目的:SPAG9作为MAPK家族中的成员,在精卵融合过程中发挥着重要的作用。本研究检测SPAG9在人精液精子中的表达情况。方法:取人不同组织(肌肉、肝、食管、肺、胃、。肾、前列腺、子宫、睾丸、附睾)标本及健康男性的精液精子,为了排除圆形细胞的污染,精液标本经非连续性Percoll梯度离心、分离纯化;采用逆转录.聚合酶链式反应(RT—PCR)和间接免疫荧光方法,从mRNA和蛋白水平检测SPAG9在精子中的表达。结果:RT—PCR结果显示,SPAG9mRNA不仅在各种不同的组织中有表达,而且在人精液精子中也有表达;间接免疫荧光结果显示,SPAG9蛋白主要定位于人精液精子头部的赤道板和精子尾部鞭毛上。结论:SPAG9在人精液精子中有表达,可能在精子获能和运动中有一定作用。
Objective: SPAG9, as a member of the MAPK family, plays an important role in sperm-egg fusion. This study aimed to detect the expression of SPAG9 in human ejaculated spermatozoa. Methods : Different human tissues ( as from the muscle, liver, esophagus, lung, stomach, kidney, prostate, uterus, testis and epididymis) and semen samples were obtained from healthy volunteers, and semen analyses were performed according to the WHO criteria. Human ejaculated spermatozoa were purified by discontinuous Percoil density gradient centrifugation to rule out the contamination of germ cells and leucocytes. RT-PCR and indirect immunofluorescence were used to detect the expression of SPAG9 in human spermatozoa. Results: RT-PCR showed that SPAG9 mRNA was expressed in different tissues and human ejaculated spermatozoa. Indirect immunofluorescence studies revealed the location of SPAG9 protein in the equatorial plate and flagella of human spermatozoa. Conclusion : SPAG9 is expressed in ejaculated spermatozoa and may play a role in sperm capacitation and motility.