Peng Jiayin, Gao Kexin, Gao Teyang, Lei Yingnan, Han Peng, Xin Haiyun, An Xiaopeng, Cao Binyun
College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, P.R. China.
College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, P.R. China.
Theriogenology. 2015 Dec;84(9):1636-43. doi: 10.1016/j.theriogenology.2015.09.003. Epub 2015 Sep 10.
Tissue inhibitors of metalloproteinases (TIMPs) are associated with several reproductive processes, such as mammalian follicular growth, ovulation, CL formation, and embryonic development. However, the expression and function of TIMPs in goat oviducts remain unclear. This work aimed to identify TIMP1 and TIMP3 expression in the goat oviduct during the estrous cycle via immunohistochemistry, real-time polymerase chain reaction (PCR), and functional studies in cultured goat oviductal epithelial cells. Real-time PCR results demonstrated that TIMP1 and TIMP3 messenger RNAs were expressed in all goat oviductal regions at all stages of the estrous cycle. TIMP1 and TIMP3 proteins were also highly expressed in oviductal epithelial cells with very limited expression in other cell types. Oviductal epithelial cells were treated in vitro with various estradiol concentrations (1-100 nM) for 24 hours. The findings showed that TIMP1 expression increased up to 20 nM but then gradually decreased, whereas no significant effects existed among TIMP3 messenger RNA levels. Time-course studies indicated that estradiol significantly increased TIMP1 expression in a time-dependent manner from 8 hours to 24 hours. By contrast, TIMP3 expression was transiently induced in oviductal epithelial cells at 2 and 4 hours after estradiol treatment. Furthermore, treatment with TIMP1 functionally increased the viability of cultured oviductal epithelial cells. Overall, the results suggested that the differential regulation and function between TIMP1 and TIMP3 might be associated with their unique roles in fertilization and early embryonic development.
金属蛋白酶组织抑制剂（TIMPs）与多种生殖过程相关，如哺乳动物卵泡生长、排卵、黄体形成和胚胎发育。然而，TIMPs在山羊输卵管中的表达和功能仍不清楚。本研究旨在通过免疫组织化学、实时聚合酶链反应（PCR）以及对培养的山羊输卵管上皮细胞进行功能研究，来确定发情周期中山羊输卵管内TIMP1和TIMP3的表达情况。实时PCR结果表明，在发情周期的各个阶段，TIMP1和TIMP3信使核糖核酸在山羊输卵管的所有区域均有表达。TIMP1和TIMP3蛋白在输卵管上皮细胞中也高度表达，而在其他细胞类型中的表达非常有限。将输卵管上皮细胞在体外用不同浓度（1 - 100 nM）的雌二醇处理24小时。结果显示，TIMP1的表达在20 nM时升高，但随后逐渐下降，而TIMP3信使核糖核酸水平之间无显著差异。时间进程研究表明，雌二醇从8小时到24小时以时间依赖性方式显著增加TIMP1的表达。相比之下，雌二醇处理后2小时和4小时，TIMP3的表达在输卵管上皮细胞中被短暂诱导。此外，用TIMP1处理在功能上增加了培养的输卵管上皮细胞的活力。总体而言，结果表明TIMP1和TIMP3之间的差异调节和功能可能与其在受精和早期胚胎发育中的独特作用有关。
