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猪和兔卵巢早期卵泡发育过程中层粘连蛋白的定位

Localization of laminin proteins during early follicular development in pig and rabbit ovaries.

作者信息

Lee V H, Britt J H, Dunbar B S

机构信息

Department of Cell Biology and Biochemistry, Texas Tech University, Lubbock 79430, USA.

出版信息

J Reprod Fertil. 1996 Sep;108(1):115-22. doi: 10.1530/jrf.0.1080115.

Abstract

Laminin, a major component of the basal lamina, is known to be important in the differentiation of epithelial cells. The outer granulosa cell layer of ovarian follicles is attached to a basal lamina surrounding the follicle and it has been demonstrated that proteins of the basal lamina can alter the steroidogenic capacity and cytoskeletal composition of mature granulosa cells. The present studies were carried out to evaluate the developmental expression of laminin proteins in the basal lamina of ovarian follicles during early stages of granulosa cell differentiation. Ovaries from sexually immature pigs (0-20 weeks of age) and rabbits (0-12 weeks of age) were used for this study because formation of primordial follicles and initiation of follicular growth occur partially or completely postnatally. Specific antibodies were made against laminin beta 1/gamma 1 chains using protein purified by high resolution two-dimensional polyacrylmide gel electrophoresis. These antibodies were used in immunohistochemical localization studies to demonstrate that laminin is associated with the basal lamina surrounding egg clusters and their connections to the ovarian surface epithelium and ovarian rete during early ovarian development. In addition, laminin was observed to associate with a continuous matrix that surrounds forming primordial follicles as they are isolated from the egg clusters. Laminin is localized in the basal lamina of primordial, primary, secondary, and tertiary follicles of both pig and rabbit ovaries. Laminin proteins are not only expressed throughout early follicular development in pigs and rabbits, but are also found surrounding the germ cell compartments prior to initiation of meiosis and formation of primordial follicles. These results demonstrate that laminin proteins are deposited in the basal lamina well before granulosa cells undergo morphological differentiation. This pattern of expression suggests that laminin proteins alone do not control changes in granulosa cell morphology during early development but may be required to maintain cell lineage commitment.

摘要

层粘连蛋白是基膜的主要成分,已知其在上皮细胞分化中起重要作用。卵巢卵泡的外层颗粒细胞层附着于卵泡周围的基膜,并且已经证明基膜蛋白可以改变成熟颗粒细胞的类固醇生成能力和细胞骨架组成。本研究旨在评估颗粒细胞分化早期卵巢卵泡基膜中层粘连蛋白的发育表达情况。本研究使用了性未成熟猪(0 - 20周龄)和兔(0 - 12周龄)的卵巢,因为原始卵泡的形成和卵泡生长的起始部分或全部发生在出生后。使用通过高分辨率二维聚丙烯酰胺凝胶电泳纯化的蛋白质制备针对层粘连蛋白β1/γ1链的特异性抗体。这些抗体用于免疫组织化学定位研究,以证明层粘连蛋白在卵巢早期发育过程中与卵团周围的基膜及其与卵巢表面上皮和卵巢网的连接有关。此外,观察到层粘连蛋白与围绕从卵团中分离出来的正在形成的原始卵泡的连续基质相关联。层粘连蛋白定位于猪和兔卵巢的原始卵泡、初级卵泡、次级卵泡和三级卵泡的基膜中。层粘连蛋白不仅在猪和兔的早期卵泡发育过程中均有表达,而且在减数分裂开始和原始卵泡形成之前的生殖细胞区室周围也能发现。这些结果表明,层粘连蛋白在颗粒细胞发生形态分化之前就已沉积在基膜中。这种表达模式表明,仅层粘连蛋白本身并不能控制早期发育过程中颗粒细胞形态的变化,但可能是维持细胞谱系定向所必需的。

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