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通过光学显微镜和电子显微镜对大鼠小肠上皮细胞中肌动蛋白进行免疫细胞化学定位。

Immunocytochemical localization of actin in epithelial cells of rat small intestine by light and electron microscopy.

作者信息

Hagen S J, Trier J S

机构信息

Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts 02115.

出版信息

J Histochem Cytochem. 1988 Jul;36(7):717-27. doi: 10.1177/36.7.3290330.

Abstract

We used post-embedding immunocytochemical techniques and affinity-purified anti-actin antibody to evaluate localization of actin in epithelial cells of small intestine by fluorescence and electron microscopy. Small intestine was fixed with 2% formaldehyde-0.1% glutaraldehyde and embedded in Lowicryl K4M. One-micron or thin sections were stained with antibody followed by rhodamine- or colloidal gold-labeled goat anti-rabbit IgG, respectively. Label was present overlying microvilli, the apical terminal web, and the cytoplasm directly adjacent to occluding and intermediate junctions. Label was associated with outer mitochondrial membranes of all cells and the supranuclear Golgi region of goblet cells. Lateral cytoplasmic interdigitations between mature cells and subplasmalemmal filaments next to intrusive cells were densely labeled. The cytoplasm adjacent to unplicated domains of lateral membrane was focally labeled. Label was prominent over organized filament bundles within the subplasmalemmal web at the base of mature cells, whereas there was focal labeling of the cytoplasm adjacent to the basal membrane of undifferentiated cells. Basolateral epithelial cell processes were labeled. Label was focally present overlying the cellular ground substance. Our results demonstrate that actin is distributed in a distinctive fashion within intestinal epithelial cells. This distribution suggests that in addition to its function as a structural protein, actin may participate in regulation of epithelial tight junction permeability, in motile processes including migration of cells from the crypt to the villus tip, in accommodation of intrusive intraepithelial cells and in adhesion of cells to one another and to their substratum.

摘要

我们采用包埋后免疫细胞化学技术和亲和纯化的抗肌动蛋白抗体,通过荧光显微镜和电子显微镜评估小肠上皮细胞中肌动蛋白的定位。小肠用2%甲醛-0.1%戊二醛固定,包埋于Lowicryl K4M中。1微米切片或超薄切片分别用抗体染色,然后用罗丹明或胶体金标记的山羊抗兔IgG染色。标记物出现在微绒毛、顶端终末网以及紧邻封闭连接和中间连接的细胞质上。标记物与所有细胞的线粒体外膜以及杯状细胞的核上高尔基体区域相关。成熟细胞之间的侧向细胞质交错和侵入细胞旁的质膜下细丝被密集标记。紧邻侧膜未折叠区域的细胞质有局灶性标记。标记物在成熟细胞底部质膜下网内有组织的细丝束上很突出,而未分化细胞基底膜相邻的细胞质有局灶性标记。基底外侧上皮细胞突起被标记。标记物局灶性地出现在细胞基质上。我们的结果表明,肌动蛋白在肠上皮细胞内以独特的方式分布。这种分布表明,肌动蛋白除了作为一种结构蛋白发挥功能外,可能还参与上皮紧密连接通透性的调节、包括细胞从隐窝向绒毛顶端迁移的运动过程、对侵入性上皮内细胞的容纳以及细胞彼此之间及其与基质的黏附。

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