ART3调控小鼠精子发生的作用与机制研究

Study on the Effects and Mechanism of ART3 in Regulating Spermatogenesis in Mice

为探究ADP核糖基转移酶3(ADP-ribosyl transferase 3,ART3)在精子发生及变形中的作用及机制,本研究随机选取21只6~8周龄健康ICR雄鼠分为对照组、ART3抑制剂3-甲氧基苯甲酰胺(3-methoxybenzamide,3-MBA)处理组,每组各3个重复。处理组睾丸注射3-MBA后12 h、24 h、48 h、5 d、7 d、14 d采集睾丸和附睾组织。跟踪分析其中的精子发生过程发现,抑制ART3导致睾丸萎缩、附睾尾精子密度降低、畸形率升高。组织学观察发现,曲细精管生精细胞逐渐减少,且排列松散,部分生精细胞脱落至管腔;附睾头和附睾尾出现非精子细胞成分,经顶体蛋白免疫荧光鉴定非精子细胞成分为圆形精细胞。进一步免疫荧光检测发现,抑制剂处理降低了精细胞和支持细胞间的细胞粘附蛋白Nectin-3水平,曲细精管出现生精细胞凋亡现象,AKT信号通路相关蛋白水平下降。据此推测,ART3通过调控精细胞与支持细胞间连接,影响AKT通路的抗凋亡作用,促进精细胞变形。

The study aimed to investigate the role and mechanism of ADP-ribosyl transferase 3(ART3)in spermatogenesis and deformation.In this study,21 healthy ICR male mices at 6-8 weeks were randomly selected and divided into control group,ART3 inhibitor 3-MBA treatment groups,with 3 replicates in each group.Testicular and epididymis tissues were collected for 12 h,24 h,48 h,5 d,7 d,and 14 d after testicular injection of 3-MBA in the treatment groups.Analysis of spermatogenesis in them showed that inhibition of ART3 resulted in testicular atrophy,decreased sperm density in the cauda epididymidis,and increased sperm deformity rate.Histological observation showed that the spermatogenic cells in the seminiferous tubules were gradually reduced and loosely arranged,and some of them were detached to the lumen;non-sperms components appeared in the caput and cauda epididymidis.The components were identified as round spermatids by immunofluorescence of acrosomal proteins.Further immunofluorescence showed that the inhibitor treatment reduced the level of Nectin-3,a cell adhesion protein between spermatids and sertoli cells,led to the apoptosis of spermatids and the expression of AKT signaling pathway-related proteins decreased.It is hypothesised that ART3 promotes round spermatids deformation by modulating spermatids-sertoli cells junctions and affecting the anti-apoptotic effect of AKT pathway.