摘要
本研究对兔输卵管上皮细胞（ＲＯＥＣ）分泌蛋白（ＲＯＰｓ）的特征及功能作了初步研究和分析。ＲＯＲｓ分子量介于１３５—４４ｋＤ，可分为３组，其中第２、３组为酸性糖蛋白。经糖苷酶消化，第３组ＲＯＰｓ显著地降解为分子量接近３０ｋＤ的蛋白。利用制备的抗总ＲＯＰｓ多抗（Ａｒｏｐ－Ｔ）以及针对不同分子量ＲＯＰ的抗血清（Ａｒｏｐ－Ⅰ、Ａｒｏｐ－Ⅱ和Ａｒｏｐ－Ⅲ），分别进行功能分析发现，Ａｒｏｐ－Ｔ和Ａｒｏｐ－Ⅰ能使受精卵发育完全阻断；Ａｒｏｐ－Ⅱ阻断受精卵发育的百分率为７５％；Ａｒｏｐ－Ⅲ无特异性地阻断受精卵发育的作用。结合我们已报道的研究结果，推测ＲＯＰ－６８和ＲＯＰ－５５就是具备启动早期胚胎合子型基因开放、实现正常发育调控过渡功能的输卵管细胞因子。
The development of multicellular animals depends on cell-cell communication, and one of the mediators for the communication is cytokines. After fertilization, the development of mammalian early embryos during transition from maternal to zygotic genetic control requires the stimulation of external signal excreted from oviduct epithelial cells (OEC), otherwise, the developmental block occurs. We studied on the characterization and biological function of rabbit oviductal epithelial cells proteins (ROPs). There are three sets of ROPs with molecular weight of ROPs ranging from 44 kD to 135 kD, and their isoelectric points ranged from 4.5 to 5.9. Two sets of them were determined to be acid glycoproteins by treatment of WGA lectin sepharose and analysis of 2-D SDS-PAGE. After digestion with N- and O-glycosidase, the third set of the glycoproteins were degraded to become two or three spots with molecular weight about 30 kD or less, and their isoelectric points changed to about 5.5 and 5.6. Furthermore, the function of ROPs was analysed by using polyclonal antibody and antiserum against total ROPs (Arop-T) and against different molecular weight of ROP (Arop-Ⅰ, -Ⅱ and -Ⅲ). Arop-T and Arop-Ⅰ respectively completely arrest the development of mouse early embryos, and Arop-Ⅱ blocked the development of 75% mouse embryos, but Arop-Ⅲ almost cannot significantly arrest their preimplantation development. Immunoblotting analysis indicated that Arop-T and Arop-Ⅰ positively reacted on both ROP-68 and ROP-55, and that Arop-Ⅱ only positively reacted on ROP-68. The data conincide with our previous result of 125 Ⅰ-labelling ROPs in the zona pellucida of embryos. These results suggested that ROP-68 and ROP-55 were the oviductal cytokine inducing initiation of zygotic gene and realizing normal transition of maternal to zygotic control of early embryo development.
基金
国家自然科学基金
上海市青年科技启明星计划资助
关键词
输卵管细胞因子
胚胎
发育
兔
Oviductal cytokine, Function analysis, Early embryo, Developmental block
