大阪鲫鱼两种卵黄蛋白免疫细胞化学的研究

THE IMMUNOCYTOCHEMICAL LOCALIZATION OF LIPOVITELLIN AND YOLK PROTEIN L IN CARSSIUS AURATUS CUVIERI

以电泳提纯的卵黄脂磷蛋白和卵黄蛋白Ｌ制备兔抗两种蛋白的抗血清。采用ＰＡＰ法对性腺成熟雌性大阪鲫鱼的肝细胞和卵母细胞进行两种蛋白免疫细胞化学定位研究。肝细胞的粗面内质网上有强烈的卵黄脂磷蛋白的阳性反应，特别是在线粒体的基质中也发现卵黄脂磷蛋白的阳性反应。而另外一种类似于卵黄高磷蛋白的卵黄蛋白──卵黄蛋白Ｌ在肝细胞的粗面内质网和线粒体均呈现阴性反应。提示卵黄脂磷蛋白的前体物质存在于肝细胞的粗面内质网和线粒体中，而卵黄蛋白Ｌ则不是来源于肝细胞。卵母细胞的卵黄颗粒上两种蛋白均呈现阳性反应。卵母细胞外的滤泡细胞也存在卵黄脂磷蛋白的阳性反应。透明带和卵母细胞表面的凹陷中存在卵黄脂磷蛋白的阳性反应颗粒物质，并发现大量纤维状物质存在于透明带和卵母细胞表面。这揭示卵黄脂磷蛋白的前体物质可能首先进入滤泡细胞，分解后经透明带被卵母细胞吞噬摄入，并与卵黄蛋白Ｌ共同存在于卵黄颗粒上。

Lipovitellin and yolk protein L were purified from ovary of Carssius auratus Cuvieri. The rabbit antiserums of these two protein were made. PAP positive reaction of lipovitellin in mitochondrium and endoplasmic reticulum of liver cells was shown. But none of positive reaction of yolk protein L,a protein simillar to phosivitin, was found in liver cells. So we could see that lipovitellin was originated from liver cells (as vitellogenin). But the Yolk protein L was not come from liver cell. There were positive reaction of two proteins in yolk granules of oocyte. It was found that the follicle cells also had positive reaction of lipovitellin. The positive particles and fibillous materials could be seen among microvillus in zona pelucida. This suggested that vitellogenin was first uptaken by follicle cells, and then transported to the surface of oocyte with the help of fibra and entered oocyte through micropinooytosis and formed yolk granules.