整合素和整合素在猪受精过程中作为卵母细胞粘附分子的潜在作用。

J. Linfor, T. Berger
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引用次数: 45

摘要

整合素分子是一种细胞粘附分子,被认为与啮齿类动物和人类的精子-卵细胞相互作用有关。本研究的目的是评估整合素分子是否存在于猪卵母细胞表面,与该物种参与精子-卵母细胞相互作用一致。采用免疫细胞化学和共聚焦显微镜检测猪卵母细胞质膜上β 1、α 1、α 2、α 3、α 4、α 5、α 6和α v整合素亚基的存在。猪卵母细胞表面一致存在β 1和α v整合素亚基;然而,剩余的α整合素亚基没有被常规检测到。针对β 1和α v整合素亚基的抗体在部分纯化的卵母细胞质膜上识别适当大小的蛋白带。这两种抗体也能识别从精子细胞膜亲和柱中分离的卵母细胞细胞膜蛋白。western sandwich blot显示,137和93 kDa的精子质膜蛋白可能是β 1整合素亚基的配体。β a1整合素亚基胞外结构域的抗体降低了猪精卵细胞结合(P < 0.05),也表明β a1整合素亚基在猪精卵相互作用中起辅助作用。这些结果与猪卵母细胞整合素在受精过程中与精子质膜上的配体相互作用是一致的。
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Potential role of alphav and beta1 integrins as oocyte adhesion molecules during fertilization in pigs.
Integrin molecules are cell adhesion molecules that are thought to be involved in sperm-oocyte interaction in rodents and humans. The objective of this study was to evaluate whether integrin molecules were present on the surface of pig oocytes, consistent with involvement in sperm-oocyte interaction in this species. Immunocytochemistry and confocal microscopy were used to evaluate the presence of beta1, and alpha1, alpha2, alpha3, alpha4, alpha5, alpha6 and alphav integrin subunits on the plasma membrane of pig oocytes. The beta1 and alphav integrin subunits were present consistently at the surface of pig oocytes; however, the remaining alpha integrin subunits evaluated were not routinely detected. The antibodies to the beta1 and alphav integrin subunits recognized appropriately sized protein bands on western blots of partially purified oocyte plasma membrane. These two antibodies also recognized oocyte plasma membrane protein isolated from a sperm plasma membrane affinity column. Sperm plasma membrane proteins of 137 and 93 kDa appeared to be the ligands for the beta1 integrin subunit as revealed by a western sandwich blot. Antibody to an extracellular domain of the beta1 integrin subunit reduced pig sperm-oocyte binding (P < 0.05), also indicating an assisting role for a beta1 oocyte integrin subunit in sperm-oocyte interaction in pigs. These results are consistent with an alphavbeta1 pig oocyte integrin interacting with a ligand on the sperm plasma membrane during fertilization.
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