The aims of this study were to establish whether the sex-determining region Y gene and its mRNA transcript are present in the Y sperm and X sperm of bulls and, if present, determine their cellular localization. Semen was collected from three bulls and sorted by flow cytometry into X- and Y-chromosome populations. Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine Sry mRNA expression in X sperm and Y sperm. The presence and localization of Sry DNA and RNA were investigated by fluorescence in situ hybridization (FISH). RT-PCR detected a single Sry transcript of 142 bp in Y sperm but not in X sperm. In Y sperm, the FISH-positive rates for Sry DNA and Sry RNA did not differ significantly from the re-analyzed Y sperm purity. In further experiments, there were no significant differences between the FISH-positive rate for Sry RNA and the re-analyzed Y sperm purity for X-sorted, Y-sorted, or unsorted sperm. In conclusion, FISH analysis revealed that Sry transcripts are present at the edges of the sperm heads of Y sperm but are absent from X sperm.
CapraE.TurriF.LazzariB.CremonesiP.GliozziT.M.FojadelliI.StellaA. & PizziF. (2017) Small RNA sequencing of cryopreserved semen from single bull revealed altered miRNAs and piRNAs expression between high- and low-motile sperm populations. BMC Genomics1814.
DewingP.ChiangC.W.SinchakK.SimH.FernagutP.O.KellyS.ChesseletM.F.MicevychP.E.AlbrechtK.H.HarleyV.R. & VilainE. (2006) Direct regulation of adult brain function by the male-specific factor SRY. Curr. Biol.16415-420.
HanleyN.A.HaganD.M.Clement-JonesM.BallS.G.StrachanT.Salas-CortesL.McElreaveyK.LindsayS.RobsonS.BullenP.OstrerH. & WilsonD.I. (2000) SRY, SOX9, and DAX1 expression patterns during human sex determination and gonadal development. Mech. Dev.91403-407.
LambardS.Galeraud-DenisI.MartinG.LevyR.ChocatA. & CarreauS. (2004) Analysis and significance of mRNA in human ejaculated sperm from normozoospermic donors: relationship to sperm motility and capacitation. Mol. Hum. Reprod.10535-541.
UenoA.TakedaM.HirasawaK.ItagakiS. & DoiK. (1997) Detection of viral RNA by electron microscopic in situ hybridization (ISH-EM) in the germinal epithelium of mice infected with encephalomyocarditis (EMC) virus. Exp. Anim.4679-81.
WilhelmD.MartinsonF.BradfordS.WilsonM.J.CombesA.N.BeverdamA.BowlesJ.MizusakiH. & KoopmanP. (2005) Sertoli cell differentiation is induced both cell-autonomously and through prostaglandin signaling during mammalian sex determination. Dev. Biol.287111-124.