Samuel J L, Schwartz K, Lompre A M, Delcayre C, Marotte F, Swynghedauw B, Rappaport L
Eur J Cell Biol. 1983 Jul;31(1):99-106.
Isolated myocytes were purified from adult rat heart. Identification and localization of microtubules and quantitation of tubulin in these cells were performed by immunochemical procedures. Antibodies were raised against brain tubulin and purified by affinity chromatography. An enzyme-linked immunosorbent assay, ELISA, was developed for quantitation of tubulin. It allowed the measurement of 10 to 500 ng of tubulin. Tubulin content in adult rat cardiac myocytes was found to be approximately 10 micrograms per 100 mg of the total protein content. By means of a double immunofluorescence technique, the microtubule network, identified with antitubulin, was studied in reference to the sarcomeric A band labeled with antibodies specific to myosin heavy chains. The basis for identifying the microtubule network have included the use of specific antitubulin immunoglobulins and the sensitivity of the specific labeling of the network to antimitotic drugs and low temperature. It was found that microtubules were organized mainly around the nuclei, with important concentrations at the poles, showing extensions in the cone and in the cytoplasm as loosely organized loops. The shape of adult cardiac myocyte was not dependent upon the integrity of the microtubule network.
从成年大鼠心脏中分离出单个心肌细胞。通过免疫化学方法对这些细胞中的微管进行鉴定和定位，并对微管蛋白进行定量分析。制备了针对脑微管蛋白的抗体，并通过亲和层析进行纯化。开发了一种酶联免疫吸附测定法（ELISA）用于微管蛋白的定量分析。该方法可测量10至500纳克的微管蛋白。发现成年大鼠心肌细胞中的微管蛋白含量约为每100毫克总蛋白含量10微克。借助双重免疫荧光技术，以抗微管蛋白鉴定微管网络，并参照用肌球蛋白重链特异性抗体标记的肌节A带进行研究。鉴定微管网络的依据包括使用特异性抗微管蛋白免疫球蛋白，以及该网络的特异性标记对抗有丝分裂药物和低温的敏感性。结果发现，微管主要围绕细胞核排列，在两极有重要的聚集，在圆锥体和细胞质中呈松散组织的环延伸。成年心肌细胞的形状并不依赖于微管网络的完整性。
