Verhage H G, Fazleabas A T
Department of Obstetrics and Gynecology, University of Illinois College of Medicine, Chicago 60680.
Endocrinology. 1988 Jul;123(1):552-8. doi: 10.1210/endo-123-1-552.
This study was undertaken to determine the effect of estradiol and progesterone on the synthesis of oviduct-specific proteins and to determine if there were changes in the synthesis of these proteins in different regions of the oviduct. Ovariectomized females were either untreated, treated with estradiol for 7 or 14 days, or primed with estradiol for 14 days and then treated with estradiol plus progesterone or progesterone alone for either 7 or 14 days. Oviducts were incubated in the presence of labeled methionine or glucosamine for 24 h at 37 C, and the culture medium was then analyzed by one- and two-dimensional (2-D) polyacrylamide gel electrophoresis, followed by fluorography. A family of macromolecules [mol wt (Mr), 100,000-130,000] was only present in the animals treated with estradiol for 7 or 14 days. 2-D analysis demonstrated that two proteins, one basic and one acidic, were the major estradiol-responsive proteins in the 100,000-130,000 Mr region. In addition, an acidic protein in this region increased in intensity with estradiol treatment. All three proteins incorporated methionine and glucosamine. Since a steroid-responsive gradient is known to exist in the oviduct, oviducts were divided into fimbria, ampulla, and isthmic regions and cultured separately. 2-D analysis of the medium indicated that the ampulla synthesized both the acidic (Mr, 100,000-120,000) and basic (Mr, 120,000-130,000) proteins, whereas the acidic protein was dominant in the fimbria and the basic protein was dominant in the isthmus. Another acidic protein (Mr, 110,000) was only present in the fimbria and ampulla. This study clearly demonstrates that the estradiol-treated baboon oviduct synthesizes several proteins that were not detectable in the ovariectomized or progesterone-treated animal. Also, the different regions of the oviduct have different synthetic capabilities for these proteins. The electrophoretic characteristics of these proteins are similar to those we have observed at midcycle in the intact baboon and human oviduct.
本研究旨在确定雌二醇和孕酮对输卵管特异性蛋白合成的影响,并确定输卵管不同区域这些蛋白的合成是否存在变化。对去卵巢的雌性动物,要么不进行处理,要么用雌二醇处理7天或14天,要么先用雌二醇预处理14天,然后再用雌二醇加孕酮或单独用孕酮处理7天或14天。将输卵管在含有标记甲硫氨酸或氨基葡萄糖的条件下于37℃孵育24小时,然后通过一维和二维(2-D)聚丙烯酰胺凝胶电泳分析培养基,随后进行荧光显影。一类大分子物质[分子量(Mr),100,000 - 130,000]仅在接受雌二醇处理7天或14天的动物中存在。二维分析表明,在100,000 - 130,000 Mr区域,两种蛋白,一种碱性蛋白和一种酸性蛋白,是主要的雌二醇反应性蛋白。此外,该区域的一种酸性蛋白在雌二醇处理后强度增加。所有这三种蛋白都掺入了甲硫氨酸和氨基葡萄糖。由于已知输卵管中存在类固醇反应梯度,因此将输卵管分为伞部、壶腹部和峡部区域并分别培养。对培养基的二维分析表明,壶腹部合成了酸性(Mr,100,000 - 120,000)和碱性(Mr,120,000 - 130,000)蛋白,而酸性蛋白在伞部占主导,碱性蛋白在峡部占主导。另一种酸性蛋白(Mr,110,000)仅存在于伞部和壶腹部。本研究清楚地表明,经雌二醇处理的狒狒输卵管合成了几种在去卵巢或经孕酮处理的动物中无法检测到的蛋白。而且,输卵管的不同区域对这些蛋白具有不同的合成能力。这些蛋白的电泳特征与我们在完整狒狒和人类输卵管月经周期中期观察到的相似。