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钙调蛋白在仓鼠气管上皮纤毛细胞中的免疫定位。

Immune localization of calmodulin in the ciliated cells of hamster tracheal epithelium.

作者信息

Gordon R E, Williams K B, Puszkin S

出版信息

J Cell Biol. 1982 Oct;95(1):57-63. doi: 10.1083/jcb.95.1.57.

Abstract

Melachronous beating of cilia of epithelial surfaces of most respiratory airways moves the overlying mucous layer in a caudal direction. The molecular mechanisms controlling ciliary beat remain largely unknown. Calcium, an element in its cationic form, is ubiquitous in biological functions and its concentration is critical for ciliary beating. Calmodulin, a calcium-binding protein which regulates the activity of many enzymes and cellular processes, may regulate ciliary beating by controlling enzymes responsible for mechanochemical movement between adjacent peripheral microtubule doublets composing the ciliary axoneme. As a first step in describing a calmodulin-related controlling mechanism for ciliary beating, calmodulin was localized in the ciliated cells lining the respiratory tracts of hamsters by electron microscopy, using an indirect immunoperoxidase technique with anticalmodulin antibodies as the molecular probe. Thin-sections revealed calmodulin located on microtubules and dynein arms of the ciliary shaft, basal body, apical cytoskeletal microtubules, and plasma membranes in specimens fixed with 1 mM Ca+2. Specimens fixed with less Ca+2 (1 microM), Mn+2, Mg+2, and EGTA showed a diffuse pattern of calmodulin with loci of greatest densities on basal body microtubule triplets. Demembranated specimens showed a less specific localization on axonemal microtubules but only on cells fixed with Ca+2. Calmodulin, by binding calcium, may function in ciliary beating in the respiratory tract of mammals either directly or indirectly through its effects on the energy-producing enzymes and by control of Ca+2 flux through plasma membranes.

摘要

大多数呼吸道上皮表面的纤毛进行异步摆动,将覆盖其上的黏液层向尾端方向移动。控制纤毛摆动的分子机制在很大程度上仍不为人知。钙,以阳离子形式存在的一种元素,在生物功能中普遍存在,其浓度对纤毛摆动至关重要。钙调蛋白是一种钙结合蛋白,可调节许多酶的活性和细胞过程,它可能通过控制负责组成纤毛轴丝的相邻外周微管双联体之间机械化学运动的酶来调节纤毛摆动。作为描述纤毛摆动的钙调蛋白相关控制机制的第一步,利用以抗钙调蛋白抗体作为分子探针的间接免疫过氧化物酶技术,通过电子显微镜将钙调蛋白定位在仓鼠呼吸道的纤毛细胞中。超薄切片显示,在用1 mM Ca+2固定的标本中,钙调蛋白位于纤毛轴、基体、顶端细胞骨架微管和质膜的微管和动力蛋白臂上。用较少Ca+2(1 microM)、Mn+2、Mg+2和EGTA固定的标本显示钙调蛋白呈弥散分布模式,在基体微管三联体上密度最大。去膜标本在轴丝微管上的定位不太特异,但仅在Ca+2固定的细胞上有定位。钙调蛋白通过结合钙,可能在哺乳动物呼吸道的纤毛摆动中直接或间接发挥作用,其作用途径是通过影响产生能量的酶以及控制Ca+2通过质膜的通量。

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