Effects Of Regulating Btn1a1 Expression On Lipid Synthesis In Mouse Spermatogoniaand Testicular Tissue

Lipid is an important component of sperm,and the dynamic stability of lipid provides basic conditions for spermatogenesis and maturation.Lipid synthesis is an important part of maintaining lipid homeostasis,which is closely related to testicular maturation,spermatogenesis,sperm motility and frost resistance.Butyrophilin subfamily 1 member a1(BTN1A1)is a member of the immunoglobulin(Ig)superfamily,and studies of lactation in cattle and mice have found that regulating the expression of this gene affected lipid droplet size and content,as well as intracellular lipid composition and content.However,the role of this gene in other mammalian tissues and cells has not been reported,especially in male germ cells and tissues,the expression and function are unknown.Therefore,this study first cloned the coding region of the mouse Btn1a1 gene and analyzed its expression in mouse tissues and spermatogonia.The effects of overexpression of Btn1a1 on lipid synthesis in mouse spermatogonia(GC-1)using lentiviral infection and inhibiting Btn1a1 expression in mouse testis on testicular morphology and lipid synthesis by RNA interference technology were investigated The results of the study will enrich our understanding of the Btn1a1 gene in lipid synthesis in the male reproductive system and provide a basis for further understanding of its relationship with semen freezing resistance.1.Mouse Btn1a1 gene coding region cloning and tissue expression profile analysis.We used PCR to amplify the coding region of Btn1a1 from mouse testicular tissue c DNA and sequenced it.Sequence analysis revealed that the coding region of mouse Btn1a1 gene was 1575 bp.The q RT-PCR results showed that Btn1a1 was expressed in the tissues and GC-1 cells of the examined mouse.The expression trend of Btn1a1 in testis and epididymis increased and then decreased with the growth and development of mouse,and remained high after sexual maturity;the expression of Btn1a1 in testis tissue was higher than that in epididymi tissues at the same developmental stage.2.Effect of modulating Btn1a1 expression on lipid synthesis in mouse spermatogonia and testicular tissue.We used lentiviral infection and si RNA injection to regulate the expression of Btn1a1 in GC-1 cells and testicular tissue,and respectively,to investigate its role on lipid synthesis.The results showed that overexpression of Btn1a1 increased lipid droplets,the content of partially saturated fatty acids and n-3 and n-6 unsaturated fatty acids in GC-1 cells,interference with Btn1a1 expression decreased testicular lipid content.The q RT-PCR results showed that compared with the control group,the Btn1a1 overexpression group significantly increased the expression of Fasn,Plin2,Scd1 and fatty acid de novo synthesis related genes,Acaca,Srebp-1 and Pparγ were significantly increased(P<0.05).After interfering with testicular Btn1a1 gene expression,compared with the control group,the experimental group significantly decreased the expression of lipid synthesis genes,Fasn and Pparγ(P<0.05),and extremely significantly decreased the expression of Plin2(P<0.01).The above results indicate that Btn1a1 expressed in all major tissues of mouse,and its expression in mouse testis and epididymis was maintained at high levels after sexual maturation.Btn1a1 may regulate the process of lipid synthesis by regulating the expression of genes related to lipid synthesis,thereby maintaining spermatogonial and testicular lipid homeostasis.