Ces5a基因在大鼠睾丸中的表达

目的:分析羧酸酯酶5a (Ces5a)基因在大鼠睾丸发育过程中的表达情况。方法:选取3组不同窝别出生后2、4、6、8、10、12、14、15、16、20、25、29、30、31、35、40、45、50、55、60、65 d 21个时间点的大鼠,采用RT-PCR、Western印迹、免疫组化、HE染色分别检测大鼠睾丸组织中Ces5a基因在mRNA转录水平和蛋白表达量及定位。结果:①Ces5a mRNA在大鼠出生后的第2~65天均有表达,大鼠出生后2~12 d Ces5a mRNA的表达量较低;与2~12 d相比,14~16 d的表达量下降至最低水平(P<0.05);与14~16 d相比,20~35 d该表达量有显著升高(P<0.05);与20~35 d相比,40~65 d的表达量有显著升高,达到高峰(P<0.05)。②Ces5a蛋白在大鼠出生后第2~65天均有表达,在大鼠出生后2~12 d Ces5a蛋白的表达量较低;与2~12 d相比,14~16 d Ces5a蛋白的表达量无显著变化(P>0.05);与14~16 d相比,20~35 d的表达量有显著升高(P<0.05);与20~35 d相比,40~65 d的表达量无显著变化(P>0.05)。③Ces5a蛋白表达于精原细胞、精母细胞及圆形精子细胞。结论:①Ces5a基因可能参与了大鼠精原细胞增殖过程及减数分裂过程;②Ces5a基因可能在圆形精子发生和精子变形过程中发挥特殊的作用。

An epididymis-specific carboxyl esterase CES5A is required for sperm capacitation and male fertility in the rat

Despite the fact that the phenomenon of capacitation was discovered over half century ago and much progress has been made in identifying sperm events involved in capacitation, few specific molecules of epididymal origin have been identified as being directly involved in this process in vivo. Previously, our group cloned and characterized a carboxyl esterase gene CesSa in the rat epididymis. The CES5A protein is mainly expressed in the corpus and cauda epididymidis and secreted into the corresponding lumens. Here, we report the function of CESSA in sperm maturation. By local injection of Lentivirus-mediated siRNA in the CESSA.expressing region of the rat epididymis, CesSa-knockdown animal models were created. These animals exhibited an inhibited sperm capacitation and a reduction in male fertility. These results suggest that CESSA plays an important role in sperm maturation and male fertility.