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Loss of reactivity to pan-cadherin antibody in epidermal cells as a marker for metamorphic alteration of Xenopus skin

机译:表皮细胞中泛钙黏着蛋白抗体的反应性丧失,这是爪蟾皮肤变态性改变的标志

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Pan-cadherin antibodies recognize the conserved C-terminal region of the family of cell-cell adhesion molecules, cadherins, and have a broad spectrum of reactivity to the molecules. In the present study, by immuno-histochemistry using an anti-pan cadherin monoclonal antibody (mAb), expression dynamics of cadherins in epidermal tissues were analyzed during metamorphosis of Xenopus laevis. At early stages of development, the anti-pan cadherin mAb detected signals at cell-cell boundaries and in the cytoplasm of both trunk and tail epidermal cells. During metamorphosis, the immunoreactivity decreased in the trunk skin tissue but remained in the tail. At the climax stage, immunoreactivity was observed only in the regressing tail epidermis. The signalsdisappeared completely from the trunk epidermis, which had already transformed into adult-type tissue. This observation was confirmed by western blot analysis. A specific band was detected in the larval skin, but not in the adult lysate, at approximately135kDa in molecular size, corresponding to the molecular mass of cadherins. This different immunoreactivity in larvae and adults was observed in the epidermis of the skin, but not in any other tissues examined, that is, brain, kidney and liver. The immunoreactivity seen in larval epidermal cells was drastically downregulated by thyroid hormone treatment in vitro. These changes of immunoreactivity were specific for the C-terminal region of cadherins, suggesting intracellular alteration of the moleculesduring metamorphosis, and the anti-pan cadherin mAb can be a marker for larval-type epidermal cells that is applicable to analysis of Xenopus metamorphosis.
机译:泛钙粘蛋白抗体可识别细胞-细胞粘附分子钙粘蛋白家族的保守C端区域,并具有与分子的广泛反应性。在本研究中,通过使用抗泛钙黏着蛋白单克隆抗体(mAb)的免疫组织化学,分析了非洲爪蟾变态过程中表皮组织中钙黏着蛋白的表达动态。在开发的早期阶段,抗泛钙粘蛋白单克隆抗体在干细胞和尾部表皮细胞的细胞-细胞边界以及细胞质中检测到信号。在变态过程中,躯干皮肤组织中的免疫反应性降低,但保留在尾巴中。在高潮阶段,仅在退化的尾巴表皮中观察到免疫反应性。信号完全从已经转变为成人型组织的躯干表皮中消失。蛋白质印迹分析证实了该观察结果。在幼虫皮肤中检测到一条特定的条带,但在成人裂解物中未检测到一条条带,其分子大小约为135kDa,与钙粘蛋白的分子量相对应。在皮肤的表皮中观察到了幼虫和成虫的这种不同的免疫反应性,但在检查的任何其他组织(即脑,肾和肝)中均未观察到。通过体外甲状腺激素治疗,幼虫表皮细胞中看到的免疫反应性显着下调。这些免疫反应性变化对钙黏着蛋白的C末端区域具有特异性,表明在变态过程中分子的细胞内改变,而抗泛钙黏着蛋白mAb可以作为幼虫型表皮细胞的标志物,可用于分析非洲爪蟾变态。

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