小鼠子宫颈部位输卵管蛋白表达的初步研究

用RT-PCR及免疫印迹法检测证实小鼠子宫颈粘膜上皮细胞具表达输卵管蛋白的能力,宫颈部所获得的输卵管蛋白转录产物, 310到 714的405bp的cDNA片段经序列测定及blast比较表明与输卵管部位表达的输卵管蛋白完全一致(100%)。小鼠子宫颈输卵管蛋白在动情周期的表达波动情况与输卵管粘膜上皮的类似,其表达于小鼠动情期明显增加。Westernblot显示小鼠子宫颈提取物中存在两种不同分子量(60KD,30KD)的输卵管蛋白。原位杂交结果则提示子宫颈粘膜上皮细胞含丰富的输卵管蛋白mRNA信号。通过"原体中风"实验未发现输卵管蛋白在体外对精子活动有影响,其功能尚须进一步分析。

PRELIMINARY STUDY ON EXPRESSION OF OVIDUCTIN IN MOUSE CERVIX

In the present study,in order to investigate the pattern of the expression and location of oviductin using RT-PCR and Western blot,a 410bp cDNA fragment was amplified from mouse cervix tissue. Sequencing analysis and homology comparison with BLAST alignment showed that this fragment amplified from mouse cervix was homologous to the region from 310 to 714 of the mouse OGP gene in Genebank by the degree up to 100%. Western blot showed 60 KD and 30 KD components from mouse cervix same as those from mouse oviduct mucosa epithelium. Semi-quantitative RT-PCR analysis showed that expression of OGP was significantly higher at the estrous stage than that of non-estrous stage and almost no expression in the immature mouse. Those results suggest that the mRNA level of mouse cervix oviductin was related to estrous cycle,implied with estrogen-dependence. To further confirm the results,in situ hybridization histochemistry analysis showed that OGP mRNA was expressed at columnar epithelium of mouse cervix. To clarify the possible significance of oviductin expressed at the cervix,the function of oviductin on the mobility of human sperm was studied using "loss-of-function". The VAP?VCL?ALH?VSL showed no significant differences of the mobility of the human sperm in the conditioned medium blocked by the anti-CTP rOGP(rabbit Oviductin) antiserum compared with that in the conditioned medium of the normal rabbit serum. These results suggest that rOGP may not affect the mobility of human sperm in vitro.