Expression and localization of forkhead transcriptional factor 2 (Foxl2) in the gonads of protogynous wrasse, Halichoeres trimaculatus.docVIP

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Expression and localization of forkhead transcriptional factor 2 (Foxl2) in the gonads of protogynous wrasse, Halichoeres trimaculatus.doc

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Expression and localization of forkhead transcriptional factor 2 (Foxl2) in the gonads of protogynous wrasse, Halichoeres trimaculatus

Kobayashietal.BiologyofSexDifferences2010,1:3 /content/1/1/3 RESEARCH OpenAccess Expressionandlocalizationofforkhead transcriptionalfactor2(Foxl2)inthegonadsof protogynouswrasse,Halichoerestrimaculatus YasuhisaKobayashi1,2*,RyoHoriguchi1,2,RyoNozu1,MasaruNakamura1,2 Abstract Background:Three-spotwrasse,Halichoerestrimaculatus,isamarineprotogynoushermaphroditefish.Individuals matureeitherasinitialphase(IP)malesorfemales.AppropriatesocialcuesinducethesexchangefromIPfemale toterminalphase(TP)male.However,themolecularmechanismsbehindsuchasexchangeremainlargely unknown.Recently,theforkheadtranscriptionfactor2(Foxl2)wasidentifiedasanessentialregulatorofvertebrate ovariandevelopment/function/phenotype.Inspiredbythisinformation,wecharacterizedtheexpressionpatternsof Foxl2intheprotogynouswrasseassumingFoxl2asthefemale-specificmarkerinthisspecies. Methods:First,weclonedFoxl2cDNAfromovarybyreversetranscriptionpolymerasechainreaction(RT-PCR) followedbyrapidamplificationofcDNAends(RACE).Next,weanalysedexpressionpatternofFoxl2messenger RNA(mRNA)andproteiningonadsofdifferentsexualphasesbyrealtimequantitativePCRassayandflour fluorescenceimmunohistochemicalmethod,respectively.Additionally,westudiedthechangesinFoxl2expression patternduringaromataseinhibitor(AI)-inducedsexchange. Results:Theaminoacidsequence(306AA)ofwrasseFoxl2,especiallytheforkheaddomain,showshighidentity withthatofotherreportedteleostFoxl2s.Quiteunexpectedly,nosexualdimorphismwasobservablebetweenthe testesandovaryintheexpressionpatternofFoxl2.Infemalephasefish,signalsforFoxl2proteinweredetectable inthegranulosacells,butnotthethecacells.TranscriptlevelsofFoxl2inthetestesofIPandTPmaleswere identicaltothatintheovariesoffemalesand,further,Foxl2proteinwasfoundtobelocalizedintheinterstitial cellsincludingtubulesandLeydigcells.TreatmentwithAIinducedsexchangeinmalegonadsandan up-regulationwasseenintheexpressionofFoxl2inthesegonads. Conclusions:Unlikeinothervertebrates,includingteleosts,Foxl2mayhaveadifferentroleinthenatur