SLXL1, a Novel Acrosomal Protein, Interacts with DKKL1 and Is Involved in Fertilization in Mice 英文参考文献.docVIP

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SLXL1, a Novel Acrosomal Protein, Interacts with DKKL1 and Is Involved in Fertilization in Mice 英文参考文献.doc

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SLXL1, a Novel Acrosomal Protein, Interacts with DKKL1 and Is Involved in Fertilization in Mice 英文参考文献

SLXL1,aNovelAcrosomalProtein,InteractswithDKKL1 andIsInvolvedinFertilizationinMice Xin-jieZhuang1,2,Xiao-junHou2,Shang-YingLiao2,Xiu-XiaWang2,HowardJ.Cooke3,MingZhang1, ChunshengHan2* 1StateKeyLaboratoryforConservationandUtilizationofSubtropicalAgro-bioresources,AnimalReproductionInstitute,GuangxiUniversity,Nanning,Guangxi,People’s RepublicofChina,2StateKeyLaboratoryofReproductiveBiology,InstituteofZoology,ChineseAcademyofSciences,Beijing,People’sRepublicofChina,3Instituteof GeneticandMolecularMedicineMRCHumanGeneticsUnit,WesternGeneralHospital,Edinburgh,Scotland,UnitedKingdom Abstract Background:Spermatogenesisisacomplexcellulardevelopmentalprocesswhichinvolvesdiversefamiliesofgenes.TheXlr (X-linked,lymphocyteregulated)familyincludesmultiplemembers,onlyafewofwhichhavereportedfunctionsinmeiosis, post-meioticmaturation,andfertilizationofgermcells.Slx-like1(Slxl1)isamemberoftheXlrfamily,whoseexpressionand functioninspermatogenesisneedtobeelucidated. Methodology/PrincipalFindings:ThemRNAandproteinexpressionandlocalizationofSlxl1wereinvestigatedbyRT-PCR, Westernblottingandimmunohistochemistryindifferenttissuesandatdifferentstagesofspermatogenesis.Theinteracting partner of SLXL1 was examined by co-immunoprecipitation and co-localization. Assessment of the role of SLXL1 in capacitation,acrosomereaction,zonapellucidabinding/penetration,andfertilizationwascarriedoutinvitrousingblocking antisera.TheresultsshowedthatSlxl1mRNAandproteinwerespecificallyexpressedinthetestis.SLXL1wasexclusively located in the acrosome of post-meiotic germ cells and interacts with DKKL1 (Dickkopf-like1), which is an acrosome- associated protein and plays an important role in fertilization. The rates of zona pellucida binding/penetration and fertilizationweresignificantlyreducedbytheanti-SLXL1polyclonalantiserum. Conclusions/Significance: SLXL1isthefirstidentifiedmemberoftheXLRfamilythatisassociatedwithacrosomeandis involvedinzonapellucidbinding/penetrationandsubsequentfertilization.Theseresults,