1 4161 121 MECP2 REGULATES ETHANOL SENSITIVITY AND INTAKE. WE HAVE INVESTIGATED THE EXPRESSION OF CHROMATIN-REGULATING GENES IN THE PREFRONTAL CORTEX AND IN THE SHELL SUBDIVISION OF THE NUCLEUS ACCUMBENS DURING PROTRACTED WITHDRAWAL IN MICE WITH INCREASED ETHANOL DRINKING AFTER CHRONIC INTERMITTENT ETHANOL (CIE) VAPOR EXPOSURE AND IN MICE WITH A HISTORY OF NON-DEPENDENT DRINKING. WE OBSERVED THAT THE METHYL-CPG BINDING PROTEIN 2 (MECP2) WAS ONE OF THE FEW CHROMATIN-REGULATING GENES TO BE DIFFERENTIALLY REGULATED BY A HISTORY OF DEPENDENCE. AS MECP2 HAS THE POTENTIAL OF ACTING AS A BROAD GENE REGULATOR, WE INVESTIGATED SENSITIVITY TO ETHANOL AND ETHANOL DRINKING IN MECP2(308/) (Y) MICE, WHICH HARBOR A TRUNCATED MECP2 ALLELE BUT HAVE A MILDER PHENOTYPE THAN MECP2 NULL MICE. WE OBSERVED THAT MECP2(308/) (Y) MICE WERE MORE SENSITIVE TO ETHANOL'S STIMULATORY AND SEDATIVE EFFECTS THAN WILD-TYPE (WT) MICE, DRANK LESS ETHANOL IN A LIMITED ACCESS 2 BOTTLE CHOICE PARADIGM AND DID NOT SHOW INCREASED DRINKING AFTER INDUCTION OF DEPENDENCE WITH EXPOSURE TO CIE VAPORS. ALCOHOL METABOLISM DID NOT DIFFER IN MECP2(308/) (Y) AND WT MICE. ADDITIONALLY, MECP2(308/) (Y) MICE DID NOT DIFFER FROM WT MICE IN ETHANOL PREFERENCE IN A 24-HOUR PARADIGM NOR IN THEIR INTAKE OF GRADED SOLUTIONS OF SACCHARIN OR QUININE, SUGGESTING THAT THE MECP2(308/) (Y) MUTATION DID NOT ALTER TASTE FUNCTION. LASTLY, USING THE GENE SET ENRICHMENT ANALYSIS ALGORITHM, WE FOUND A SIGNIFICANT OVERLAP IN THE GENES REGULATED BY ALCOHOL AND BY MECP2. TOGETHER, THESE RESULTS SUGGEST THAT MECP2 CONTRIBUTES TO THE REGULATION OF ETHANOL SENSITIVITY AND DRINKING. 2014 2 5855 20 SUBSTANCE USE DISORDER A BIO-DIRECTIONAL SUBSET OF REWARD DEFICIENCY SYNDROME. THIS COMMENTARY IS TO INFORM CLINICIANS CHALLENGED WITH AN INCREASE IN PEOPLE SEEKING TREATMENT FOR SUBSTANCE USE DISORDER (SUD), THAT THE NINETY PERCENT REVOLVING DOOR, IS, IN PART, DUE TO POST-WITHDRAWAL, UNTREATED NEUROTOXICITY. THIS IMPAIRMENT ATTENUATES NEUROTRANSMITTER SIGNALING AND COMPROMISES RESTING STATE FUNCTIONAL CONNECTIVITY, LEADING TO UNWANTED SEQUELAE INCLUDING DEPRESSION, SLEEP DISTURBANCES, SENSATION SEEKING, LACK OF SATISFACTION AND IMPULSIVITY. NEUROIMAGING STUDIES INDICATE THAT NEUROBIOLOGICAL RECOVERY CAN TAKE YEARS. LIKE A "DOUBLE EDGE SWORD" SUD HAS A BIOLOGICAL BI -DIRECTIONAL (BIO-DIRECTIONAL) EFFECT ON THE BRAIN REWARD CIRCUITRY. THE ACUTE INTAKE OF PSYCHOACTIVE DRUGS RESULTS IN HEIGHTENED DOPAMINERGIC ACTIVITY, WHILE, THE OPPOSITE, HYPODOPAMINERGIA OCCURS FOLLOWING CHRONIC ABUSE. INDIVIDUALS WITH SUD CAN HAVE A GENETIC PREDISPOSITION, COMPOUNDED BY STRESS AND NEUROTOXICALLY INDUCED, EPIGENETIC INSULTS THAT IMPACT RECOVERY FROM PROTRACTED ABSTINENCE. FOLLOW-UP POST -SHORT-TERM RECOVERY USUALLY INCLUDES SUPPORTIVE THERAPIES AND PROGRAMS LIKE 12 -STEPS AND OTHER FELLOWSHIPS. HOWEVER, RELAPSE WILL USUALLY OCCUR IF POST -SHORT-TERM RECOVERY HYPODOPAMINERGIA IS NOT TREATED WITH ATTEMPTS AT EPIGENETIC MANIPULATION OF COMPROMISED BRAIN NEUROCHEMISTRY USING SOME MANNER OF PRO-DOPAMINE REGULATION. 2017 3 1610 36 DNA METHYLATION/DEMETHYLATION NETWORK EXPRESSION IN PSYCHOTIC PATIENTS WITH A HISTORY OF ALCOHOL ABUSE. BACKGROUND: RECENT STUDIES SUGGEST THAT PROTRACTED AND EXCESSIVE ALCOHOL USE INDUCES AN EPIGENETIC DYSREGULATION IN HUMAN AND RODENT BRAINS. WE RECENTLY REPORTED THAT DNA METHYLATION DYNAMICS ARE ALTERED IN BRAINS OF PSYCHOTIC (PS) PATIENTS, INCLUDING SCHIZOPHRENIA AND BIPOLAR DISORDER PATIENTS. BECAUSE PS PATIENTS ARE OFTEN COMORBID WITH CHRONIC ALCOHOL ABUSE, WE EXAMINED WHETHER THE ALTERED EXPRESSION OF MULTIPLE MEMBERS OF THE DNA METHYLATION/DEMETHYLATION NETWORK OBSERVED IN POSTMORTEM BRAINS OF PS PATIENTS WAS MODIFIED IN PS PATIENTS WITH A HISTORY OF CHRONIC ALCOHOL ABUSE. METHODS: DNA-METHYLTRANSFERASE-1 (DNMT1) MRNA-POSITIVE NEURONS WERE COUNTED IN SITU IN PREFRONTAL CORTEX SAMPLES OBTAINED FROM THE HARVARD BRAIN TISSUE RESOURCE CENTER, BELMONT, MA. 10-11-TRANSLOCATION (TETS 1, 2, 3), APOLIPOPROTEIN B EDITING COMPLEX ENZYME (APOBEC-3C), GROWTH AND DNA-DAMAGE-INDUCIBLE PROTEIN 45BETA (GADD45BETA), AND METHYL-BINDING DOMAIN PROTEIN-4 (MBD4) MRNAS WERE MEASURED BY QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION IN INFERIOR PARIETAL CORTICAL LOBULE SAMPLES OBTAINED FROM THE STANLEY FOUNDATION NEUROPATHOLOGY CONSORTIUM, BETHESDA, MD. RESULTS: WE OBSERVED AN INCREASE IN DNMT1 MRNA-POSITIVE NEURONS IN PS PATIENTS COMPARED WITH NON-PS SUBJECTS. IN ADDITION, THERE WAS A PRONOUNCED DECREASE IN APOBEC-3C AND A PRONOUNCED INCREASE IN GADD45BETA AND TET1 MRNAS IN PS PATIENTS WITH NO HISTORY OF ALCOHOL ABUSE. IN PS PATIENTS WITH A HISTORY OF CHRONIC ALCOHOL ABUSE, THE NUMBERS OF DNMT1-POSITIVE NEURONS WERE NOT INCREASED SIGNIFICANTLY. FURTHERMORE, THE DECREASE IN APOBEC-3C MRNA WAS LESS PRONOUNCED, WHILE THE INCREASE IN TET1 MRNA HAD A TENDENCY TO BE POTENTIATED IN THOSE PS PATIENTS THAT WERE CHRONIC ALCOHOL ABUSERS. GADD45BETA AND MBD4 MRNAS WERE NOT INFLUENCED BY ALCOHOL ABUSE. THE EFFECT OF CHRONIC ALCOHOL ABUSE ON DNA METHYLATION/DEMETHYLATION NETWORK ENZYMES CANNOT BE ATTRIBUTED TO CONFOUNDING DEMOGRAPHIC VARIABLES OR TO THE TYPE AND DOSE OF MEDICATION USED. CONCLUSIONS: BASED ON THESE RESULTS, WE HYPOTHESIZE THAT PS PATIENTS MAY ABUSE ALCOHOL AS A POTENTIAL ATTEMPT AT SELF-MEDICATION TO NORMALIZE ALTERED DNA METHYLATION/DEMETHYLATION NETWORK PATHWAYS. HOWEVER, BEFORE ACCEPTING THIS CONCLUSION, WE NEED TO STUDY ALTERATIONS IN THE DNA METHYLATION/DEMETHYLATION PATHWAYS AND THE DNA METHYLATION DYNAMICS IN A SUBSTANTIAL NUMBER OF ALCOHOLIC PS AND NON-PS PATIENTS. ADDITIONAL INVESTIGATION MAY ALSO BE NECESSARY TO DETERMINE WHETHER THE ALTERED DNA METHYLATION DYNAMICS ARE DIRECT OR THE CONSEQUENCE OF AN INDIRECT INTERACTION OF ALCOHOL WITH THE NEUROPATHOGENETIC MECHANISMS UNDERLYING PSYCHOSIS. 2013 4 5791 40 STABLE HISTONE METHYLATION CHANGES AT PROTEOGLYCAN NETWORK GENES FOLLOWING ETHANOL EXPOSURE. ALCOHOL USE DISORDER (AUD) IS A CHRONIC MENTAL ILLNESS IN WHICH PATIENTS OFTEN ACHIEVE PROTRACTED PERIODS OF ABSTINENCE PRIOR TO RELAPSE. EPIGENETIC MECHANISMS MAY PROVIDE AN EXPLANATION FOR THE PERSISTING GENE EXPRESSION CHANGES THAT CAN BE OBSERVED EVEN AFTER LONG PERIODS OF ABSTINENCE AND MAY CONTRIBUTE TO RELAPSE. IN THIS STUDY, WE EXAMINED TWO HISTONE MODIFICATIONS, HISTONE 3 LYSINE 4 TRI-METHYLATION (H3K4ME3) AND HISTONE 3 LYSINE 27 TRI-METHYLATION (H3K27ME3), IN THE PREFRONTAL CORTEX OF WITHDRAWAL SEIZURE RESISTANT (WSR) MICE 21 DAYS AFTER 72 H OF ETHANOL VAPOR EXPOSURE. THESE HISTONE MODIFICATIONS WERE SELECTED BECAUSE THEY ARE ASSOCIATED WITH ACTIVE PROMOTERS (H3K4ME3) AND REPRESSED GENE EXPRESSION IN A EUCHROMATIC ENVIRONMENT (H3K27ME3). WE PERFORMED A GENOME-WIDE ANALYSIS TO IDENTIFY DIFFERENCES IN H3K4ME3 AND H3K27ME3 LEVELS IN POST-ETHANOL EXPOSURE VS. CONTROL MICE BY CHIP-SEQ. WE DETECTED A GLOBAL REDUCTION IN H3K4ME3 PEAKS AND INCREASE IN H3K27ME3 PEAKS IN POST-ETHANOL EXPOSURE MICE COMPARED TO CONTROLS, THESE CHANGES ARE CONSISTENT WITH PERSISTENT REDUCTIONS IN GENE EXPRESSION. PATHWAY ANALYSIS OF GENES DISPLAYING CHANGES IN H3K4ME3 AND H3K27ME3 REVEALED ENRICHMENT FOR GENES INVOLVED IN PROTEOGLYCAN AND CALCIUM SIGNALING PATHWAYS, RESPECTIVELY. MICROARRAY ANALYSIS OF 7,683 GENES AND QPCR ANALYSIS IDENTIFIED EIGHT GENES DISPLAYING CONCORDANT REGULATION OF GENE EXPRESSION AND H3K4ME3/H3K27ME3. WE ALSO COMPARED CHANGES IN H3K4ME3 AND/OR H3K27ME3 FROM OUR STUDY WITH CHANGES IN GENE EXPRESSION IN RESPONSE TO ETHANOL FROM PUBLISHED LITERATURE AND WE FOUND THAT THE EXPRESSION OF 52% OF THE GENES WITH ALTERED H3K4ME3 BINDING AND 40% OF GENES WITH H3K27ME3 DIFFERENCES ARE ALTERED BY ETHANOL EXPOSURE. THE CHROMATIN CHANGES ASSOCIATED WITH THE 21-DAY POST-EXPOSURE PERIOD SUGGEST THAT THIS PERIOD IS A UNIQUE STATE IN THE ADDICTION CYCLE THAT DIFFERS FROM ETHANOL INTOXICATION AND ACUTE WITHDRAWAL. THESE RESULTS PROVIDE INSIGHTS INTO THE ENDURING EFFECTS OF ETHANOL ON PROTEOGLYCAN AND CALCIUM SIGNALING GENES IN THE BRAIN. 2018 5 1731 40 DYSREGULATION OF THE HISTONE DEMETHYLASE KDM6B IN ALCOHOL DEPENDENCE IS ASSOCIATED WITH EPIGENETIC REGULATION OF INFLAMMATORY SIGNALING PATHWAYS. EPIGENETIC ENZYMES OVERSEE LONG-TERM CHANGES IN GENE EXPRESSION BY INTEGRATING GENETIC AND ENVIRONMENTAL CUES. WHILE THERE ARE HUNDREDS OF ENZYMES THAT CONTROL HISTONE AND DNA MODIFICATIONS, THEIR POTENTIAL ROLES IN SUBSTANCE ABUSE AND ALCOHOL DEPENDENCE REMAIN UNDEREXPLORED. A FEW RECENT STUDIES HAVE SUGGESTED THAT EPIGENETIC PROCESSES COULD UNDERLIE TRANSCRIPTOMIC AND BEHAVIORAL HALLMARKS OF ALCOHOL ADDICTION. IN THE PRESENT STUDY, WE SOUGHT TO IDENTIFY EPIGENETIC ENZYMES IN THE BRAIN THAT ARE DYSREGULATED DURING PROTRACTED ABSTINENCE AS A CONSEQUENCE OF CHRONIC AND INTERMITTENT ALCOHOL EXPOSURE. THROUGH QUANTITATIVE MRNA EXPRESSION ANALYSIS OF OVER 100 EPIGENETIC ENZYMES, WE IDENTIFIED 11 THAT ARE SIGNIFICANTLY ALTERED IN ALCOHOL-DEPENDENT RATS COMPARED WITH CONTROLS. FOLLOW-UP STUDIES OF ONE OF THESE ENZYMES, THE HISTONE DEMETHYLASE KDM6B, SHOWED THAT THIS ENZYME EXHIBITS REGION-SPECIFIC DYSREGULATION IN THE PREFRONTAL CORTEX AND NUCLEUS ACCUMBENS OF ALCOHOL-DEPENDENT RATS. KDM6B WAS ALSO UPREGULATED IN THE HUMAN ALCOHOLIC BRAIN. UPREGULATION OF KDM6B PROTEIN IN ALCOHOL-DEPENDENT RATS WAS ACCOMPANIED BY A DECREASE OF TRIMETHYLATION LEVELS AT HISTONE H3, LYSINE 27 (H3K27ME3), CONSISTENT WITH THE KNOWN DEMETHYLASE SPECIFICITY OF KDM6B. SUBSEQUENT EPIGENETIC (CHROMATIN IMMUNOPRECIPITATION [CHIP]-SEQUENCING) ANALYSIS SHOWED THAT ALCOHOL-INDUCED CHANGES IN H3K27ME3 WERE SIGNIFICANTLY ENRICHED AT GENES IN THE IL-6 SIGNALING PATHWAY, CONSISTENT WITH THE WELL-CHARACTERIZED ROLE OF KDM6B IN MODULATION OF INFLAMMATORY RESPONSES. KNOCKDOWN OF KDM6B IN CULTURED MICROGLIAL CELLS DIMINISHED IL-6 INDUCTION IN RESPONSE TO AN INFLAMMATORY STIMULUS. OUR FINDINGS IMPLICATE A NOVEL KDM6B-MEDIATED EPIGENETIC SIGNALING PATHWAY INTEGRATED WITH INFLAMMATORY SIGNALING PATHWAYS THAT ARE KNOWN TO UNDERLIE THE DEVELOPMENT OF ALCOHOL ADDICTION. 2021 6 5342 25 RADIATION-INDUCED LATE EFFECTS IN TWO AFFECTED INDIVIDUALS OF THE LILO RADIATION ACCIDENT. RADIATION EXPOSURE LEADS TO A RISK FOR LONG-TERM DETERMINISTIC AND STOCHASTIC LATE EFFECTS. TWO INDIVIDUALS EXPOSED TO PROTRACTED PHOTON RADIATION IN THE RADIOLOGICAL ACCIDENT AT THE LILO MILITARY SITE IN GEORGIA IN 1997 RECEIVED FOLLOW-UP TREATMENT AND RESECTION OF SEVERAL CHRONIC RADIATION ULCERS IN THE BUNDESWEHR HOSPITAL ULM, GERMANY, IN 2003. MULTI-PARAMETER ANALYSIS REVEALED THAT SPERMATOGENETIC ARREST AND SERUM HORMONE LEVELS IN BOTH PATIENTS HAD RECOVERED COMPARED TO THE STATUS IN 1997. HOWEVER, WE OBSERVED A PERSISTENCE OF ALTERED T-CELL RATIOS, INCREASED ICAM1 AND BETA1-INTEGRIN EXPRESSION, AND ABERRANT BONE MARROW CELLS AND LYMPHOCYTES WITH SIGNIFICANTLY INCREASED TRANSLOCATIONS 6 YEARS AFTER THE ACCIDENT. THIS INVESTIGATION THUS IDENTIFIED ALTERED END POINTS STILL DETECTABLE YEARS AFTER THE ACCIDENT THAT SUGGEST PERSISTENT GENOMIC DAMAGE AS WELL AS EPIGENETIC EFFECTS IN THESE INDIVIDUALS, WHICH MAY BE ASSOCIATED WITH AN ELEVATED RISK FOR THE DEVELOPMENT OF FURTHER LATE EFFECTS. OUR OBSERVATIONS FURTHER SUGGEST THE DEVELOPMENT OF A CHRONIC RADIATION SYNDROME AND INDICATE FOLLOW-UP PARAMETERS IN RADIATION VICTIMS. 2007 7 1850 26 ELECTROACUPUNCTURE AMELIORATES DEPRESSION-LIKE BEHAVIORS COMORBID TO CHRONIC NEUROPATHIC PAIN VIA TET1-MEDIATED RESTORATION OF ADULT NEUROGENESIS. ALTHOUGH ELECTROACUPUNCTURE (EA) STIMULATION IS A WIDELY USED THERAPY FOR CHRONIC PAIN AND COMORBID PSYCHIATRIC DISORDERS, ITS LONG-TERM EFFECTS ON CHRONIC NEUROPATHIC PAIN-INDUCED DEPRESSION AND THE UNDERLYING MECHANISMS REMAIN ELUSIVE. IN THE PRESENT STUDY, WE FOUND THAT EA STIMULATION WAS ABLE TO RESTORE ADULT NEUROGENESIS IN THE VENTRAL DENTATE GYRUS (DG), BY BOTH INCREASING NEURONAL DIFFERENTIATION AND RESTORING THE NORMAL MORPHOLOGY OF NEWBORN DENDRITES, IN MICE WITH SPARED NERVE INJURY SURGERY. BY ABLATING THE NESTIN+ NEURAL STEM CELLS (NSCS) VIA DIPHTHERIA TOXIN FRAGMENT A EXPRESSION, WE FURTHER PROVED THAT NEUROGENESIS IN THE VENTRAL DG WAS CRUCIAL TO THE LONG-TERM, BUT NOT THE IMMEDIATE ANTIDEPRESSANT EFFECT OF EA, NOR WAS IT ASSOCIATED WITH NOCICEPTION. FURTHERMORE, WE FOUND THAT THE RESTORATION OF NEUROGENESIS WAS DEPENDENT ON TET1-MEDIATED EPIGENETIC MODIFICATION UPON EA TREATMENT. TET1 COULD BIND TO THE PROMOTER OF THE PROX1 GENE, THUS CATALYZING ITS DEMETHYLATION AND FACILITATING ITS EXPRESSION, WHICH FINALLY CONTRIBUTED TO THE RESTORATION OF NEUROGENESIS AND AMELIORATION OF DEPRESSION-LIKE BEHAVIORS INDUCED BY CHRONIC NEUROPATHIC PAIN. THUS, WE CONCLUDE THAT EA STIMULATION RESTORES INHIBITED TET1 EXPRESSION IN HIPPOCAMPAL NSCS OF MICE WITH CHRONIC NEUROPATHIC PAIN, AND INCREASED TET1 EXPRESSION AMELIORATES HYPERMETHYLATION OF PROX1 AND RESTORES NORMAL ADULT NEUROGENESIS IN THE VENTRAL DG, WHICH CONTRIBUTES TO THE LONG-TERM ANTIDEPRESSANT EFFECT OF EA. 2023 8 6177 35 THE HISTONE METHYLTRANSFERASE G9A MEDIATES STRESS-REGULATED ALCOHOL DRINKING. THE EPIGENETIC ENZYME G9A IS A HISTONE METHYLTRANSFERASE THAT DIMETHYLATES LYSINE 9 ON HISTONE H3 (H3K9ME2), AND IN THE ADULT NUCLEUS ACCUMBENS (NAC), G9A REGULATES MULTIPLE BEHAVIORS ASSOCIATED WITH SUBSTANCE USE DISORDER. WE SHOW HERE THAT CHRONIC INTERMITTENT ETHANOL (CIE) EXPOSURE IN MALE MICE REDUCED BOTH G9A AND H3K9ME2 LEVELS IN THE ADULT NAC, BUT NOT DORSAL STRIATUM. VIRAL-MEDIATED REDUCTION OF G9A IN THE NAC HAD NO EFFECTS ON BASELINE VOLITIONAL ETHANOL DRINKING OR ESCALATED ALCOHOL DRINKING PRODUCED BY CIE EXPOSURE; HOWEVER, NAC G9A WAS REQUIRED FOR STRESS-REGULATED CHANGES IN ETHANOL DRINKING, INCLUDING POTENTIATED ALCOHOL DRINKING PRODUCED BY ACTIVATION OF THE KAPPA-OPIOID RECEPTOR. IN ADDITION, WE OBSERVED THAT CHRONIC SYSTEMIC ADMINISTRATION OF A G9A INHIBITOR, UNC0642, ALSO BLOCKED STRESS-POTENTIATED ALCOHOL DRINKING. TOGETHER, OUR FINDINGS SUGGEST THAT CHRONIC ALCOHOL USE, SIMILAR TO OTHER ABUSED SUBSTANCES, PRODUCES A NAC-SELECTIVE REDUCTION IN G9A LEVELS THAT SERVES TO LIMIT STRESS-REGULATED ALCOHOL DRINKING. MOREOVER, OUR FINDINGS SUGGEST THAT PHARMACOLOGICAL INHIBITION OF G9A MIGHT PROVIDE A NOVEL THERAPEUTIC APPROACH TO TREAT STRESS-INDUCED ALCOHOL DRINKING, WHICH IS A MAJOR TRIGGER OF RELAPSE IN INDIVIDUALS SUFFERING FROM AUD. 2022 9 5015 27 PERSISTENCE OF SELF-REACTIVE CD8+ T CELLS IN THE CNS REQUIRES TOX-DEPENDENT CHROMATIN REMODELING. SELF-REACTIVE CD8(+) T CELLS ARE IMPORTANT MEDIATORS OF PROGRESSIVE TISSUE DAMAGE IN AUTOIMMUNE DISEASES, BUT THE MOLECULAR PROGRAM UNDERLYING THESE CELLS' FUNCTIONAL ADAPTATION IS UNCLEAR. HERE WE CHARACTERIZE THE TRANSCRIPTIONAL AND EPIGENETIC LANDSCAPE OF SELF-REACTIVE CD8(+) T CELLS IN A MOUSE MODEL OF PROTRACTED CENTRAL NERVOUS SYSTEM (CNS) AUTOIMMUNITY AND COMPARE IT TO POPULATIONS OF CNS-RESIDENT MEMORY CD8(+) T CELLS EMERGING FROM ACUTE VIRAL INFECTION. WE FIND THAT AUTOIMMUNE CD8(+) T CELLS PERSISTING AT SITES OF SELF-ANTIGEN EXHIBIT CHARACTERISTIC TRANSCRIPTIONAL REGULATION TOGETHER WITH DISTINCT EPIGENETIC REMODELING. THIS SELF-REACTIVE CD8(+) T CELL FATE DEPENDS ON THE TRANSCRIPTIONAL REGULATION BY THE DNA-BINDING HMG-BOX PROTEIN TOX WHICH REMODELS MORE THAN 400 GENOMIC REGIONS INCLUDING LOCI SUCH AS TCF7, WHICH IS CENTRAL TO STEMNESS OF CD8(+) T CELLS. CONTINUOUS EXPOSURE TO CNS SELF-ANTIGEN SUSTAINS TOX LEVELS IN SELF-REACTIVE CD8(+) T CELLS, WHEREAS GENETIC ABLATION OF TOX IN CD8(+) T CELLS RESULTS IN SHORTENED PERSISTENCE OF SELF-REACTIVE CD8(+) T CELLS IN THE INFLAMED CNS. OUR STUDY ESTABLISHES AND CHARACTERIZES THE GENETIC DIFFERENTIATION PROGRAM ENABLING CHRONIC T CELL-DRIVEN IMMUNOPATHOLOGY IN CNS AUTOIMMUNITY. 2021 10 5177 47 PREFRONTAL CORTEX EXPRESSION OF CHROMATIN MODIFIER GENES IN MALE WSP AND WSR MICE CHANGES ACROSS ETHANOL DEPENDENCE, WITHDRAWAL, AND ABSTINENCE. ALCOHOL-USE DISORDER (AUD) IS A RELAPSING DISORDER ASSOCIATED WITH EXCESSIVE ETHANOL CONSUMPTION. RECENT STUDIES SUPPORT THE INVOLVEMENT OF EPIGENETIC MECHANISMS IN THE DEVELOPMENT OF AUD. STUDIES CARRIED OUT SO FAR HAVE FOCUSED ON A FEW SPECIFIC EPIGENETIC MODIFICATIONS. THE GOAL OF THIS PROJECT WAS TO INVESTIGATE GENE EXPRESSION CHANGES OF EPIGENETIC REGULATORS THAT MEDIATE A BROAD ARRAY OF CHROMATIN MODIFICATIONS AFTER CHRONIC ALCOHOL EXPOSURE, CHRONIC ALCOHOL EXPOSURE FOLLOWED BY 8 H WITHDRAWAL, AND CHRONIC ALCOHOL EXPOSURE FOLLOWED BY 21 DAYS OF ABSTINENCE IN WITHDRAWAL-RESISTANT (WSR) AND WITHDRAWAL SEIZURE-PRONE (WSP) SELECTED MOUSE LINES. WE FOUND THAT CHRONIC VAPOR EXPOSURE TO HIGHLY INTOXICATING LEVELS OF ETHANOL ALTERS THE EXPRESSION OF SEVERAL CHROMATIN REMODELING GENES MEASURED BY QUANTITATIVE PCR ARRAY ANALYSES. THE IDENTIFIED EFFECTS WERE INDEPENDENT OF SELECTED LINES, WHICH, HOWEVER, DISPLAYED BASELINE DIFFERENCES IN EPIGENETIC GENE EXPRESSION. WE REPORTED DYSREGULATION IN THE EXPRESSION OF GENES INVOLVED IN HISTONE ACETYLATION, DEACETYLATION, LYSINE AND ARGININE METHYLATION AND UBIQUITINATIONHYLATION DURING CHRONIC ETHANOL EXPOSURE AND WITHDRAWAL, BUT NOT AFTER 21 DAYS OF ABSTINENCE. ETHANOL-INDUCED CHANGES ARE CONSISTENT WITH DECREASED HISTONE ACETYLATION AND WITH DECREASED DEPOSITION OF THE PERMISSIVE UBIQUITINATION MARK H2BK120UB, ASSOCIATED WITH REDUCED TRANSCRIPTION. ON THE OTHER HAND, ETHANOL-INDUCED CHANGES IN THE EXPRESSION OF GENES INVOLVED IN HISTONE LYSINE METHYLATION ARE CONSISTENT WITH INCREASED TRANSCRIPTION. THE NET RESULT OF THESE MODIFICATIONS ON GENE EXPRESSION IS LIKELY TO DEPEND ON THE COMBINATION OF THE SPECIFIC HISTONE TAIL MODIFICATIONS PRESENT AT A GIVEN TIME ON A GIVEN PROMOTER. SINCE ALCOHOL DOES NOT MODULATE GENE EXPRESSION UNIDIRECTIONALLY, IT IS NOT SURPRISING THAT ALCOHOL DOES NOT UNIDIRECTIONALLY ALTER CHROMATIN STRUCTURE TOWARD A CLOSED OR OPEN STATE, AS SUGGESTED BY THE RESULTS OF THIS STUDY. 2017 11 4943 33 PATERNAL PRECONCEPTION ALCOHOL EXPOSURE IMPARTS INTERGENERATIONAL ALCOHOL-RELATED BEHAVIORS TO MALE OFFSPRING ON A PURE C57BL/6J BACKGROUND. WHILE ALCOHOL USE DISORDER (AUD) IS A HIGHLY HERITABLE CONDITION, THE BASIS OF AUD IN FAMILIES WITH A HISTORY OF ALCOHOLISM IS DIFFICULT TO EXPLAIN BY GENETIC VARIATION ALONE. EMERGING EVIDENCE SUGGESTS THAT PARENTAL EXPERIENCE PRIOR TO CONCEPTION CAN AFFECT INHERITANCE OF COMPLEX BEHAVIORS IN OFFSPRING VIA NON-GENOMIC (EPIGENETIC) MECHANISMS. FOR INSTANCE, MALE C57BL/6J (B6) MICE EXPOSED TO CHRONIC INTERMITTENT VAPOR ETHANOL (CIE) PRIOR TO MATING WITH STRAIN 129S1/SVIMJ ETHANOL-NAIVE FEMALES PRODUCE MALE OFFSPRING WITH REDUCED ETHANOL-DRINKING PREFERENCE, INCREASED ETHANOL SENSITIVITY, AND INCREASED BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) EXPRESSION IN THE VENTRAL TEGMENTAL AREA (VTA). IN THE PRESENT STUDY, WE TESTED THE HYPOTHESIS THAT THESE INTERGENERATIONAL EFFECTS OF PATERNAL CIE ARE REPRODUCIBLE IN MALE OFFSPRING ON AN INBRED B6 BACKGROUND. TO THIS END, B6 MALES WERE EXPOSED TO 6 WEEKS OF CIE (OR ROOM AIR AS A CONTROL) BEFORE MATING WITH ETHANOL-NAIVE B6 FEMALES TO PRODUCE ETHANOL (E)-SIRED AND CONTROL (C)-SIRED MALE AND FEMALE OFFSPRING. WE OBSERVED A SEX-SPECIFIC EFFECT, AS E-SIRED MALES EXHIBITED DECREASED TWO-BOTTLE FREE-CHOICE ETHANOL-DRINKING PREFERENCE, INCREASED SENSITIVITY TO THE ANXIOLYTIC EFFECTS OF ETHANOL, AND INCREASED VTA BDNF EXPRESSION; NO DIFFERENCES WERE OBSERVED IN FEMALE OFFSPRING. THESE FINDINGS CONFIRM AND EXTEND OUR PREVIOUS RESULTS BY DEMONSTRATING THAT THE EFFECTS OF PATERNAL PRECONCEPTION ETHANOL ARE REPRODUCIBLE USING GENETICALLY IDENTICAL, INBRED B6 ANIMALS. 2017 12 2770 27 EXTENDING INJURY- AND DISEASE-RESISTANT CNS PHENOTYPES BY REPETITIVE EPIGENETIC CONDITIONING. SIGNIFICANT REDUCTIONS IN THE EXTENT OF ACUTE INJURY IN THE CNS CAN BE ACHIEVED BY EXPOSURE TO DIFFERENT PRECONDITIONING STIMULI, BUT THE DURATION OF THE INDUCED PROTECTIVE PHENOTYPE IS TYPICALLY SHORT-LASTING, AND THUS IS DEEMED AS LIMITING ITS CLINICAL APPLICABILITY. EXTENDING THE PERIOD OVER WHICH SUCH ADAPTIVE EPIGENETIC CHANGES PERSIST - IN EFFECT, EXPANDING CONDITIONING'S "THERAPEUTIC WINDOW" - WOULD SIGNIFICANTLY BROADEN THE POTENTIAL APPLICATIONS OF SUCH A TREATMENT APPROACH IN PATIENTS. THE FREQUENCY OF THE CONDITIONING STIMULUS MAY HOLD THE KEY. WHILE TRANSIENT (1-3 DAYS) PROTECTION AGAINST CNS ISCHEMIC INJURY IS WELL ESTABLISHED PRECLINICALLY FOLLOWING A SINGLE PRECONDITIONING STIMULUS, REPETITIVELY PRESENTING PRECONDITIONING STIMULI EXTENDS THE DURATION OF ISCHEMIC TOLERANCE BY MANY WEEKS. MOREOVER, REPETITIVE INTERMITTENT POSTCONDITIONING ENHANCES POST-ISCHEMIC RECOVERY METRICS AND IMPROVES LONG-TERM SURVIVAL. INTERMITTENT CONDITIONING IS ALSO EFFICACIOUS FOR PREVENTING OR DELAYING INJURY IN PRECLINICAL MODELS OF CHRONIC NEURODEGENERATIVE DISEASE, AND FOR PROMOTING LONG-LASTING FUNCTIONAL IMPROVEMENTS IN A NUMBER OF OTHER PATHOLOGIES AS WELL. ALTHOUGH THE DETAILED MECHANISMS UNDERLYING THESE PROTRACTED KINDS OF NEUROPLASTICITY REMAIN LARGELY UNSTUDIED, ACCUMULATING EMPIRICAL EVIDENCE SUPPORTS THE CONTENTION THAT ALL OF THESE ADAPTIVE PHENOTYPES ARE EPIGENETICALLY MEDIATED. GOING FORWARD, ADDITIONAL PRECLINICAL DEMONSTRATIONS OF THE ABILITY TO INDUCE SUSTAINED BENEFICIAL PHENOTYPES THAT REDUCE THE BURDEN OF ACUTE AND CHRONIC NEURODEGENERATION, AND EXPERIMENTAL INTERROGATIONS OF THE REGULATORY CONSTRUCTS RESPONSIBLE FOR THESE EPIGENETIC RESPONSES, WILL ACCELERATE THE IDENTIFICATION OF NOT ONLY EFFICACIOUS BUT ALSO PRACTICAL, ADAPTIVE EPIGENETICS-BASED TREATMENTS FOR INDIVIDUALS WITH NEUROLOGICAL DISEASE. 2015 13 5445 44 REPEATED VAPOR ETHANOL EXPOSURE INDUCES TRANSIENT HISTONE MODIFICATIONS IN THE BRAIN THAT ARE MODIFIED BY GENOTYPE AND BRAIN REGION. BACKGROUND: EMERGING RESEARCH IMPLICATES ETHANOL (ETOH)-INDUCED EPIGENETIC MODIFICATIONS IN REGULATING GENE EXPRESSION AND ETOH CONSUMPTION. HOWEVER, CONSENSUS ON SPECIFIC EPIGENETIC MODIFICATIONS INDUCED BY ETOH HAS NOT YET EMERGED, MAKING IT CHALLENGING TO IDENTIFY MECHANISMS AND DEVELOP TARGETED TREATMENTS. WE HYPOTHESIZED THAT CHRONIC INTERMITTENT ETOH (CIE) INDUCES PERSISTENT CHANGES IN HISTONE MODIFICATIONS ACROSS THE CEREBRAL CORTEX (CCX), NUCLEUS ACCUMBENS (NAC), AND PREFRONTAL CORTEX (PFC), AND THAT THESE HISTONE MODIFICATIONS ARE ALTERED IN A KNOCK-IN MOUSE STRAIN WITH ALTERED SENSITIVITY TO ETOH. METHODS: C57BL/6J (B6) MICE AND ALPHA1SHLA KNOCKIN MICE ON A B6 BACKGROUND WERE EXPOSED TO 16 H OF VAPOR ETOH OR ROOM AIR FOLLOWED BY 8 H OF ROOM AIR FOR 4 CONSECUTIVE DAYS AND SACRIFICED AT MULTIPLE TIME POINTS UP TO 72 H FOLLOWING EXPOSURE. HISTONE MODIFICATIONS WERE ASSESSED USING WESTERN BLOT AND DOT BLOT. RT-QPCR WAS USED TO STUDY EXPRESSION OF CHROMATIN MODIFYING ENZYMES IN NAC AND PFC. RESULTS: IN NAC, CIE SIGNIFICANTLY INCREASED ACETYLATION OF HISTONE SUBUNIT H3 AT LYSINE 9 (H3K9AC) BUT NOT LYSINE 14 (H3K14AC) OR LYSINE 27 (H3K27AC). IN PFC, CIE SIGNIFICANTLY INCREASED H3K9AC BUT NOT H3K14 OR H3K27AC. THERE WERE NO SIGNIFICANT CHANGES AT 8 OR 72 H AFTER ETOH EXPOSURE IN EITHER NAC OR PFC. CIE WAS ALSO ASSOCIATED WITH INCREASED EXPRESSION OF KAT2B, KAT5, AND TET1 IN NAC BUT NOT PFC. IN CCX, CIE HAD A SIGNIFICANT EFFECT ON LEVELS OF H3K18AC; THERE WAS ALSO A SIGNIFICANT EFFECT OF THE ALPHA1SHLA MUTATION ON LEVELS OF H3K27ME3, H3K14AC, AND H3K18AC AS WELL AS A TREND FOR H3S10PK14AC. CONCLUSIONS: THE ETOH-INDUCED HISTONE MODIFICATIONS OBSERVED WERE TRANSIENT AND VARIED SIGNIFICANTLY BETWEEN BRAIN REGIONS. A GENETIC MUTATION THAT ALTERED SENSITIVITY TO ETOH WAS ASSOCIATED WITH ALTERED INDUCTION OF HISTONE MODIFICATIONS DURING CIE. THESE RESULTS HAVE IMPLICATIONS FOR STUDYING ETOH-INDUCED HISTONE MODIFICATIONS AND ETOH SENSITIVITY. 2015 14 4878 37 OVEREXPRESSION OF THE HISTONE DIMETHYLTRANSFERASE G9A IN NUCLEUS ACCUMBENS SHELL INCREASES COCAINE SELF-ADMINISTRATION, STRESS-INDUCED REINSTATEMENT, AND ANXIETY. REPEATED EXPOSURE TO COCAINE INDUCES LASTING EPIGENETIC CHANGES IN NEURONS THAT PROMOTE THE DEVELOPMENT AND PERSISTENCE OF ADDICTION. ONE EPIGENETIC ALTERATION INVOLVES REDUCTIONS IN LEVELS OF THE HISTONE DIMETHYLTRANSFERASE G9A IN NUCLEUS ACCUMBENS (NAC) AFTER CHRONIC COCAINE ADMINISTRATION. THIS REDUCTION IN G9A MAY ENHANCE COCAINE REWARD BECAUSE OVEREXPRESSING G9A IN THE NAC DECREASES COCAINE-CONDITIONED PLACE PREFERENCE. THEREFORE, WE HYPOTHESIZED THAT HSV-MEDIATED G9A OVEREXPRESSION IN THE NAC SHELL (NACSH) WOULD ATTENUATE COCAINE SELF-ADMINISTRATION (SA) AND COCAINE-SEEKING BEHAVIOR. INSTEAD, WE FOUND THAT G9A OVEREXPRESSION, AND THE RESULTING INCREASE IN HISTONE 3 LYSINE 9 DIMETHYLATION (H3K9ME2), INCREASES SENSITIVITY TO COCAINE REINFORCEMENT AND ENHANCES MOTIVATION FOR COCAINE IN SELF-ADMINISTERING MALE RATS. MOREOVER, WHEN G9A OVEREXPRESSION IS LIMITED TO THE INITIAL 15 D OF COCAINE SA TRAINING, IT PRODUCES AN ENDURING POSTEXPRESSION ENHANCEMENT IN COCAINE SA AND PROLONGED (OVER 5 WEEKS) INCREASES IN REINSTATEMENT OF COCAINE SEEKING INDUCED BY FOOT-SHOCK STRESS, BUT IN THE ABSENCE OF CONTINUED GLOBAL ELEVATIONS IN H3K9ME2. THE INCREASE IN STRESS-INDUCED REINSTATEMENT IS PARALLELED BY HEIGHTENED ANXIETY MEASURES, SUGGESTING THAT COUNTERING THE COCAINE-INDUCED DECREASES IN ENDOGENOUS G9A WITH ECTOPIC G9A OVEREXPRESSION LEADS TO LASTING ANXIOGENIC EFFECTS. FINALLY, WE FOUND AN ENDURING REDUCTION IN PHOSPHORYLATED CAMP-RESPONSE ELEMENT BINDING PROTEIN LEVELS IN THE NACSH THAT COULD ACCOUNT FOR THE INCREASED ANXIETY. THESE DATA DEMONSTRATE A NOVEL ROLE FOR G9A IN PROMOTING COMORBID COCAINE ADDICTION AND ANXIETY AND SUGGEST THAT INCREASED EPIGENETIC REPRESSION OF TRANSCRIPTION THROUGH H3K9 DURING COCAINE USE CAN HAVE LONG-LASTING AND UNEXPECTED NEGATIVE CONSEQUENCES ON BEHAVIOR.SIGNIFICANCE STATEMENT COCAINE ADDICTION IS A NEUROPSYCHIATRIC DISORDER THAT IS DETRIMENTAL TO SOCIETY AND CURRENTLY HAS NO EFFECTIVE TREATMENTS. THE DIFFICULTY IN TREATING DRUG ADDICTION IS COMPOUNDED BY THE HIGH COMORBIDITY WITH OTHER PSYCHIATRIC ILLNESSES, INCLUDING ANXIETY DISORDERS. HERE, WE DEMONSTRATE THAT G9A, AN EPIGENETIC REPRESSOR OF GENE EXPRESSION, ACTING IN THE NUCLEUS ACCUMBENS, A BRAIN REWARD REGION, IS CAPABLE OF INCREASING BOTH ADDICTION- AND ANXIETY-LIKE BEHAVIORS IN RATS. THESE FINDINGS ARE INTRIGUING BECAUSE REPEATED COCAINE EXPOSURE DECREASES G9A IN THIS REGION AND THEREBY ENHANCES EXPRESSION OF CERTAIN ADDICTION-PROMOTING GENES. HOWEVER, OUR RESULTS SUGGEST THAT COUNTERING THIS COCAINE-INDUCED DECREASE IN G9A ACTIVITY ACTUALLY EXACERBATES ADDICTION AND SENSITIVITY TO RELAPSE UNDER STRESSFUL SITUATIONS. 2018 15 5804 33 STRAIN AND SEX DEPENDENT EFFECTS OF ISOLATION HOUSING RELATIVE TO ENVIRONMENTAL ENRICHMENT ON OPERANT SENSATION SEEKING IN MICE. SENSATION SEEKING IS A MULTIDIMENSIONAL PHENOTYPE THAT PREDICTS THE DEVELOPMENT OF DRUG ADDICTION IN HUMANS AND ADDICTION-LIKE DRUG SEEKING IN RODENTS. SEVERAL LINES OF EVIDENCE SUGGEST THAT CHRONIC STRESS INCREASES SENSATION SEEKING AND ADDICTION-LIKE DRUG SEEKING THROUGH COMMON GENETIC MECHANISMS. DISCOVERY AND CHARACTERIZATION OF THESE MECHANISMS WOULD REVEAL HOW CHRONIC STRESS INTERACTS WITH THE GENOME TO INFLUENCE SENSATION SEEKING AND HOW DRUGS OF ABUSE HIJACK THESE FUNDAMENTAL REWARD MECHANISMS TO DRIVE ADDICTION. TO THIS END, WE TESTED THE HYPOTHESIS THAT CHRONIC ISOLATION HOUSING STRESS (RELATIVE TO ENVIRONMENTAL ENRICHMENT) INFLUENCES OPERANT SENSATION SEEKING AS A FUNCTION OF STRAIN, SEX, OR THEIR INTERACTION. TO DETERMINE IF THE BXD RECOMBINANT INBRED PANEL COULD BE USED TO IDENTIFY GENETIC AND EPIGENETIC MECHANISMS UNDERLYING ANY IDENTIFIED GENE-BY-ENVIRONMENT INTERACTIONS, WE USED MICE FROM THE TWO BXD FOUNDER STRAINS. FOLLOWING 10 WEEKS OF DIFFERENTIAL HOUSING, WE ASSESSED OPERANT SENSATION SEEKING USING SEVERAL REINFORCEMENT SCHEDULES. THE PRIMARY FINDING FROM THIS STUDY WAS THAT DBA/2J BUT NOT C57BL/6J MICE WERE SIGNIFICANTLY VULNERABLE TO AN ISOLATION-INDUCED INCREASE (RELATIVE TO ENVIRONMENTAL ENRICHMENT) IN SENSATION SEEKING DURING EXTINCTION WHEN THE SENSORY REWARD WAS NO LONGER AVAILABLE; THIS EFFECT WAS SIGNIFICANTLY MORE ROBUST IN FEMALES. THESE DATA REVEAL A PREVIOUSLY UNKNOWN ISOLATION-INDUCED EFFECT ON EXTINCTION OF OPERANT SENSATION SEEKING THAT IS SEX-DEPENDENT, ADDICTION-RELEVANT, AND THAT CAN BE DISSECTED USING THE BXD RECOMBINANT INBRED PANEL. 2021 16 4945 27 PATERNAL PRECONCEPTION EVERY-OTHER-DAY ETHANOL DRINKING ALTERS BEHAVIOR AND ETHANOL CONSUMPTION IN OFFSPRING. ALCOHOL USE DISORDER IS A DEVASTATING DISEASE WITH A COMPLEX ETIOLOGY. RECENT PRECLINICAL STUDIES HAVE REVEALED THAT PATERNAL PRECONCEPTION CHRONIC INTERMITTENT ETHANOL (ETOH) EXPOSURE VIA VAPORIZED ETOH ALTERED DRINKING BEHAVIORS AND SENSITIVITY TO ETOH SELECTIVELY IN MALE OFFSPRING. IN THE CURRENT STUDY, WE USED A VOLUNTARY ORAL ROUTE OF PATERNAL PRECONCEPTION ETOH EXPOSURE, I.E., INTERMITTENT EVERY-OTHER-DAY TWO-BOTTLE CHOICE DRINKING, AND TESTED OFFSPRING FOR BEHAVIORAL ALTERATIONS. FIFTEEN ETOH DRINKING SIRES AND 10 CONTROL SIRES WERE MATED TO ETOH NAIVE FEMALES TO PRODUCE ETOH-SIRED AND CONTROL-SIRED OFFSPRING. THESE OFFSPRING WERE TESTED USING THE ELEVATED PLUS MAZE, OPEN FIELD, DRINKING IN THE DARK, AND UNLIMITED ACCESS TWO-BOTTLE CHOICE ASSAYS. WE FOUND THAT PATERNAL PRECONCEPTION EVERY-OTHER-DAY TWO-BOTTLE CHOICE DRINKING RESULTED IN REDUCED ETOH CONSUMPTION SELECTIVELY IN MALE OFFSPRING IN THE DRINKING IN THE DARK ASSAY COMPARED TO CONTROL-SIRED OFFSPRING. NO DIFFERENCES WERE DETECTED IN EITHER SEX IN THE UNLIMITED ACCESS TWO-BOTTLE CHOICE AND ELEVATED PLUS MAZE ASSAYS. OPEN FIELD ANALYSIS REVEALED COMPLEX CHANGES IN BASAL BEHAVIOR AND ETOH-INDUCED BEHAVIORS THAT WERE SEX SPECIFIC. WE CONCLUDED THAT PATERNAL PRECONCEPTION VOLUNTARY ETOH CONSUMPTION HAS PERSISTENT EFFECTS THAT IMPACT THE NEXT GENERATION. THIS STUDY ADDS TO A GROWING APPRECIATION THAT ONE'S BEHAVIORAL RESPONSE TO ETOH AND ETOH DRINKING BEHAVIOR ARE IMPACTED BY ETOH EXPOSURE OF THE PRIOR GENERATION. 2019 17 1086 35 COCAINE ADMINISTRATION AND ITS WITHDRAWAL ENHANCE THE EXPRESSION OF GENES ENCODING HISTONE-MODIFYING ENZYMES AND HISTONE ACETYLATION IN THE RAT PREFRONTAL CORTEX. CHRONIC EXPOSURE TO COCAINE, CRAVING, AND RELAPSE ARE ATTRIBUTED TO LONG-LASTING CHANGES IN GENE EXPRESSION ARISING THROUGH EPIGENETIC AND TRANSCRIPTIONAL MECHANISMS. ALTHOUGH SEVERAL BRAIN REGIONS ARE INVOLVED IN THESE PROCESSES, THE PREFRONTAL CORTEX SEEMS TO PLAY A CRUCIAL ROLE NOT ONLY IN MOTIVATION AND DECISION-MAKING BUT ALSO IN EXTINCTION AND SEEKING BEHAVIOR. IN THIS STUDY, WE APPLIED COCAINE SELF-ADMINISTRATION AND EXTINCTION TRAINING PROCEDURES IN RATS WITH A YOKED TRIAD TO DETERMINE DIFFERENTIALLY EXPRESSED GENES IN PREFRONTAL CORTEX. MICROARRAY ANALYSIS SHOWED SIGNIFICANT UPREGULATION OF SEVERAL GENES ENCODING HISTONE MODIFICATION ENZYMES DURING EARLY EXTINCTION TRAINING. SUBSEQUENT REAL-TIME PCR TESTING OF THESE GENES FOLLOWING COCAINE SELF-ADMINISTRATION OR EARLY (THIRD DAY) AND LATE (TENTH DAY) EXTINCTION REVEALED ELEVATED LEVELS OF THEIR TRANSCRIPTS. INTERESTINGLY, WE FOUND THE ENRICHMENT OF BRD1 MESSENGER RNA IN RATS SELF-ADMINISTERING COCAINE THAT LASTED UNTIL EXTINCTION TRAINING DURING COCAINE WITHDRAWAL WITH CONCOMITANT INCREASED ACETYLATION OF H3K9 AND H4K8. HOWEVER, DESPITE ELEVATED LEVELS OF METHYL- AND DEMETHYLTRANSFERASE-ENCODED TRANSCRIPTS, NO CHANGES IN GLOBAL DI- AND TRI-METHYLATION OF HISTONE H3 AT LYSINE 4, 9, 27, AND 79 WERE OBSERVED. SURPRISINGLY, AT THE END OF EXTINCTION TRAINING (10 DAYS OF COCAINE WITHDRAWAL), MOST OF THE ANALYZED GENES IN THE RATS ACTIVELY AND PASSIVELY ADMINISTERING COCAINE RETURNED TO THE CONTROL LEVEL. TOGETHER, THE ALTERATIONS IDENTIFIED IN THE RAT PREFRONTAL CORTEX MAY SUGGEST ENHANCED CHROMATIN REMODELING AND TRANSCRIPTIONAL ACTIVITY INDUCED BY EARLY COCAINE ABSTINENCE; HOWEVER, TO KNOW WHETHER THEY ARE BENEFICIAL OR NOT FOR THE EXTINCTION OF DRUG-SEEKING BEHAVIOR, FURTHER IN VIVO EVALUATION IS REQUIRED. 2017 18 4931 36 PATERNAL ALCOHOL EXPOSURE REDUCES ALCOHOL DRINKING AND INCREASES BEHAVIORAL SENSITIVITY TO ALCOHOL SELECTIVELY IN MALE OFFSPRING. ALCOHOL USE DISORDER (AUD) IS HERITABLE, BUT THE GENETIC BASIS FOR THIS DISEASE REMAINS POORLY UNDERSTOOD. ALTHOUGH NUMEROUS GENE VARIANTS HAVE BEEN ASSOCIATED WITH AUD, THESE VARIANTS ACCOUNT FOR ONLY A SMALL FRACTION OF THE TOTAL RISK. THE IDEA OF INHERITANCE OF ACQUIRED CHARACTERISTICS, I.E. "EPIGENETIC INHERITANCE," IS RE-EMERGING AS A PROVEN ADJUNCT TO TRADITIONAL MODES OF GENETIC INHERITANCE. WE HYPOTHESIZED THAT ALCOHOL DRINKING AND NEUROBIOLOGICAL SENSITIVITY TO ALCOHOL ARE INFLUENCED BY ANCESTRAL ALCOHOL EXPOSURE. TO TEST THIS HYPOTHESIS, WE EXPOSED MALE MICE TO CHRONIC VAPOR ETHANOL OR CONTROL CONDITIONS, MATED THEM TO ETHANOL-NAIVE FEMALES, AND TESTED ADULT OFFSPRING FOR ETHANOL DRINKING, ETHANOL-INDUCED BEHAVIORS, GENE EXPRESSION, AND DNA METHYLATION. WE FOUND THAT ETHANOL-SIRED MALE OFFSPRING HAD REDUCED ETHANOL PREFERENCE AND CONSUMPTION, ENHANCED SENSITIVITY TO THE ANXIOLYTIC AND MOTOR-ENHANCING EFFECTS OF ETHANOL, AND INCREASED BDNF EXPRESSION IN THE VENTRAL TEGMENTAL AREA (VTA) COMPARED TO CONTROL-SIRED MALE OFFSPRING. THERE WERE NO DIFFERENCES AMONG ETHANOL- AND CONTROL-SIRED FEMALE OFFSPRING ON THESE ASSAYS. ETHANOL EXPOSURE ALSO DECREASED DNA METHYLATION AT THE BDNFAEPROMOTER OF SIRE'S GERM CELLS AND HYPOMETHYLATION WAS MAINTAINED IN THE VTA OF BOTH MALE AND FEMALE ETHANOL-SIRED OFFSPRING. OUR FINDINGS SHOW THAT PATERNAL ALCOHOL EXPOSURE IS A PREVIOUSLY UNRECOGNIZED REGULATOR OF ALCOHOL DRINKING AND BEHAVIORAL SENSITIVITY TO ALCOHOL IN MALE, BUT NOT FEMALE, OFFSPRING. PATERNAL ALCOHOL EXPOSURE ALSO INDUCES EPIGENETIC ALTERATIONS (DNA HYPOMETHYLATION) AND GENE EXPRESSION CHANGES THAT PERSIST IN THE VTA OF OFFSPRING. THESE RESULTS PROVIDE NEW INSIGHT INTO THE INHERITANCE AND DEVELOPMENT OF ALCOHOL DRINKING BEHAVIORS. 2014 19 6639 37 UNRAVELING THE EPIGENOMIC AND TRANSCRIPTOMIC INTERPLAY DURING ALCOHOL-INDUCED ANXIOLYSIS. POSITIVE EFFECTS OF ALCOHOL DRINKING SUCH AS ANXIOLYSIS AND EUPHORIA APPEAR TO BE A CRUCIAL FACTOR IN THE INITIATION AND MAINTENANCE OF ALCOHOL USE DISORDER (AUD). HOWEVER, THE MECHANISMS THAT LEAD FROM CHROMATIN REORGANIZATION TO TRANSCRIPTOMIC CHANGES AFTER ACUTE ETHANOL EXPOSURE REMAIN UNKNOWN. HERE, WE USED ASSAY FOR TRANSPOSASE-ACCESSIBLE CHROMATIN FOLLOWED BY HIGH THROUGHPUT SEQUENCING (ATAC-SEQ) AND RNA-SEQ TO INVESTIGATE EPIGENOMIC AND TRANSCRIPTOMIC CHANGES THAT UNDERLIE ANXIOLYTIC EFFECTS OF ACUTE ETHANOL USING AN ANIMAL MODEL. ANALYSIS OF ATAC-SEQ DATA REVEALED AN OVERALL OPEN OR PERMISSIVE CHROMATIN STATE THAT WAS ASSOCIATED WITH TRANSCRIPTOMIC CHANGES IN THE AMYGDALA AFTER ACUTE ETHANOL EXPOSURE. WE IDENTIFIED A CANDIDATE GENE, HIF3A (HYPOXIA-INDUCIBLE FACTOR 3, ALPHA SUBUNIT), THAT HAD 'OPEN' CHROMATIN REGIONS (ATAC-SEQ PEAKS), ASSOCIATED WITH SIGNIFICANTLY INCREASED ACTIVE EPIGENETIC HISTONE ACETYLATION MARKS AND DECREASED DNA METHYLATION AT THESE REGIONS. THE MRNA LEVELS OF HIF3A WERE INCREASED BY ACUTE ETHANOL EXPOSURE, BUT DECREASED IN THE AMYGDALA DURING WITHDRAWAL AFTER CHRONIC ETHANOL EXPOSURE. KNOCKDOWN OF HIF3A EXPRESSION IN THE CENTRAL NUCLEUS OF AMYGDALA ATTENUATED ACUTE ETHANOL-INDUCED INCREASES IN HIF3A MRNA LEVELS AND BLOCKED ANXIOLYSIS IN RATS. THESE DATA INDICATE THAT CHROMATIN ACCESSIBILITY AND TRANSCRIPTOMIC SIGNATURES IN THE AMYGDALA AFTER ACUTE ETHANOL EXPOSURE UNDERLIE ANXIOLYSIS AND POSSIBLY PRIME THE CHROMATIN FOR THE DEVELOPMENT OF AUD. 2022 20 1116 27 COMPARATIVE ANALYSIS OF NEUROECTODERMAL DIFFERENTIATION CAPACITY OF HUMAN BONE MARROW STROMAL CELLS USING VARIOUS CONVERSION PROTOCOLS. HUMAN ADULT BONE MARROW-DERIVED MESODERMAL STROMAL CELLS (HMSCS) ARE ABLE TO DIFFERENTIATE INTO MULTIPLE MESODERMAL TISSUES, INCLUDING BONE AND CARTILAGE. THERE IS EVIDENCE THAT THESE CELLS ARE ABLE TO BREAK GERM LAYER COMMITMENT AND DIFFERENTIATE INTO CELLS EXPRESSING NEUROECTODERMAL PROPERTIES. THERE IS STILL DEBATE ABOUT WHETHER THIS RESULTS FROM CELL FUSION, ABERRANT MARKER GENE EXPRESSION OR REAL NEUROECTODERMAL DIFFERENTIATION. HERE WE EXTEND OUR WORK ON NEUROECTODERMAL CONVERSION OF ADULT HMSCS IN VITRO BY EVALUATING VARIOUS EPIGENETIC CONVERSION PROTOCOLS USING QUANTITATIVE RT-PCR AND IMMUNOCYTOCHEMISTRY. UNDIFFERENTIATED HMSCS EXPRESSED HIGH LEVELS OF FIBRONECTIN AS WELL AS SEVERAL NEUROECTODERMAL GENES COMMONLY USED TO CHARACTERIZE NEURAL CELL TYPES, SUCH AS NESTIN, BETA-TUBULIN III, AND GFAP, SUGGESTING THAT HMSCS RETAIN THE ABILITY TO DIFFERENTIATE INTO NEUROECTODERMAL CELL TYPES. PROTOCOLS USING A DIRECT DIFFERENTIATION OF HMSCS INTO A NEURAL PHENOTYPE FAILED TO INDUCE SIGNIFICANT CHANGES IN MORPHOLOGY AND/OR EXPRESSION OF MARKERS OF EARLY AND MATURE GLIAL/NEURONAL CELLS TYPES. IN CONTRAST, A MULTISTEP PROTOCOL WITH CONVERSION OF HMSCS INTO A NEURAL STEM CELL-LIKE POPULATION AND SUBSEQUENT TERMINAL DIFFERENTIATION IN MATURE GLIA AND NEURONS GENERATED RELEVANT MORPHOLOGICAL CHANGES AS WELL AS SIGNIFICANT INCREASE OF EXPRESSION LEVELS OF MARKER GENES FOR EARLY AND LATE NEURAL CELL TYPES, SUCH AS NESTIN, NEUROGENIN2, MBP, AND MAP2AB, ACCOMPANIED BY A LOSS OF THEIR MESENCHYMAL PROPERTIES. OUR DATA PROVIDE AN IMPETUS FOR DIFFERENTIATING HMSCS IN VITRO INTO MATURE NEUROECTODERMAL CELLS. NEUROECTODERMALLY CONVERTED HMSCS MAY THEREFORE ULTIMATELY HELP IN TREATING ACUTE AND CHRONIC NEURODEGENERATIVE DISEASES. ANALYSIS OF MARKER GENE EXPRESSION FOR CHARACTERIZATION OF NEURAL CELLS DERIVED FROM MSCS HAS TO TAKE INTO ACCOUNT THAT SEVERAL EARLY AND LATE NEUROECTODERMAL GENES ARE ALREADY EXPRESSED IN UNDIFFERENTIATED MSCS. 2006