Developmental expression of the rabbit 55-kDa zona pellucida protein and messenger RNA in ovarian follicles.
Developmental expression of the rabbit 55-kDa zona pellucida protein and messenger RNA in ovarian follicles.
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兔卵泡中 55-kDa 透明带蛋白和信使 RNA 的发育表达。
DOI：
10.1006/dbio.1993.1036
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发表时间：
1993
影响因子：
2.7
通讯作者：
Dunbar,BS
中科院分区：
文献类型：
--
作者：
Lee,VH;Dunbar,BS
利用免疫学和cDNA探针研究了卵巢卵泡发育过程中与卵母细胞激活和颗粒细胞（GC）分化早期阶段相关的事件。由于透明带（ZP）蛋白在卵巢发育早期是卵母细胞活化的良好标记，因此我们开发了一个分子量为57,185 （rc55）的全长ZP蛋白cDNA，以及一个针对兔ZP蛋白（R55）的特异性抗体。卵巢免疫组织化学分析表明，R55定位于2周及以上动物的卵泡中。R55定位于原始卵泡的卵母细胞和初级卵泡的卵母细胞和GCs中，但在大腔卵泡的GCs中未检测到。从6周龄家兔卵巢中分离GCs，用GCsin体外培养兔ZP蛋白。兔ZP蛋白定位于培养的GCs细胞质中，如果这些细胞被莫能菌素（一种阻断糖蛋白分泌的化合物）处理。cDNA探针rc55的Northern blot分析显示，培养的GCs中含有R55 mRNA，并且R55 mRNA在兔卵巢发育中存在分期特异性表达。免疫印迹分析表明，R55在4周时可检测到，并在6、8和12周的卵巢中积累，这些卵巢含有发育中的原发性和继发性卵泡。总的来说，这些数据说明了R55在兔卵泡卵母细胞和GCs中的发育表达。
The events associated with the early stages of oocyte activation and granulosa cell (GC) differentiation during ovarian follicular development have been investigated using immunological and cDNA probes. Because zona pellucida (ZP) proteins provide excellent markers for oocyte activation during early stages of ovarian development, a cDNA encoding a full-length ZP protein with calculated molecular weight of 57,185 (rc55) and a specific antibody to the corresponding native rabbit ZP protein (R55) were developed. Immunohistochemical analysis of ovaries demonstrated that R55 was localized in follicles of animals 2 weeks and older. R55 was localized in the oocytes of primordial follicles and the oocytes and GCs of primary follicles but was undetectable in the GCs of large antral follicles. GCs were isolated from 6-week-old rabbit ovaries and cultured in defined media to demonstrate expression of rabbit ZP proteins by GCsin vitro. Rabbit ZP proteins were localized in the cytoplasm of cultured GCs if these cells were treated with monensin, a compound that blocks glycoprotein secretion. Northern blot analysis with the cDNA probe rc55 revealed that cultured GCs contained a mRNA for R55 and that there is stage-specific expression of the R55 mRNA in developing rabbit ovaries. Immunoblot analysis demonstrated that R55 is detectable at 4 weeks and accumulates in 6-, 8-, and 12-week ovaries which contain developing primary and secondary follicles. Collectively, these data illustrate developmental expression of R55 in oocytes and GCs of rabbit ovarian follicles.