Secretion and Fibril Formation of Vitelline Membrane from Avian Granulosa cells

禽颗粒细胞卵黄膜的分泌和原纤维形成

• 批准号: 15380191
• 负责人: MORI Makoto
• 金额: $ 10.05万
• 依托单位: Shizuoka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15380191/
• 关键词: Vitelline membrane; Oocyte; Quail; Domestic fowl; Egg coat; Gene; Extracellular matrix; Aves; ZPC; ZP1; 卵巣; タンパク分泌; 繊維形成; 透明帯; 卵胞; 肝臓; CHO細胞

1.The extracellular matrix surrounding the oocyte before ovulation is called perivitelline membrane in avian species. The perivitelline membrane is constructed with two major glycoproteins, ZPC and ZP1 that are synthesized in the ovarian granulosa cells and the liver, respectively. Although the properties of the major components of the perivitelline membrane had been examined, the knowledge of the nature of fibril formation and secretion pattern from the cell is lacking. In this study, we focused on the binding of quail ZPC and quail ZP1, and comparison of gene structure of various avian species. Moreover, the hypothesis that ZPC binding protein shows an important role for the fibril formation in immature follicle was examined.2.The microsomal fraction of the granulosa cell was immunoblotted with anti-ZPC after SDS-polyacrylamido gel electrophoresis, and a novel protein migrated at 200 kDa was detected. This band was not reacted with anti-ZPC antiserum or anti-ZP1 antiserum, but strongly bind to ZPC. These results indicated the existence of ZPC binding protein for the fibril formation of perivitelline membrane.3.In order to compare the binding affinity of quail ZPC to ZP1 of the various avian species, the gel electrophoresis analysis was performed and found that quail ZPC can bind to other partidge ZP1. Turkey and duck ZP1 was cloned their nucleic acid sequences were determined. The results showed that the protein has variance at C terminal amino acids. Moreover, mutant ZP1 was expressed in CHO-K1 cells, and it is bind to ZPC strongly. These results indicated that the N-terminal proteolytic processing of ZP1 might take place after the arrival of ZP1 at the ovary, and the resulting product is incorporated into the perivitelline membrane in species-specific manner.

1.在鸟类中，排卵前卵母细胞周围的细胞外基质称为卵周膜。卵周膜由两种主要的糖蛋白ZPC和ZP 1构成，它们分别在卵巢颗粒细胞和肝脏中合成。尽管已经检查了卵黄周膜主要成分的性质，但对细胞原纤维形成和分泌模式的本质还缺乏了解。在本研究中，我们重点研究了鹌鹑ZPC和鹌鹑ZP 1的结合，并比较了不同鸟类的基因结构。2.颗粒细胞微粒体部分经SDS-聚丙烯酰胺凝胶电泳后，用抗ZPC抗体进行免疫印迹，发现一个分子量为200 kDa的新蛋白。该条带与抗ZPC抗血清或抗ZP 1抗血清不反应，但与ZPC强结合。这些结果表明，ZPC结合蛋白的存在促进了卵周膜纤维的形成。3.为了比较鹌鹑ZPC与不同鸟类ZP 1的结合亲和力，通过凝胶电泳分析，发现鹌鹑ZPC可以与ZP 1的其它部分结合。克隆了火鸡和鸭的ZP 1基因，并测定了其核酸序列。结果表明，该蛋白质在C端氨基酸上存在变异。此外，突变体ZP 1在CHO-K1细胞中表达，并与ZPC强烈结合。这些结果表明，ZP 1的N-末端蛋白水解加工可能发生在ZP 1到达卵巢后，所产生的产物以种特异性的方式掺入到卵黄周膜中。

项目成果

Accumulation of ZP1 and ZPC in Quail Perivitelline Membrane During Follicular Development

卵泡发育过程中鹌鹑卵周膜中 ZP1 和 ZPC 的积累

• 发表时间: 2004
• 期刊: Journal of Poultry Science 41(4)
• 作者: Mariko Mochizuki;Makoto Mori;Mayumi Akinaga;Kyoko Yugami;Chika Oya;Ryo Hondo;Fukiko Ueda;Ahmed M. Hanafy;Tomohiro Sasanami;Mariko Mochizuki;Ahmed M.Hanafy;Tomohiro Sasanami
• 通讯作者: Tomohiro Sasanami

Jianzhi Pan: "Stimulation of ZPC Production by Follicle-Stimulating Hormone in the Granulosa Cells of Japanese Quail (Coturnix japonica)"Journal of Poultry Science. 40(3). 202-211 (2003)

潘建志：“日本鹌鹑（Coturnix japonica）颗粒细胞中卵泡刺激素对 ZPC 产生的刺激”家禽科学杂志。

Tomohiro Sasanami: "Asparagine-Linked Oligosaccaride-Independent Secretion of Egg Envelope Glycoprotein ZPC of the Japanese Quail (Coturnix japonica)."Comparative Biochemistry and Physiology. 134(3). 631-638 (2003)

Tomohiro Sasanami：“日本鹌鹑（Coturnix japonica）卵膜糖蛋白 ZPC 的天冬酰胺连接寡糖独立分泌。”比较生物化学和生理学。

Binding of Xenoestrogens and Phytoestrogens to Estrogen Receptor β of Japanese Quail (Coturnix japonica)

异种雌激素和植物雌激素与日本鹌鹑 (Coturnix japonica) 雌激素受体 β 的结合

• 发表时间: 2005
• 期刊: Journal of Poultry Science 42
• 作者: Ahmed M.Hanafy

A Novel Assay System of Chemicals Using In-Vitro Maturation of Mouse Oocytes

利用小鼠卵母细胞体外成熟的新型化学物质检测系统

• 发表时间: 2004
• 期刊: Journal of Mammalian Ova Research 21(3)
• 作者: Mariko Mochizuki;Makoto Mori;Mayumi Akinaga;Kyoko Yugami;Chika Oya;Ryo Hondo;Fukiko Ueda;Ahmed M. Hanafy;Tomohiro Sasanami;Mariko Mochizuki;Ahmed M.Hanafy;Tomohiro Sasanami;Ryota Tanaka
• 通讯作者: Ryota Tanaka