不同日龄犊牛睾丸中细胞周期调控基因CyclinB1和p34^(cdc2)表达的差异

Expressions of Cell Cycle Regulation Genes CyclinB1 and p34^(cdc2) in the Testes of Calves Aged on Different Days

旨在探究不同日龄犊牛睾丸中细胞周期调控基因CyclinB1和p34cdc2的表达情况。本试验选取发育正常、体重相似的5~6d荷斯坦哺乳公犊18头,随机分为3组,每组6头,饲喂相同的基础日粮,分别在30(断奶)、60和90d时从各组分别随机抽取2头,采集其睾丸备用;通过qRT-PCR技术检测不同日龄犊牛睾丸中细胞周期调控基因CyclinB1和p34cdc2 mRNA表达规律,免疫组化技术对睾丸CyclinB1和p34cdc2进行定位分析。结果表明:90d时CyclinB1和p34cdc2 mRNA表达量显著高于30与60d(P<0.05),但30与60d两种基因mRNA表达量均差异不显著(P>0.05);p34cdc2蛋白在不同日龄犊牛表达差异显著(P<0.05);CyclinB1蛋白在60、90d的表达量显著高于30d(P<0.05),但60和90d差异不显著(P>0.05)。综上表明,CyclinB1和p34cdc2 mRNA和蛋白在睾丸的表达量随着犊牛日龄的增加而增加,且不同的日龄CyclinB1和p34cdc2 mRNA和蛋白表达量存在一定的差异。

This study was performed to investigate the expression of cell cycle regulation genes CyclinB1 and p34cdc2 in testis of calves at different ages.The eighteen 5-6days healthy Holstein calves(with similar weight)were randomly divided into 3groups and fed with the similar basal diet.The testes of 2calves from each group were collected at the time of 30,60 and 90drespectively.QRT-PCR method was used to detect the mRNA expression of the cell cycle regulatory genes CyclinB1 and p34cdc2 in testes of calves at different ages.The location of CyclinB1 and p34cdc2 in the testes was detected by immunohistochemical methods.The results showed that the highest expression of CyclinB1 and p34cdc2 mRNA and protein was obtained in the calves' testes of 90 d.The mRNA and protein expression levels of CyclinB1 and p34cdc2 increased with the increasing age.The mRNA expression level of the 2genes at the age of 90 d were significantlyhigher than that at 30 and 60d(P<0.05).However,no significant difference was observed between 30 and 60d(P>0.05).The protein expression of p34cdc2 in the testes of calves at different ages was significant(P<0.05).Although the protein expression of CyclinB1 at 60and 90 d had no significant difference(P>0.05),they were significantly higher than those at 30 d.This study indicated that the expression of CyclinB1 and p34cdc2 mRNA and protein increased with the increasing age and CyclinB1 and p34cdc2 in the testes of calves at different ages was significant.