TRPV6在蛋鸡不同生殖器官组织的表达

Expression of TRPV6 in Different Reproductive Organs in Laying Hens

【目的】研究瞬时性受体电位通道香草酸受体6(transient receptor potential vanilloid receptor6,TRPV6)在蛋鸡卵巢、输卵管的膨大部、峡部以及子宫部的表达分布。【方法】本试验采用RT-PCR和免疫组化技术研究TRPV6在蛋鸡不同生殖器官的定性表达,同时采用Real-time PCR和Western blotting方法观察其表达量的变化。【结果】在卵巢,TRPV6蛋白主要分布在初级卵泡和次级卵泡。在输卵管的膨大部、峡部以及子宫部,TRPV6主要分布在黏膜上皮的纤毛上皮细胞游离端(面向管腔),其中子宫部黏膜上皮的基底层也有少量分布。另外,与卵巢组织相比,膨大部和峡部TRPV6 mRNA表达量显著降低(P<0.05)。同时,膨大部TRPV6蛋白含量也显著降低(P<0.05),子宫部组织TRPV6 mRNA水平低于卵巢,蛋白水平则相反,但均无统计学差异(P>0.05)。【结论】TRPV6在卵巢、输卵管的膨大部、峡部以及子宫部均有表达,且卵巢和输卵管的子宫部表达量较高,提示TRPV6可能参与卵泡的发育成熟,对蛋鸡输卵管的子宫部Ca2+转运也具有重要意义。

[Objective] The aim of this study was to investigate the localization and expression of transient receptor potential vanilloid receptor 6 （TRPV6） in the ovary, magnum, isthmus and uterus parts of the oviduct in laying hens. [ Method] Immunohistochemical technology and RT-PCR were used to detect the localization of TRPV6 in different reproductive organs. In addition, quantitation of TRPV6 mRNA level and protein concentration in these tissues were carried out by real-time PCR and Western blotting analysis, respectively. [ Result ] Positive expression of the TRPV6 protein and mRNA was observed in ovarium, magnum, isthmus and uterus parts of the oviduct in laying hens, respectively. TRPV6 was localized in the primary follicle and secondary follicle in the ovary. Furthermore, intense staining was observed in villus epithelial cell （face to lumen） of mucous epithelium of the magnum, isthmus and uterus parts of the oviduct. And the weaker staining was also found in the basilar layer of endometrium in the uterus parts of the oviduct. Moreover, the mRNA levels of TRPV6 in the magnum and isthmus were significantly decreased compared with that in the ovary （P〈0.05）. The expression of TRPV6 in the magnum was also lower than that in the ovary （P〈0.05）. However, the expression of TRPV6 mRNA in the uterus parts of the oviduct was lower than that in the ovary, but the change of protein concentration was in contrast, and the difference was not significant （P〉0.05）. [Conclusion] The expression of TRPV6 was detected in ovarium, magnum, isthmus and uterus parts of the oviduct in laying hens. And the higher concentration of TRPV6 in ovary and uterus parts implied that the TRPV6 may be not only involved in the development and maturation, but also may play an important role in Ca^2＋ transport in ESG （eggshell gland） during eggshell calcification.