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伴刀豆球蛋白A受体和表面免疫球蛋白在大鼠肝细胞质膜上的流动性

The mobility of concanavalin A receptors and surface immunoglobulins on rat hepatocyte plasma membranes.

作者信息

Lustyik G, Kitani K, Ohta M

出版信息

Biochim Biophys Acta. 1987 Jan 9;896(1):57-63. doi: 10.1016/0005-2736(87)90356-7.

Abstract

Lateral mobilities of lectin receptors and surface immunoglobulins were measured in plasma membranes of hepatocytes prepared by smearing small pieces of rat liver tissue and then using the fluorescence recovery after photobleaching (FRAP) technique. Smears were treated with various doses of fluorescein isothiocyanate (FITC) conjugated concanavalin A (ConA), succinylated ConA (SConA), wheat germ agglutinin (WGA), and soybean agglutinin (SBA), as well as with rabbit anti-rat IgG (RARa/IgG) and goat anti-rat IgM(Fc) (GARa/IgM(Fc] antisera. 10 micrograms/ml ConA and SConA concentrations and a 55 X dilution of the GARa/IgM(Fc) antiserum were found to be suitable for measuring the lateral mobilities dependent on age. Diffusion constant and mobile fractions of receptor complexes were measured in different age groups of female Fisher rats (from 1 to 26 month-old). The FRAP measurements revealed that at least two major receptor sites can be distinguished in cell membranes of compact tissue (similar to the cultured and isolated cells), forming a mobile and an immobile fraction. The mobile fractions of both the lectin receptors and the surface immunoglobulins tended to decrease with age, while the age differences of the diffusion constants were not statistically significant. The observed alterations could be due to the covalent crosslinking of the mobile receptors to immobile patches and/or to the retardation of free diffusion by the cytoskeleton, dependent on age.

摘要

通过涂抹大鼠肝脏组织小块,然后使用光漂白后荧光恢复(FRAP)技术制备肝细胞质膜,测量凝集素受体和表面免疫球蛋白的侧向迁移率。涂片用不同剂量的异硫氰酸荧光素(FITC)偶联的伴刀豆球蛋白A(ConA)、琥珀酰化ConA(SConA)、麦胚凝集素(WGA)和大豆凝集素(SBA)处理,以及用兔抗大鼠IgG(RARa/IgG)和山羊抗大鼠IgM(Fc)(GARa/IgM(Fc)抗血清处理。发现10微克/毫升的ConA和SConA浓度以及GARa/IgM(Fc)抗血清的55倍稀释度适合测量与年龄相关的侧向迁移率。在不同年龄组的雌性Fisher大鼠(1至26月龄)中测量受体复合物的扩散常数和可移动部分。FRAP测量显示,在致密组织的细胞膜中(类似于培养和分离的细胞)至少可以区分两个主要的受体位点,形成一个可移动部分和一个不可移动部分。凝集素受体和表面免疫球蛋白的可移动部分均随年龄增长而趋于下降,而扩散常数的年龄差异无统计学意义。观察到的变化可能是由于可移动受体与不可移动斑块的共价交联和/或细胞骨架对自由扩散的阻碍,这取决于年龄。

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