Zona pellucida protein ZP2 is expressed in the oocyte of Japanese quail (Coturnix japonica)
Zona pellucida protein ZP2 is expressed in the oocyte of Japanese quail (Coturnix japonica)
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DOI：
10.1530/rep-09-0222
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发表时间：
2010-02-01
期刊：
影响因子：
3.8
通讯作者：
Sasanami, Tomohiro
中科院分区：
文献类型：
--
作者：
Kinoshita, Mihoko;Rodler, Daniela;Sasanami, Tomohiro
禽类卵周层（PL）是与哺乳动物卵母细胞透明膜（ZP）同源的一层膜，至少由三种糖蛋白组成。我们以前的研究表明，基质的成分，ZP 3和ZPD，在卵巢颗粒细胞合成。另一种成分ZP 1在肝脏中合成，并通过血液循环转运到卵巢。在这项研究中，我们报告的cDNA编码鹌鹑ZP 2的分离和表达的雌性鸟类。通过RNA酶保护试验和原位杂交，我们证明ZP 2转录物仅限于小白色卵泡（FSH）的卵母细胞。ZP 2在卵泡发育过程中表达水平显著下降，在卵母细胞中的表达量最高，Western blot和免疫组化分析表明，80 kDa的ZP 2蛋白是真实的ZP 2蛋白，并且ZP 2蛋白在卵母细胞中也有表达。此外，超微结构分析表明，免疫反应ZP 2定位于放射状，卵黄周隙，卵母细胞的胞质中的卵母细胞。通过免疫印迹分析和免疫荧光显微镜，我们检测到的重组ZP 2与ZP 3的可能的相互作用，这种相互作用可能会导致细胞表面的无定形结构的形成。这些结果首次证明了禽ZP基因在卵母细胞中表达，并且卵母细胞中的ZP 2蛋白可能对卵巢未成熟卵泡中PL的形成起作用。生殖（2010年）139 359-371
The avian perivitelline layer (PL), a vestment homologous to the zona pellucida (ZP) of mammalian oocytes, is composed of at least three glycoproteins. Our previous studies have demonstrated that the matrix's components, ZP3 and ZPD, are synthesized in ovarian granulosa cells. Another component, ZP1, is synthesized in the liver and is transported to the ovary by blood circulation. In this study, we report the isolation of cDNA encoding quail ZP2 and its expression in the female bird. By RNase protection assay and in situ hybridization, we demonstrate that ZP2 transcripts are restricted to the oocytes of small white follicles (SWF). The expression level of ZP2 decreased dramatically during follicular development, and the highest expression was observed in the SWF Western blot and immunohistochemical analyses using the specific antibody against ZP2 indicate that the 80 kDa protein is the authentic ZP2, and the immunoreactive ZP2 protein is also present in the oocytes. Moreover, ultrastructural analysis demonstrated that the immunoreactive ZP2 localizes to the zona radiata, the perivitelline space, and the oocyte cytoplasm in the SWF. By means of western blot analysis and immunofluorescence microscopy, we detected a possible interaction of the recombinant ZP2 with ZP3 and that this interaction might lead to the formation of amorphous structure on the cell surface. These results demonstrate for the first time that the avian ZP gene is expressed in the oocyte, and that the ZP2 protein in the oocyte might play a role for the PL formation in the immature follicles of the ovary. Reproduction (2010) 139 359-371