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大鼠壁层卵黄囊基底膜中纤连蛋白和层粘连蛋白的沉积:免疫组织化学和生物合成研究

Deposition of fibronectin and laminin in the basement membrane of the rat parietal yolk sac: immunohistochemical and biosynthetic studies.

作者信息

Amenta P S, Clark C C, Martinez-Hernandez A

出版信息

J Cell Biol. 1983 Jan;96(1):104-11. doi: 10.1083/jcb.96.1.104.

Abstract

Rat parietal yolk sacs (PYS) at gestational ages 7.5, 9.5, 11.5, 13.5, 14.5, and 16.5 d were reacted with antibodies against laminin or plasma fibronectin. At all times studied, laminin consistently gave a positive reaction with Reichert's membrane and with the cytoplasm of PYS cells. In contrast, fibronectin gave a negative reaction with Reichert's membrane at day 7.5, was weakly positive at day 9.5, and from then on was increasingly positive with maximum reactivity at 14.5 d. By electron microscopic immunohistochemistry, antilaminin reacted strongly with 14.5-d Reichert's membrane and with the contents of the rough endoplasmic reticulum RER cisternae of the PYS cells. Antifibronectin had some spotty reactivity with Reichert's membrane, but the cytoplasm of the PYS cells was negative. The contents of the vitelline vessels and the interface between trophoblast and Reichert's membrane were strongly positive. Metabolic labeling of PYS cells in organ culture clearly demonstrated the presence of laminin, type IV procollagen, and entactin both in the medium and in tissues, but fibronectin was absent. No component in the medium bound to gelatin-Sepharose columns. These studies demonstrate that PYS cells, which actively synthesize and secrete basement membrane components, do not synthesize any detectable fibronectin. Furthermore, the anti-fibronectin staining pattern in the vitelline vessels and trophoblast-Reichert's membrane interface strongly suggests that the fibronectin present in Reichert's membrane is derived from the maternal circulation and is merely "trapped" in the membrane.

摘要

将妊娠7.5、9.5、11.5、13.5、14.5和16.5天的大鼠顶叶卵黄囊(PYS)与抗层粘连蛋白或血浆纤连蛋白的抗体反应。在所有研究的时间点,层粘连蛋白始终与赖歇特氏膜和PYS细胞的细胞质产生阳性反应。相比之下,纤连蛋白在7.5天时与赖歇特氏膜呈阴性反应,在9.5天时呈弱阳性,从那时起呈逐渐阳性,在14.5天时反应性最强。通过电子显微镜免疫组织化学,抗层粘连蛋白与14.5天的赖歇特氏膜以及PYS细胞粗面内质网(RER)池的内容物强烈反应。抗纤连蛋白与赖歇特氏膜有一些斑点状反应,但PYS细胞的细胞质呈阴性。卵黄血管的内容物以及滋养层与赖歇特氏膜之间的界面呈强阳性。器官培养中PYS细胞的代谢标记清楚地表明,培养基和组织中均存在层粘连蛋白、IV型前胶原和巢蛋白,但不存在纤连蛋白。培养基中没有任何成分与明胶-琼脂糖柱结合。这些研究表明,积极合成和分泌基底膜成分的PYS细胞不合成任何可检测到的纤连蛋白。此外,卵黄血管和滋养层-赖歇特氏膜界面中的抗纤连蛋白染色模式强烈表明,赖歇特氏膜中存在的纤连蛋白源自母体循环,只是“被困”在膜中。

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