Seite - 85 - in Advances in Neuroimmunology

BrainSci. 2016,6, 25 Subsequently,all ratsweresinglyhousedfor thedurationofexperimentalprocedures.Methodsused in this studywere approvedby InstitutionalAnimalCare andUseCommittee (IACUC, protocol number: 14-125)at theUniversityofNorthCarolina (ChapelHill,NC,USA). 2.2. LiquidDiet forControls and forChronicEthanolExposure In the initial experiment, rats that receivedanacuterestraint stresswereonchowdietbefore the stresschallenge. Forotherexperiments,anutritionally-completeandcalorically-balancedliquiddiet wasusedfor therats that receivedacontinuous7%(w/v) ethanoldiet followedby24hofwithdrawal (WCE)—anapproachpreviouslyutilizedbyour laboratory (e.g., [1,36,37]; seeFigure1) toadminister dailyethanoldosesof9–13g/kg.The liquidcontroldiet (CD)wascaloricallybalancedtotheWCE dietbyadjustmentsof theamountofdextrose. Ratswere fedeithercontroldietor theethanoldiet withamodifiedpair-feedingstrategy[1]. Figure1. Schematicsdepictingexperimentalprotocols.A: represents theacute restraint stress time course,whileB: representsthestressandwithdrawalfromchronicalcoholandC: representstheCRF1R antagonist study. 2.3. RestraintStress inControls andafterWithdrawal fromChronicEthanolExposure InitialeffortsdeterminedthetimecourseofstresseffectsonbraincytokinemRNAs. Stressconsisted of60minof restraint inplasticdecapicones. Theseratsweresacrificed2,4, 8, 24,or48hfollowingthe stress (seeschematic inFigure1A). 85