Kennedy J M, Kamel S, Tambone W W, Vrbova G, Zak R
J Cell Biol. 1986 Sep;103(3):977-83. doi: 10.1083/jcb.103.3.977.
Hypertrophy was produced in the anterior latissimus dorsi (ALD) muscle of 5-wk-old chickens by application of a load to the humerus. After 4 wk, hypertrophied ALD muscles were greater than 2.5 times heavier than contralateral control ALD muscles. Two isomyosins are distinguishable in normal ALD muscles by their different electrophoretic mobilities. It is shown here that the faster migrating SM-1 isomyosin decreases in abundance with age and that the application of an overload enhances both the rate and extent of this process. Monoclonal antibodies were selected by an immunotransfer technique that were specific for the heavy chains associated with either SM-1 or SM-2, or cross-reacted with both isoforms. The cellular distribution of the SM-1 and SM-2 isomyosins was analyzed by immunofluorescent technique using these antibodies. Anti-SM-1 and anti-SM-2 antibodies reacted with separate populations of cells, whereas the third antibody reacted with all myocytes in the normal ALD muscle. These data suggest that there is an exclusive cellular distribution of myosin heavy chains associated with SM-1 and SM-2 proteins. Immunofluorescent analysis of hypertrophied muscle showed the anti-SM-2-specific antibody reacting with all myocytes, whereas the anti-SM-1-specific antibody reacted with none. This is consistent with the elimination of the SM-1 isoform in hypertrophied muscles.
通过对5周龄鸡的肱骨施加负荷,在其背阔肌前束(ALD)中诱导产生肥大。4周后,肥大的ALD肌肉比同侧对照ALD肌肉重2.5倍以上。在正常的ALD肌肉中,两种同工肌球蛋白因其不同的电泳迁移率而可区分。本文表明,迁移速度较快的SM-1同工肌球蛋白的丰度随年龄增长而降低,施加过载会提高这一过程的速度和程度。通过免疫转移技术筛选出对与SM-1或SM-2相关的重链具有特异性、或与两种同工型均发生交叉反应的单克隆抗体。使用这些抗体,通过免疫荧光技术分析SM-1和SM-2同工肌球蛋白的细胞分布。抗SM-1和抗SM-2抗体与不同的细胞群体发生反应,而第三种抗体与正常ALD肌肉中的所有肌细胞发生反应。这些数据表明,与SM-1和SM-2蛋白相关的肌球蛋白重链在细胞中存在排他性分布。对肥大肌肉的免疫荧光分析显示,抗SM-2特异性抗体与所有肌细胞发生反应,而抗SM-1特异性抗体未与任何肌细胞发生反应。这与肥大肌肉中SM-1同工型的消除一致。