The Characteristic Analysis Of CLDN3 Gene In Yak (Bos Grunniens) And The Effects Of FSH On CLDN3 CLDH11 Expression In Sertoli Cells

The CLDNs of sertoli cells(SCs)regulate the progress of opening or closing the microenviroment during the Germ Cell Migration Across Sertoli Cell Tight Junctions.In order to explore the biological effects of CLDNs on yak SCs,this study used reverse transcription polymerase chain reaction(RT-PCR)to clone CLDN3 cDNA of yak(Bos grunniens)and used bioinformatics method to analyze it’s biological characteristics.Then the effected of follicle-stimulating hormone(FSH)to yak SCs on CLDN3 mRNA,CLDN11 mRNA in vitro were evaluated by quantitative real time polymerase chain reaction(qRT-PCR).The effected of CLDN3 CLDN11 protein in yak with FSH were dected by Western bloting(WB).What’s more,the location of CLDN3 protein,CLDN11 protein on SCs were detected by immunofluorescence.The results showed that:(1)The yak SCs exhibited bipolar corpuscula in nucleus after Feulgen staining and had a positive reaction of Fasl by the immunocytochemical identification.The CLDN3 cDNA of yak was successfully cloned,and it contained ORF of 1074 bp length(GenBank No.YK411920)and 660 bp CDs,encoding 219 amino acids.The sequence alignment showed that the homology between yak and other species were relatively high(≥90%).(2)qRT-PCR detection showed that the CLDN3 mRNA,CLDN11 mRNA of 25 ng/mL group was the highest expression level(p<0.01).(3)The 25 ng/m L FSH group of CLDN3 protein was the highest of the groups(p<0.01),The expression of CLDN11 protein in 25 ng/m L FSH was higher than that in the control group(p>0.05).(4)The CLDN3 protein could be detected in nucleus,cytoplasm and plasma membrane of SCs from immunofluorescence analysis.And the CLDN11 protein could be detected in cytoplasm and plasma membrane of SCs from immunofluorescence analysis.In summary,the CLDN3 amino acids sequence of yak is conserved in the different genus.FSH may be as an important hormone during the formation of TJ in testis of yak.The expression of CLDN3 and CLDN11 rely on concentration,time of FSH.The research provided basis for revealing the biological effect of yak CLDN3 CLDN11 during the germ cell migration across sertoli cell tight junctions.