Action Mechanism of Inhibin α-Subunit on the Development of Sertoli Cells and First Wave of Spermatogenesis in Mice 英文参考文献.docVIP

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Action Mechanism of Inhibin α-Subunit on the Development of Sertoli Cells and First Wave of Spermatogenesis in Mice 英文参考文献.doc

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Action Mechanism of Inhibin α-Subunit on the Development of Sertoli Cells and First Wave of Spermatogenesis in Mice 英文参考文献

ActionMechanismofInhibina-Subunitonthe DevelopmentofSertoliCellsandFirstWaveof SpermatogenesisinMice KailaiCai1,GuohuaHua1,SibtainAhmad1,2,AaixinLiang1,LiHan1,CanjieWu1,FeifeiYang1 ,Liguo Yang1* 1KeyLaboratoryofAgriculturalAnimalGenetics,BreedingandReproduction,EducationMinistryofChina,HuazhongAgriculturalUniversity,Wuhan,People’sRepublicof China,2DepartmentofLivestockManagement,UniversityofAgriculture,Faisalabad,Pakistan Abstract InhibinisanimportantmarkerofSertolicell(SC)activityinanimalswithimpairedspermatogenesis.However,theprecise relationship between inhibin and SC activity is unknown. To investigate this relationship, we partially silenced both the transcriptionandtranslationofthegeneforthea-subunitofinhibin,Inha,usingrecombinantpshRNAvectorsdeveloped withRNAi-ReadypSIREN-RetroQ-ZsGreenVector(ClontechLaboratories,MountainView,Calif).WefoundthatInhasilencing suppresses the cell-cycle regulators Cyclin D1 and Cyclin E and up-regulates the cell-cycle inhibitor P21 (as detected by Westernblotanalysis),therebyincreasingthenumberofSCsintheG1phaseofthecellcycleanddecreasingtheamountin theS-phaseofthecellcycle(asdetectedbyflowcytometry).InhasilencingalsosuppressedPdgfa,Igf1,andKitlmRNAlevels and up-regulated Tgfbrs, Inhba, Inhbb, Cyp11a1, Dhh, and Tjp1 mRNA levels (as indicated by real-time polymerase chain reaction[PCR]analysis).ThesefindingsindicatethatInhahasthepotentialtoinfluencetheavailabilityoftheligandinhibin and its antagonist activin in the SC in an autocrine manner and inhibit the progression of SC from G1 to S. It may also participateinthedevelopmentoftheblood–testisbarrier,Leydigcells,andspermatogenesisthroughitseffectonDhh,Tjp1, Kitl,andPdgfa.Real-timePCRandWesternblotanalysesofInha,Inhba,andInhbbmRNAandInhalevelsovertimeshowthat Inhaplaysanimportantroleintheformationofroundspermatidduringthefirstwaveofspermatogenesisinmice. Citation:CaiK,HuaG,AhmadS,LiangA,HanL,etal.(2011)ActionMechanismofInhibina-SubunitontheDevelopmentofSertoliCellsandFirstWaveof S