摘要
为在胚胎共培养过程中添加相关激素提高哺乳动物胚胎发育的研究提供理论依据,本研究探讨了促卵泡生成素(FSH)对牦牛输卵管上皮细胞分泌特异性糖蛋白的影响。体外分离培养牦牛输卵管上皮细胞,并在细胞中添加不同浓度的FSH,作用6 h后运用荧光定量PCR分析输卵管特异性糖蛋白mRNA的表达水平,并用细胞免疫标记对其分泌输卵管蛋白的部位进行分析。结果显示,FSH的浓度为0.5-5.0μg/m L时,输卵管蛋白的表达量随着FSH浓度的上升而增加,在5.0μg/m L的FSH作用后,输卵管蛋白的mRNA表达量最高;浓度超过10.0μg/m L时,输卵管蛋白基因的表达量降低。结果表明,FSH具有促进输卵管上皮细胞分泌输卵管特异性糖蛋白的作用,且具有剂量依赖性,其最佳作用浓度为5.0μg/m L,输卵管蛋白主要由细胞质分泌。
To examine the effects of gonadotropin on the development of embryos this study investigated the effect of Follicle Stimulating Hormone（ FSH） on the glycoprotein produced by oviduct epithelial cells. Cultured bovine oviduct epithelial cells were treated for 6 hours with different doses of FSH,and oviductin expression was analyzed by real time quantitative polymerase chain reaction（ RT- q PCR） and immunohistochemistry. Results showed the effective range of FSH（ 0. 5-5. 0 μg / m L）,and quantitative PCR showed the highest oviductin mRNA copy numbers at 5. 0 μg / m L,and reduced mRNA expression during all others. However mRNA expression was decreased when FSH concentration over 10. 0 μg / m L. So,FSH can promote the secretion of oviductin in bovine oviduct epithelial cells at this range of concentrations and 5. 0 μg / m L is the optimal concentration,and it showed a dose-dependent effect. Meanwhile,oviduct protein was secreted in cytoplasm.
出处
《兽类学报》
CAS
CSCD
北大核心
2015年第2期190-195,共6页
Acta Theriologica Sinica
基金
国家自然科学基金资助项目(31272616)
甘肃省科技支撑计划项目-农业类1204NKCA075
