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用HVJ(仙台病毒)M蛋白抗原性增强的抗大鼠脑肌动蛋白单克隆抗体。

Monoclonal antibody to rat brain actin antigenically enhanced with HVJ (Sendai virus) M protein.

作者信息

Hiruma S, Hashimoto S

机构信息

Second Department of Pathology, Kinki University School of Medicine, Osaka, Japan.

出版信息

Histochemistry. 1988;90(1):1-8. doi: 10.1007/BF00495699.

Abstract

Monoclonal antibody to rat brain actin was easily produced using HVJ (Sendai Virus) M protein to enhance the antigenicity of the actin. This monoclonal antibody was determined to be IgM with a kappa light chain. By immunoblot analysis the antibody was also shown to react with rat brain actin but not with HVJ M protein on nitrocellulose sheets. Utilizing the antibody, neuronal cytoplasm in the cerebral cortex, the anterior and posterior horns in the spinal cord, the spinal ganglion and astrocytes showed positive immunohistochemical staining by light microscopy. However, Purkinje cells showed variable staining, some staining intensely, while others were negative. All of neurons in specific anatomical locations showed always positive staining but variable intensities. Vascular walls were stained only faintly. By electron microscopy, neuronal cytoplasm showed diffuse positive staining. Other areas showed a positive reaction, including dendrites, the postsynaptic densities, and a few capillary endothelial cells and arterial smooth muscle cells. The results suggest that the HVJ M protein was effective for producing monoclonal antibody to brain actin, and that the antibody could be utilized for the immunohistochemical study of neuronal elements in both normal and pathological conditions.

摘要

利用仙台病毒(HVJ)M蛋白增强肌动蛋白的抗原性,很容易制备出针对大鼠脑肌动蛋白的单克隆抗体。该单克隆抗体被确定为具有κ轻链的IgM。通过免疫印迹分析,该抗体在硝酸纤维素膜上也显示与大鼠脑肌动蛋白发生反应,但不与HVJ M蛋白反应。利用该抗体,大脑皮质的神经元细胞质、脊髓的前后角、脊髓神经节和星形胶质细胞在光学显微镜下显示出阳性免疫组化染色。然而,浦肯野细胞染色情况不一,有些染色强烈,而有些则为阴性。特定解剖位置的所有神经元均显示始终呈阳性染色,但强度不同。血管壁仅呈微弱染色。通过电子显微镜观察,神经元细胞质显示弥漫性阳性染色。其他区域也呈阳性反应,包括树突、突触后致密物以及一些毛细血管内皮细胞和动脉平滑肌细胞。结果表明,HVJ M蛋白对于制备针对脑肌动蛋白的单克隆抗体是有效的,并且该抗体可用于正常和病理条件下神经元成分的免疫组化研究。

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