Identification,Tissue Distribution Of Vulpes Beta-Defensins And Their Expression In Reproductive Tissue Of Asthenozoospermia Foxes

Beta-defensin is an important cationic antimicrobial peptide molecule in mammals and plays an important role in innate immunity and acquired immunity.In recent years,β-defensins have been found to be relatively concentrated in the male reproductive system of mammals,and play an important role in regulating the functional activity of sperm and participating in the effective reproduction of animals.At present,studies on mammalian β-defensins are more common in animals such as humans,mice and macaques,and studies on vulpes beta-defensins(vBDs)have not been reported.In this study,the blue fox and red fox vBD1,vBD108 and vBD122 genes were obtained by PCR and the distribution of 10 vBDs was identified.qPCR was used to study the expression levels of blue fox vBD1,vBD2,vBD3,vBD108 and vBD122 genes and the expression levels of vBD1,vBD 108 and vBD 122 were compared with those of red fox.qPCR and immunohistochemical techniques were used to study the gene expression level and tissue localization of vBD108 and vBD122 in the reproductive tissues of non-obstructive asthenozoospermia(NOA)blue fox.The results are as follows:(1)The full CDS coding region genes of vBD1,vBD108 and vBD122 were successfully obtained.The full length of the CDS coding region was 207 bp,216 bp and 195 bp,encoding 68,71 and 64 amino acids,respectively.The signal peptide lengths were 23,19 and 22 respectively.The amino acids have the closest homology to the canine β-defensin gene;the blue foxes vBD1,vBD108 and vBD122 have 1,3 and 2 amino acid site mutations,respectively,and the red foxes vBD1,vBD108 and vBD122 have 2 amino acid sequences,respectively.,2 and 1 amino acid site mutations.(2)The 10 vBDs genes were irregularly distributed in the tissues of blue foxes and red foxes.vBD3,vBD108 and vBD 122 were mainly expressed in testis and epididymis tissues,and vBD1 and vBD2 were mainly expressed in liver and kidney tissues.In blue foxes’ tissues,the expression of vBD1 in kidney and liver was significantly higher than that in vBD2(P<0.05).The expression of vBD 122 in testis was significantly higher than that in vBD3 and vBD108.The expression of vBD108 in epididymis was significantly higher than that in vBD3.And vBD122(P<0.05);and the expression of vBD1 in the kidney and liver tissues of blue foxes,and the expression of vBD108 and vBD122 in testis and paraffin tissues were higher than those in red foxes(P<0.01).(3)The expression of vBD108 mRNA in the reproductive tissues of NOA blue foxes was:testis>epididymal head>epididymis>epididymal tail(P<0.05),and the expression level of vBD108 mRNA in NOA blue foxes was significantly lower than that in the health group.The difference in the structure of the epididymis was the largest;The expression of vBD122 mRNA in the reproductive tissues of NOA blue foxes was:testis>epididymal head>epididymis>epididymal tail(P<0.05),and the expression of vBD122 in the reproductive tract of NOA blue foxes was significantly lower than health group,which has the largest difference in testicular tissue;vBD108 protein was mainly distributed in testicular interstitial cells,spermatogenic cells,supporting cells and epithelial pseudostratified epithelial columnar ciliated cells.The positive signal strength was:epididymis Body>testis>epididymis head>epididymal tail,vBD108 protein positive signal of NOA foxes is weak;vBD122 protein is mainly distributed in testicular spermatogenic cells,supporting cells,and epididymal pseudostratified epithelial columnar ciliated cells.There is basically no signal in the cytoplasmic cells,and the positive signal strength is:testis>epididymal head>epididymis>epididymal tail,weak positive signal in the reproductive tissue of NOA foxes.