Proteomic Changes in Rat Spermatogenesis in Response to In Vivo Androgen Manipulation; Impact on Meiotic Cells 英文参考文献.docVIP

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Proteomic Changes in Rat Spermatogenesis in Response to In Vivo Androgen Manipulation; Impact on Meiotic Cells 英文参考文献.doc

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Proteomic Changes in Rat Spermatogenesis in Response to In Vivo Androgen Manipulation; Impact on Meiotic Cells 英文参考文献

ProteomicChangesinRatSpermatogenesisinResponse toInVivoAndrogenManipulation;ImpactonMeiotic Cells PeterG.Stanton1,2*.,PavelSluka1.¤,CarolineF.H.Foo1,AndrewN.Stephens1,A.IanSmith2, RobertI.McLachlan1,3,LizaO’Donnell1,4 1PrinceHenry’sInstitute,MonashMedicalCentre,Clayton,Victoria,Australia,2DepartmentofBiochemistryandMolecularBiology,MonashUniversity,Clayton,Victoria, Australia,3DepartmentofObstetricsandGynaecology,MonashUniversity,Clayton,Victoria,Australia,4DepartmentofAnatomyandDevelopmentalBiology,Monash University,Clayton,Victoria,Australia Abstract Theproductionofmaturespermisreliantonandrogenactionwithinthetestis,anditiswellestablishedthatandrogensact onreceptorswithinthesomaticSertolicellstostimulatemalegermcelldevelopment.MicelackingSertolicellandrogen receptors(AR)showlatemeioticgermcellarrest,suggestingSertolicellstransducetheandrogenicstimulusco-ordinating thisessentialstepinspermatogenesis.Thisstudyaimedtoidentifygermcellproteinsresponsivetochangesintesticular androgenlevelsandtherebyelucidatemechanismsbywhichandrogensregulatemeiosis.Testicularandrogenlevelswere suppressedfor9weeksusingtestosteroneandestradiol-filledsilasticimplants,followedbyashortperiodofeitherfurther androgen suppression (via an AR antagonist) or the restoration of intratesticular testosterone levels. Comparative proteomics were performed on protein extracts from enriched meiotic cell preparations from adult rats undergoing androgendeprivationandreplacementinvivo.Lossofandrogenicstimuluscausedchangesinproteinswithknownrolesin meiosis (including Nasp and Hsp70–2), apoptosis (including Diablo), cell signalling (including 14-3-3 isoforms), oxidative stress, DNA repair, and RNA processing. Immunostaining for oxidised DNA adducts confirmed spermatocytes undergo oxidative stress-induced DNA damage during androgen suppression. An increase in PCNA and an associated ubiquitin- conjugating enzyme (Ubc13) suggested a role for PCNA-mediated regulation of DNA repair pathways in spermatocytes. Cha