Immunolocalization of the sea urchin sperm receptor in eggs and maturing ovaries.
Immunolocalization of the sea urchin sperm receptor in eggs and maturing ovaries.
复制标题
海胆精子受体在卵子和成熟卵巢中的免疫定位。
DOI：
10.1095/biolreprod41.2.323
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发表时间：
1989
影响因子：
3.6
通讯作者：
Lennarz,WJ
中科院分区：
文献类型：
--
作者：
Ruiz-Bravo,N;Janak,DJ;Lennarz,WJ
紫球海胆卵的精子受体是一种高分子量蛋白多糖样分子，在竞争生物测定中具有种属特异性的受精抑制作用。用含有一个主要N端序列的高活性精子受体制备兔抗精子抗体。该抗体在低浓度下特异性抑制受精，而不干扰受精膜的升高。在免疫荧光显微镜下，抗体识别成熟卵细胞表面和表面下的皮质颗粒中的决定簇。预吸收的抗体与钙可溶性部分的渗出液从皮质颗粒呈现的抗体特异性的细胞表面在成熟的鸡蛋，仍然能够抑制受精在相同的浓度前处理皮质颗粒渗出液。用皮质颗粒预吸附抗体，电镜下计数抗体-金颗粒，发现在季节前卵巢未成熟卵母细胞的细胞表面几乎检测不到精子受体，在产卵季节开始或高峰期采集的卵巢未成熟卵母细胞的细胞表面和细胞内有精子受体，而在卵巢腔中成熟卵母细胞的细胞表面仅存在精子受体。我们的研究结果表明，受体在卵子发生早期合成，并迅速移动到卵细胞表面。
The sperm receptor from Strongylocentrotus purpuratus eggs is a high molecular weight proteoglycan-like molecule that inhibits fertilization species-specifically in a competition bioassay. A preparation of highly active sperm receptor that contained one major N-terminal sequence was used to generate polyclonal antibody in rabbits. This antibody species-specifically inhibited fertilization at low concentrations without interfering with fertilization envelope elevation. On immunofluorescence microscopy, the antibody recognized determinants on the mature egg cell surface and in the cortical granules just beneath the surface. Preabsorption of the antibody with the calcium-soluble fraction of the exudate from cortical granules rendered the antibody specific for the cell surface in mature eggs and still able to inhibit fertilization at the same concentrations as before treatment with cortical granule exudate. With antibody preabsorbed with cortical granule and by counting antibody-gold particles viewed by electron microscopy, sperm receptor was almost undetectable on the cell surface of immature oocytes in preseason ovaries, present on the cell surface and intracellularly in immature oocytes of ovaries collected at the beginning or at the height of the spawning season, and present only on the cell surface of mature oocytes in the lumen of the ovaries. Our results indicate that receptor is synthesized early in oogenesis and is rapidly moved to the egg cell surface.