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壁蜥蜴(Podarcis muralis)再生表皮中细胞角蛋白的免疫细胞化学和电泳分布

Immunocytochemical and electrophoretic distribution of cytokeratins in the regenerating epidermis of the lizard Podarcis muralis.

作者信息

Alibardi L, Maurizii M G, Taddei C

机构信息

Dipartimento di Biologia Evoluzionistica Sperimentale, University of Bologna, Bologna, Italy.

出版信息

J Morphol. 2000 Dec;246(3):179-91. doi: 10.1002/1097-4687(200012)246:3<179::AID-JMOR2>3.0.CO;2-D.

Abstract

Using immunocytochemistry at light- and electron-microscope levels, we studied the distribution of three monoclonal antibodies (AE1, AE2, AE3) specific for mammalian alpha-keratins in regenerating lizard epidermis. We also characterized the keratins expressed during this process by immunoblotting after electrophoretic separation. The AE1 antibody is localized in the basal and suprabasal layers of prescaling and scaling epidermis. During the first stages of scale neogenesis, the AE1 antibody also marks the differentiating oberhautchen and beta-layer, but it disappears from these layers as they mature. This antibody does not stain the prekeratinized and keratinized outermost layers in the hinge region. The AE2 antibody labels the superficial wound epidermis, prekeratinizing and keratinized beta- and alpha-layers, but not basal and suprabasal cells. The AE3 antibody labels all living and keratinized epidermal layers, although AE3 immunoreactivity decreases and disappears as the beta-layer matures. The ultrastructural study shows that the AE2 and AE3, but not the AE1, antibodies specifically label small electron-dense areas within the beta-layer, suggesting retention of alpha-keratins. In the stages of tail regeneration examined, immunoblotting with the three antibodies used for the immunolocalization gives a pattern similar to that of the normal epidermis, except distally, where the process of scale differentiation begins. In this region, in addition to the keratin forms discovered in the normal and in proximal regenerating epidermis, an intense low molecular weight band at 40-41 kDa, positive to all three antibodies, is clearly detectable. Furthermore, in the distal region AE1 and AE3 antibodies, but not the AE2, recognize a weak band at 77-78 kDa not present in the normal and proximal epidermis. The localization and the possible role of the different keratins in the regenerating epidermis is discussed.

摘要

我们利用光镜和电镜水平的免疫细胞化学技术,研究了三种对哺乳动物α-角蛋白具有特异性的单克隆抗体(AE1、AE2、AE3)在再生蜥蜴表皮中的分布情况。我们还通过电泳分离后的免疫印迹法,对该过程中表达的角蛋白进行了特征分析。AE1抗体定位于鳞片形成前期和鳞片形成期表皮的基底层和基底上层。在鳞片新生的最初阶段,AE1抗体也标记正在分化的表层和β层,但随着这些层的成熟,它会从这些层中消失。该抗体不会对铰链区的前角质化和角质化最外层进行染色。AE2抗体标记浅表伤口表皮、前角质化和角质化的β层和α层,但不标记基底层和基底上层细胞。AE3抗体标记所有活的和角质化的表皮层,尽管随着β层的成熟,AE3免疫反应性会降低并消失。超微结构研究表明,AE2和AE3抗体(而非AE1抗体)特异性标记β层内的小电子致密区域,提示α-角蛋白的保留。在所检查的尾巴再生阶段,用于免疫定位的三种抗体进行免疫印迹得到的模式与正常表皮相似,但在鳞片分化开始的远端区域除外。在该区域,除了在正常和近端再生表皮中发现的角蛋白形式外,还可清楚检测到一条强烈的40 - 41 kDa低分子量条带,对所有三种抗体均呈阳性。此外,在远端区域,AE1和AE3抗体(而非AE2抗体)识别出一条77 - 78 kDa的弱条带,该条带在正常和近端表皮中不存在。本文讨论了不同角蛋白在再生表皮中的定位及其可能的作用。

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