Kohnen G, Castellucci M, Hsi B L, Yeh C J, Kaufmann P
Department of Anatomy, Technical University of Aachen, Germany.
Cell Tissue Res. 1995 Aug;281(2):231-42. doi: 10.1007/BF00583392.
The expression patterns of a variety of cytoskeletal antigens were studied in normal human tissues (placenta, umbilical cord, myometrium, colon, mammary gland, testis, skeletal muscle, myocardium) as well as in abnormal human tissues (palmar fibromatosis, fibrocystic disease of the mammary gland, mammary carcinoma). The immunohistochemical binding patterns of the monoclonal antibody GB 42 were compared to those of commercial antibodies directed against vimentin, desmin, smooth muscle myosin, pan actin, alpha-smooth muscle actin and gamma-smooth muscle actin. Methods applied comprised immunohistochemistry on cryostat sections and paraffin sections. Immunogold immunocytochemistry was performed on Lowicryl sections. The patterns of GB 42-binding were confirmed biochemically by SDS-PAGE and Western-blotting, and quantitative amino acid analysis. Our data suggest that the monoclonal antibody GB 42 recognizes an actin isoform which is identical to, or closely related to, gamma-smooth muscle actin. Unlike the commercially available antibody against gamma-smooth muscle actin, GB 42 does not cross-react with alpha-skeletal or alpha-cardiac actins. The GB 42-antigen is expressed in smooth muscle cells, myoepithelial cells and in later stages of differentiation of myofibroblasts, in all the tissues investigated. Throughout the development of smooth muscle cells and myofibroblasts, the appearance of the GB 42-antigen occurs after the expression of vimentin, desmin and alpha-smooth muscle actin, but prior to the expression of smooth muscle myosin. GB 42 is a reliable marker for higher stages of differentiation of smooth muscle cells and myofibroblasts.
研究了多种细胞骨架抗原在正常人体组织（胎盘、脐带、子宫肌层、结肠、乳腺、睾丸、骨骼肌、心肌）以及异常人体组织（掌纤维瘤病、乳腺纤维囊性疾病、乳腺癌）中的表达模式。将单克隆抗体GB 42的免疫组化结合模式与针对波形蛋白、结蛋白、平滑肌肌球蛋白、泛肌动蛋白、α-平滑肌肌动蛋白和γ-平滑肌肌动蛋白的商用抗体的结合模式进行了比较。应用的方法包括对冷冻切片和石蜡切片进行免疫组化。对Lowicryl切片进行免疫金免疫细胞化学。通过SDS-PAGE、蛋白质印迹和定量氨基酸分析对GB 42结合模式进行了生化确认。我们的数据表明，单克隆抗体GB 42识别一种肌动蛋白异构体，它与γ-平滑肌肌动蛋白相同或密切相关。与市售的抗γ-平滑肌肌动蛋白抗体不同，GB 42不与α-骨骼肌肌动蛋白或α-心肌肌动蛋白发生交叉反应。GB 42抗原在所有研究的组织中的平滑肌细胞、肌上皮细胞和成肌纤维细胞分化的后期阶段表达。在平滑肌细胞和成肌纤维细胞的整个发育过程中，GB 42抗原的出现发生在波形蛋白、结蛋白和α-平滑肌肌动蛋白表达之后，但在平滑肌肌球蛋白表达之前。GB 42是平滑肌细胞和成肌纤维细胞较高分化阶段的可靠标志物。
