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金黄仓鼠发育中胚胎卵裂球内吞小室中输卵管蛋白的免疫定位

Immunolocalization of oviductin in endocytic compartments in the blastomeres of developing embryos in the golden hamster.

作者信息

Kan F W, Roux E, Bleau G

机构信息

Department of Anatomy, Faculty of Medicine, Université de Montréal, Quebec, Canada.

出版信息

Biol Reprod. 1993 Jan;48(1):77-88. doi: 10.1095/biolreprod48.1.77.

Abstract

We have previously localized an antigen of oviductal origin in the zona pellucida of superovulated hamster ova. This antigen is a high-molecular-weight glycoprotein that is secreted by the nonciliated secretory cells of the oviduct and later is transferred to the zona pellucida of the oocyte during oviductal transit. This glycoprotein is rich in N-acetyl-D-galactosamine residues and has been designated Hamster Oviductin-1. In the present study, we have examined the intracellular localization of the oviductin in the blastomeres of developing embryos of the golden hamster. Seventeen cycling females were used for the localization and detection of the oviductin in oviductal oocytes and early embryos. Thin sections of hamster 1-cell oocytes, and 2-cell, and 8-cell embryos, embedded in Lowicryl and then incubated with the monoclonal antibody against the oviductin, revealed a homogenous distribution of antigenic sites in the matrix of the zona pellucida. While immunogold labeling was completely absent in the cytoplasm of 1-cell ova prior to fertilization, labeling was found associated with coated pits and coated vesicles and with flocculent material in the perivitelline space of fertilized eggs. In addition to these labeled endocytic structures, many endosomes, multivesicular bodies, and secondary lysosomes were also found to be labeled heavily in the cytoplasm of developing embryos. Our results indicate that following the transfer of Hamster Oviductin-1 to the zona pellucida of oocytes during transit in the oviduct and subsequent to fertilization, some of the oviductin associated with the zona pellucida appears to be internalized by blastomeres of the embryo and further processed through the endosomal/lysosomal pathway.

摘要

我们之前已将一种输卵管来源的抗原定位在超排仓鼠卵子的透明带中。这种抗原是一种高分子量糖蛋白,由输卵管的非纤毛分泌细胞分泌,随后在卵子通过输卵管的过程中转移到卵母细胞的透明带。这种糖蛋白富含N-乙酰-D-半乳糖胺残基,已被命名为仓鼠输卵管蛋白-1。在本研究中,我们检测了金黄仓鼠发育中胚胎卵裂球内输卵管蛋白的细胞内定位。17只处于发情周期的雌性仓鼠用于输卵管卵母细胞和早期胚胎中输卵管蛋白的定位和检测。将仓鼠1细胞卵母细胞、2细胞和8细胞胚胎制成超薄切片,包埋在Lowicryl中,然后用抗输卵管蛋白的单克隆抗体孵育,结果显示透明带基质中抗原位点分布均匀。在受精前的1细胞卵子细胞质中完全没有免疫金标记,但在受精卵的卵周隙中发现标记与被膜小窝、被膜小泡以及絮状物质相关。除了这些标记的内吞结构外,在发育中胚胎的细胞质中还发现许多内体、多囊泡体和次级溶酶体也被大量标记。我们的结果表明,在仓鼠输卵管蛋白-1在输卵管中运输过程中转移到卵母细胞透明带后,以及受精后,一些与透明带相关的输卵管蛋白似乎被胚胎的卵裂球内化,并通过内体/溶酶体途径进一步加工。

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