Presence of laminin alpha 5 chain and lack of laminin alpha 1 chain during human muscle development and in muscular dystrophies
Presence of laminin alpha 5 chain and lack of laminin alpha 1 chain during human muscle development and in muscular dystrophies
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DOI：
10.1074/jbc.272.45.28590
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发表时间：
1997-11-07
影响因子：
4.8
通讯作者：
Gullberg, D
中科院分区：
文献类型：
--
作者：
Tiger, CF;Champliaud, MF;Gullberg, D
目前，人们对鉴定在发育和再生骨骼肌中表达的层粘连蛋白亚型非常感兴趣。层粘连蛋白α1已被报道定位于人胎儿肌肉，并在肌营养不良症中被诱导，建议识别人层粘连蛋白α1链。然而，在发育中或营养不良的小鼠骨骼肌纤维中似乎没有层粘连蛋白α1蛋白或mRNA的表达。为了解决发育中的人和营养不良的人和小鼠肌肉的结果之间的差异，我们以重组蛋白的形式表达了人层粘连蛋白α1链的E3结构域，并制备了针对人层粘连蛋白α1链的抗体(抗HLN-α1G4/G5)。我们还从胎盘中提纯了人层粘连蛋白α5链，并制备了抗体。在本报告中，我们发现HLN-α1G4/G5抗体与400 kDa的层粘连蛋白α1链反应，4C7与380 kDa的层粘连蛋白α5链反应。HLN-α1G4/G5抗体和4C7免疫组织化学染色显示，这两种抗体对人肾脏、发育中和营养不良的肌肉有不同的染色模式。我们的数据表明，先前报道的4C7在发育中、成人和营养不良的人类肌肉组织中的表达模式应该被重新解释为层粘连蛋白α5链的表达。我们的数据也与小鼠早期的工作一致，表明层粘连蛋白α1主要是一种上皮层粘连蛋白链，在发育中或营养不良的肌肉纤维中不存在。
There is currently a great interest in identifying laminin isoforms expressed in developing and regenerating skeletal muscle. Laminin alpha 1 has been reported to localize to human fetal muscle and to be induced in muscular dystrophies based on immunohistochemistry with the monoclonal antibody 4C7, suggested to recognize the human laminin alpha 1 chain. Nevertheless, there seems to be no expression of laminin alpha 1 protein or mRNA in developing or dystrophic mouse skeletal muscle fibers. To address the discrepancy between the results obtained in developing and dystrophic human and mouse muscle we expressed the E3 domain of human laminin alpha 1 chain as a recombinant protein and made antibodies specific for human laminin alpha 1 chain (anti-hLN-alpha 1G4/G5). We also made antibodies to the human laminin alpha 5 chain purified from placenta. In the present report we show that hLN-alpha 1G4/G5 antibodies react with a 400-kDa laminin alpha 1 chain and that 4C7 reacts with a 380-kDa laminin alpha 5 chain. Immunohistochemistry with the hLN-alpha 1G4/G5 antibody and 4C7 revealed that the two antibodies stained human kidney, developing and dystrophic muscle in distinct patterns. Our data indicate that the previously reported expression patterns in developing, adult, and dystrophic human muscle tissues with 4C7 should be re-interpreted as an expression of laminin alpha 5 chain. Our data are also consistent with earlier work in mouse, indicating that laminin alpha 1 is largely an epithelial laminin chain not present in developing or dystrophic muscle fibers.