Verhage H G, Boice M L, Mavrogianis P, Donnelly K, Fazleabas A T
Department of Obstetrics and Gynecology, University of Illinois College of Medicine, Chicago 60680.
Endocrinology. 1989 May;124(5):2464-72. doi: 10.1210/endo-124-5-2464.
Oviducts obtained from estradiol-treated ovariectomized baboons synthesize and release a family of high mol wt (100,000-130,000) glycoproteins during short term explant culture. The objective of this study was to make a polyclonal antibody to these glycoproteins and then use the antibody to determine the presence of the glycoproteins in oviduct flushings, tissue culture media, and tissues obtained from cycling and steroid-treated baboons. Oviduct culture medium proteins from estradiol-treated baboons were separated on one-dimensional polyacrylamide gels and transferred to nitrocellulose membranes. The region containing the glycoproteins was cut out, solubilized in dimethylsulfoxide, mixed with Freund's adjuvant, and injected at 2-week intervals into a male rabbit. The anti-serum used in this study was obtained 6 weeks after the initial injection and cross-reacted with antigens on Western blots of oviduct flushings and oviduct culture media obtained from follicular stage and estradiol-treated baboons. The antigens were absent in oviduct flushings obtained from luteal stage, ovariectomized and estradiol-primed baboons treated with estradiol and progesterone or progesterone alone. The antigens were not detected on Western blots of other reproductive and nonreproductive tract culture media or in serum obtained from follicular stage or estradiol-treated baboons. Immunoperoxidase staining was limited to discrete granules in the apical cytoplasm of secretory cells in oviducts obtained from follicular stage and estradiol-treated baboons. Thus, the secretory cells of the baboon oviduct synthesize and secrete a family of estradiol-dependent oviduct-specific glycoproteins that may have potential functional significance during fertilization and embryo development.
从接受雌二醇处理的去卵巢狒狒身上获取的输卵管，在短期外植体培养过程中会合成并释放一族高分子量（100,000 - 130,000）的糖蛋白。本研究的目的是制备针对这些糖蛋白的多克隆抗体，然后使用该抗体来确定输卵管冲洗液、组织培养基以及从处于发情周期和接受类固醇处理的狒狒身上获取的组织中是否存在这些糖蛋白。来自接受雌二醇处理的狒狒的输卵管培养基蛋白在一维聚丙烯酰胺凝胶上进行分离，然后转移到硝酸纤维素膜上。切下含有糖蛋白的区域，溶解于二甲基亚砜中，与弗氏佐剂混合，并每隔2周注射到一只雄性兔子体内。本研究中使用的抗血清在初次注射6周后获得，并且与从卵泡期和接受雌二醇处理的狒狒的输卵管冲洗液和输卵管培养基的蛋白质免疫印迹上的抗原发生交叉反应。在黄体期、去卵巢并用雌二醇预处理后再用雌二醇和孕酮或仅用孕酮处理的狒狒的输卵管冲洗液中不存在这些抗原。在其他生殖和非生殖道培养基的蛋白质免疫印迹上或从卵泡期或接受雌二醇处理的狒狒获得的血清中未检测到这些抗原。免疫过氧化物酶染色仅限于从卵泡期和接受雌二醇处理的狒狒身上获取的输卵管分泌细胞顶端细胞质中的离散颗粒。因此，狒狒输卵管的分泌细胞合成并分泌一族依赖雌二醇的输卵管特异性糖蛋白，这些糖蛋白在受精和胚胎发育过程中可能具有潜在的功能意义。
