FERTILIZATION OF THE MOUSE OOCYTE
FERTILIZATION OF THE MOUSE OOCYTE
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DOI：
10.1002/jemt.1060170404
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发表时间：
1991-04-01
期刊：
影响因子：
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通讯作者：
CALARCO, PG
中科院分区：
文献类型：
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作者：
CALARCO, PG
本文介绍了小鼠卵母细胞成熟和受精的形态学数据，回顾了支持精子受体存在的证据，并提出了这一研究领域的未来方向。我们使用扫描电子显微镜在各种条件下检查卵母细胞。无论成熟发生在体内还是体外，成熟小鼠卵母细胞的表面都是相似的。观察到有能力的精子（顶体完整的和顶体反应的）与卵母细胞表面的微绒毛相互作用。对于介导哺乳动物受精的卵母细胞表面蛋白知之甚少。我们和其他人的数据表明，对活的未受精卵进行酶处理会干扰精子结合。酶处理（胰蛋白酶、胰凝乳蛋白酶处理或链霉蛋白酶）减少了结合精子的数量，表明去除了参与受精的表面蛋白。胰蛋白酶处理还会导致中期 II 区域周围带中的表面微绒毛有所延长。代谢标记后，无透明带未受精卵的蛋白质可以通过 SDS-PAGE 和放射自显影来鉴定。未经处理和酶处理的鸡蛋的一维凝胶的比较显示，消化后 263、170、137、97 和 87 kD 的蛋白质几乎完全消失；胰蛋白酶或胰凝乳蛋白酶后 66 kD 蛋白质增加；胰凝乳蛋白酶处理后出现 20 kD 的主要新条带。受精卵显示出 255-265 kD 带的丢失以及其他变化。之前通过表面标记发现了 97 kD 和 87 kD 的蛋白质（Johnson 和 Calarco，1980b），我们的 97 kD 和 66 kD 条带的分子量与 Boldt 等人鉴定的条带相似。 （1989）。总而言之，这些数据确定了一些在卵子表面发挥精子受体作用的候选蛋白质。未来的工作应集中于通过开发受精阻断抗体来鉴定在受精过程中发挥生理功能的表面蛋白质。与其他哺乳动物精子受体的相关性以及所涉及基因的鉴定将为我们理解受精以及不孕和避孕问题提供有价值的信息。
This paper presents morphological data on mouse oocyte maturation and fertilization, reviews evidence supporting the existence of a sperm receptor, and suggests future directions for this line of research. We used scanning electron microscopy to examine oocytes under a variety of conditions. The surfaces of mature mouse oocytes are seen to be similar whether maturation occurs in vivo or in vitro. Capacitated sperm (both acrosome-intact and acrosome-reacted) are observed to interact with the microvilli of the oocyte surface.Little is known about oocyte surface proteins that mediate fertilization in mammals. Data of ours and others show that enzyme treatment of live unfertilized eggs interferes with sperm binding. Enzyme treatment (trypsin, chymotrypsin treatment, or pronase) reduces the number of bound sperm, suggesting removal of a surface protein involved in fertilization. Trypsin treatment also causes some lengthening of surface microvilli in a belt surrounding the metaphase II region. After metabolic labeling, proteins of zona-free unfertilized eggs can be identified by SDS-PAGE and autoradiography. Comparison of 1-D gels from untreated and enzyme-treated eggs show the nearly complete disappearance of proteins of 263, 170, 137, 97, and 87 kD after digestion; an increase in a 66 kD protein after trypsin or chymotrypsin; and a major new band of 20 kD after chymotrypsin treatment. Fertilized eggs show the loss of a 255-265 kD band among other changes. Proteins of 97 kD and 87 kD were seen previously by surface labeling (Johnson and Calarco, 1980b), and our 97 kD and 66 kD bands are similar in molecular weight to those identified by Boldt et al. (1989). Taken together, these data identify a few candidate proteins for the role of sperm receptor on the egg surface.Future work should focus on identification of the surface protein(s) which functions physiologically in fertilization by developing fertilization-blocking antibodies. Relatedness to other mammalian sperm receptors and identification of the genes involved would provide valuable information to our understanding of fertilization and to the problems of infertility and contraception.