Kaufman S J, Foster R F, Haye K R, Faiman L E
J Cell Biol. 1985 Jun;100(6):1977-87. doi: 10.1083/jcb.100.6.1977.
H36 is a species-specific, cell-surface antigen on differentiating newborn rat skeletal myoblasts and myogenic lines. This membrane antigen has been defined by a monoclonal antibody raised by the fusion of SP 2/0-Ag14 myeloma cells with spleen cells from mice immunized with myotubes derived from the myogenic E63 line. H36 antigen, isolated by immunoaffinity chromatography, is comprised of two polypeptides with apparent molecular weights of 98,000 and 117,000. Fluorescence photometry and radioimmunoassays have been used to follow quantitative and topographic changes in the H36 determinant during myogenesis. H36 is present at a basal level on replicating myoblasts; it increases on prefusion myoblasts and persists on myotubes. At or near the time of prefusion, it becomes concentrated between adjacent aligned myoblasts and localized on membrane "blebs". H36 is present on both skeletal and cardiac cells but absent from a variety of cells that include fibroblasts, neuronal cells, and smooth muscle. There are approximately 4 x 10(5) determinants per myoblast, and the Ka of the antibody is 3.8 x 10(8) liters/mol. The distributions of H36 on the top and attached surfaces of myoblasts and myotubes are distinct, which suggests localized specialization of these surfaces. H36 is an integral membrane component and upon cross-linking, it associates with the detergent-insoluble cytoskeletal framework. Inhibition of myogenesis by 5-bromodeoxyuridine or by calcium deprivation prevents the developmentally associated changes in the expression of H36. H36 is also absent or markedly reduced on the fu- and Ama102 developmentally defective mutant myoblast lines. We conclude that H36 is a muscle-specific, developmentally regulated cell-surface antigen that may have a role in myoblast differentiation and that can be used to determine the embryonic lineages of skeletal and cardiac muscle.
H36是新生大鼠分化中的骨骼肌成肌细胞及成肌细胞系上一种物种特异性的细胞表面抗原。这种膜抗原是由SP 2/0-Ag14骨髓瘤细胞与用源自成肌E63细胞系的肌管免疫的小鼠脾细胞融合产生的单克隆抗体所界定。通过免疫亲和层析分离得到的H36抗原由两条表观分子量分别为98,000和117,000的多肽组成。荧光光度法和放射免疫测定法已被用于追踪成肌过程中H36决定簇的定量及拓扑学变化。H36在增殖的成肌细胞上以基础水平存在;在融合前的成肌细胞上增加,并在肌管上持续存在。在融合前或融合时,它在相邻排列的成肌细胞之间聚集,并定位在膜“泡”上。H36存在于骨骼肌细胞和心肌细胞上,但在包括成纤维细胞、神经元细胞和平滑肌在内的多种细胞中不存在。每个成肌细胞约有4×10(5)个决定簇,抗体的Ka为3.8×10(8)升/摩尔。H36在成肌细胞和肌管的顶部及附着表面的分布不同,这表明这些表面存在局部特化。H36是一种整合膜成分,交联后与去污剂不溶性细胞骨架框架相关联。5-溴脱氧尿苷或钙缺乏对成肌的抑制作用可防止H36表达中与发育相关的变化。在发育缺陷的突变成肌细胞系fu和Ama102上,H36也不存在或显著减少。我们得出结论,H36是一种肌肉特异性、发育调控的细胞表面抗原,可能在成肌细胞分化中起作用,并且可用于确定骨骼肌和心肌的胚胎谱系。