context-dependent behavioral state effects that may be important for engaging appropriate actions in specific contexts.

Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder that is commonly associated with deficits in social, adaptive, and communication skills. Attention-Deficit/Hyperactivity Disorder (ADHD) is characterized by inattention, hyperactivity, and impulsivity that impairs functioning. Previous research estimates that between 30 and 50% of individuals with ASD manifest ADHD symptoms. Although research has shown that individuals with ASD tend to have decreased community involvement, it is not well studied in individuals who have co-occurring ASD and ADHD. The current study explores the relationship between social or community engagement (involvement in organizations, sports, organized group activities) and adaptive skills of individuals with ASD or ASD+ADHD. Participants included 110 youth (m=66, f=44), 8-17 years of age with ASD from the ACE GENDAAR network, a four-site NIH funded project examining sex-based neural differences in children with ASD. All participants included in the sample met ASD criteria on standardized autism assessments (ADOS-2 and ADI-R) and scored ≥70 on a measure of verbal IQ (DAS-II). Parents completed the Child Behavior Checklist (CBCL) reporting on child activity (involvement in sports, organizations, hobbies, chores), ADHD symptoms, overall behavioral problems, and overall competence. Parents also completed the Vineland-II, a parent interview assessing adaptive skills. We hypothesize that there will be a positive correlation between social activity involvement and adaptive skills. That is, children with more community participation will have better adaptive ability. Furthermore, we expect that children with ASD+ADHD compared to children with ASD only, will have greater impairment in adaptive skills and will score lower on the activities scale. The results of this study will provide further understanding of ASD+ADHD and barriers to children participating in community activities and organizations.

Autism spectrum disorder (ASD) is a disorder that is characterized by difficulty in social communication, social skills, and repetitive behavior domains (Sachak, 2016). One of the most prominent features in children with ASD under 3 years of age is delays in language development (Sachak, 2016). This project aims to examine language development in the first year after diagnosis in a sample of preschool children with ASD and to examine family and child demographic characteristics that account for variability in language development. Preschool aged children with ASD (N=59; 7 female) and a matched sample of typically developing (TD) children (N=48; 10 female;) were enrolled in a study of attention and emotion regulation. At enrollment (T1), autism was confirmed using the ADOS module 1 and standardized assessments were done to quantify communication ability (Vineland Adaptive Behavior Scales), nonverbal (visual) reasoning (Mullen Scales of Early Learning), expressive and receptive language (Preschool Language Scale), and self-regulation and executive functioning (Behavior Rating Inventory of Executive Functioning). The PLS, VAB, and BRIEF were repeated at +6 months and +12 months. We hypothesize that: (1) TD children and ASD children who received language interventions will show greater improvement in functional language skills over the first year compared to ASD children ; and (2) children in families with higher household income or education level (one or more parents with college education) will show greater improvements in functional language skills over the 1 year period. Early childhood represents a critical time window for language interventions in order to support functional/adaptive skills and create greater positive outcomes for children with ASD.

Cocaine addiction is a health concern globally. Although both men and women get addicted to cocaine, women transition from casual drug use to addition faster than men, have greater difficulty remaining abstinent and demonstrate greater propensity to relapse. In our laboratory, we found that cocaine craving is potentiated in female rats specifically in the estrus phase of the hormonal cycle. The lateral habenula (LHb), an epithalamic nucleus, plays an important role in cocaine taking and seeking and also mediates several estrogen-dependent behaviors in female rats. In the brain, the effects of estrogen are primarily mediated by estrogen receptors alpha and beta. The goals of this study are - a) to determine if there are hormone-dependent changes in estrogen receptor plasticity in LHb neurons and, b) to define the neuroanatomical organization of estrogen receptors in LHb projection neurons. Firstly, brains from freely-cycling female rats were collected during all phases of the hormonal cycle and analyzed for LHb estrogen receptor expression. Preliminary results indicate that majority of estrogen receptor alpha expressing neurons are located in the ventromedial aspect of the medial to caudal LHb. Analyses are currently underway to determine if cyclical fluctuations in ovarian hormones across the estrus cycle influence estrogen receptor expression in LHb neurons. Secondly, rats were injected with canine adenovirus (CAV2) expressing ZsGreen into either the ventral tegmental area (VTA), dorsal raphe nucleus (DRN) or rostromedial tegmental nucleus (RMTg), three LHb targets. CAV2-ZsGreen vector was retrogradely transported to neuronal cell bodies in the LHb where ZsGreen transgene was expressed. RNAScope in situ hybridization studies are in progress to determine the co-localization of ZsGreen positive neurons in the LHb with estrogen receptors. Understanding the cyclicity of LHb estrogen receptor expression will guide our studies targeted at understanding sex differences in neuronal mechanisms of cocaine taking and seeking behaviors.

Alcohol use disorder (AUD) is associated with severe alcohol-related harm, especially in vulnerable populations. People experiencing homelessness comprise one such population that is disproportionately affected by AUD and its sequelae. Although traditional abstinence-based treatment does not adequately reach or engage this population, a growing body of research has indicated that harm-reduction treatment may present an efficacious alternative. Harm-reduction treatment uses a compassionate and pragmatic approach to help people who use substances and their communities reduce substance-related harm and improve quality of life without requiring abstinence or even use reduction. Primary components of harm-reduction treatment for AUD can include medication-assisted treatment alongside counseling, in which interventionists assess and track harm-reduction metrics, help patients set harm-reduction goals, and discuss safer-use strategies. Studies that have implemented safer-use strategies, or means of staying safer and healthier even if patients are drinking, have shown efficacy in reducing alcohol-related harm. However, no studies to date have explored the associations between implementation of safer-use strategies and alcohol-related harm in people experiencing homelessness and AUD. This secondary study (N=213) was conducted in the context of a larger randomized controlled trial (N=308) of harm-reduction pharmacological and behavioral treatments with people experiencing homelessness and AUD. We aim to quantitatively and qualitatively describe treated patients’ engagement with safer-use strategies over the three-month treatment course and examine the longitudinal association between safer-use strategies and alcohol-related harm. We expect to see inverse associations between the application of safer-use strategies and alcohol-related harm. These findings may help clinicians and counselors better understand what kinds of safer-use strategies are most commonly endorsed by this population and their relative contribution to the reductions in alcohol-related harm and improvements in quality of life observed in the context of harm-reduction treatment with people experiencing homelessness and AUD.

For many years, the primary mode of treatment for people experiencing alcohol use disorder (AUD) has been abstinence-based treatment. Research has indicated, however, that abstinence-based treatment does not optimally engage or treat more severely affected populations, such as people experiencing AUD and homelessness. Instead, harm-reduction treatment approaches are more desirable for this population and can serve as an effective treatment alternative for people experiencing AUD and homelessness. Harm-reduction treatment entails a set of compassionate and pragmatic strategies to emphasize client autonomy, mitigate substance-related harm, and promote quality of life (QoL) without the need for abstinence or use-reduction. Specific components include assessment and tracking of harm-reduction metrics, harm-reduction goal-setting, and implementation of safer-use strategies. This secondary study (N = 213) served to qualitatively and quantitatively explore harm-reduction goals generated by participants in a larger, 4-arm randomized control trial of harm-reduction treatment for people experiencing homelessness and AUD. The three treatment groups included in this secondary study received: a) harm-reduction counseling only, b) harm-reduction counseling + medication assisted treatment (i.e., extended-release naltrexone), and c) harm-reduction counseling + placebo. Participant goals were recorded using the Safer Drinking and Harm Reduction Efforts (SHaRE) scale at baseline assessments and weeks 4, 8, and 12. Qualitative analyses will be conducted to determine the kinds of goals participants generated throughout the 12-week treatment period. Additional descriptive, quantitative analyses will establish quantity of participant goals set at each time point. Finally, inferential statistics will be used to test harm-reduction goals as correlates of alcohol outcomes across the 12-week treatment period. It is expected that a) the combined pharmacotherapy group will generate, progress toward, and achieve more goals than other study conditions; and b) quality-of-life goals will be more strongly associated with reduced alcohol-related harm than drinking-related goals.

An important issue in addressing the cocaine addiction problem is the tendency and probability of recovering addicts to relapse to cocaine after a significant period of time not-using. External cues that have been conditioned to be associated with cocaine use in the past are powerful triggers that cause relapse to cocaine-seeking behaviors. Drug relapse is often caused by experiencing “craving,” a subjective affective state that motivates one to seek drugs. Cocaine craving increasing as a function of time has been shown in the laboratory setting in rats and subsequently dubbed as the “incubation of cocaine craving.” This phenomenon shows that craving increases as a function of time in response to these conditioned cues. Although both women and men have a path to addiction with cocaine, women experience a faster transition from casual use to drug addiction, have a harder time remaining drug-free, and display a greater probability of relapse compared to men. Pre-clinical studies showed that female rats demonstrate an enhanced incubation response when re-exposed to conditioned cocaine cues and display conditioned place preference at lower doses of cocaine than male rats. However, the brain regions that underlie the enhanced incubation response are unknown. We trained cycling female and male rats in the laboratory to self-administer cocaine on a continuous long-access (6h), fixed-ratio 1 reinforcement schedule (reinforcement administered after one correct response) for 10-14 days. After self-administration training, rats were exposed to the cocaine self-administration environment on abstinence days 1 and 35. Our aim for this experiment is to investigate the brain regions that underlie incubation. We are quantifying the immediate early gene c-Fos immunoreactivity in cells across various brain regions in order to determine the similarities and differences in the neural activation during incubation of cocaine craving in female and male rats. We expect to see increased c-Fos expression in animals that are incubuating craving.

Adolescence alcohol use and opioid addiction in adults are systemic issues afflicting the world, and thus, it is important to elucidate the long-term individual and relational consequences of both substance abuse disorders. The purpose of this experiment was to examine the long-term consequences of voluntary adolescent alcohol use on morphine tolerance, fentanyl self-administration, and the effects of previous opioid exposure on fentanyl self-administration in adulthood. Using a preclinical model to examine this hypothesis, adolescent rats had access to alcohol in gelatin form for twenty days, after which a three week withdrawal period occured. Morphine was then administered intraperitoneally for five days and morphine tolerance was measured by a tail-flick test for those five days. Finally, fentanyl self-administration occured in an operant chamber and self-administration will be measure by the amount of fentanyl consumed. My expected results were that adolescent alcohol use will increase morphine tolerance as evidenced by decreased tail-flick time, and fentanyl self-administration will also be increased. Additionally, I expect that previous opioid exposure will increase fentanyl self-administration. Future research should examine the neurobiological mechanisms by which adolescent alcohol use increases morphine tolerance and fentanyl self-administration and how previous opioid exposure increases fentanyl self-administration since these biological mechanism are not well understood.