Institute of Reproductive Immunology, Jinan University, Guangzhou 510632, China.
Cell Biochem Funct. 2010 Dec 2;28(8):661-7. doi: 10.1002/cbf.1704.
In order to elucidate the biogenesis of mouse zona pellucida 2 (mZP2) protein, RT-PCR, and in situ hybridization were carried out to localize the expression of mouse ZP2 mRNA. Cumulus cells of the OCC (Oocyte-Cumulus cell Complex) were isolated from the oocytes after superovulation for the RNA extraction. The frozen sections of ovaries from adolescent and aged mice were prepared to hybridize with RNA probe of mouse ZP2. mRNA of ZP2 was detected in isolated cumulus cells by RT-PCR. Results of in situ hybridization showed that the mRNA of ZP2 was synthesized in both oocyte and granulosa cells at different folliculogenesis stages; and the expression of ZP2 mRNA in granulosa cells was stronger than that in oocyte; much weaker expression of mZP2 was detected in the follicles of aged mouse. These suggest that the entire amount of ZP2 mRNA generated in the granulosa cells layer should be much more than that in oocyte. Therefore, we think that the granulosa cells contribute more to the mZP2 mRNA synthesis than oocyte does.
为了阐明小鼠卵透明带 2（mZP2）蛋白的生物发生，进行了 RT-PCR 和原位杂交，以定位小鼠 ZP2 mRNA 的表达。从超排卵后的卵母细胞中分离出 OCC（卵-卵丘细胞复合物）的卵丘细胞，用于提取 RNA。用小鼠 ZP2 的 RNA 探针制备青春期和老年期小鼠卵巢的冷冻切片进行杂交。通过 RT-PCR 检测到分离的卵丘细胞中 ZP2 mRNA 的存在。原位杂交的结果表明，ZP2mRNA 在不同的卵泡发生阶段的卵母细胞和颗粒细胞中合成；颗粒细胞中 ZP2mRNA 的表达强于卵母细胞；在老年小鼠的卵泡中检测到 mZP2 的表达明显减弱。这表明，颗粒细胞层中产生的 ZP2mRNA 的总量应该比卵母细胞中产生的多得多。因此，我们认为颗粒细胞比卵母细胞对 mZP2mRNA 的合成贡献更大。
