González-Sánchez A, Bader D
J Cell Biol. 1985 Jan;100(1):270-5. doi: 10.1083/jcb.100.1.270.
A monoclonal antibody (anterior latissimus dorsi 58 [ALD58]; antimyosin heavy chain, MHC) directed against myosin from slow tonic muscle was found to react specifically with the striated muscle cells of the conductive system in the adult chicken heart. This monoclonal antibody was used to study the expression of myosin in the conductive system of the adult and developing heart. Using immunofluorescence microscopy with ALD58, muscle cells of the conductive system were demonstrated in both the atria and ventricles of the adult heart as previously shown by Sartore et al. (Sartore, S., S. Pierobon-Bormioli, and S. Schiafinno, 1978, Nature (Lond.), 274: 82-83). Radioactive myosin from adult atria and ventricles was precipitated with ALD58 and subjected to limited proteolysis and subsequent peptide mapping. Peptide maps of ALD58 reactive myosin from atria and ventricles were very similar, if not identical, but differed from peptide maps of ordinary atrial and ventricular myosin. The same antibody was used to study cardiac myogenesis in the chick embryo. When ALD58 was reacted with myosin isolated from atria and ventricles at selected stages of development in radioimmunoassays, reactivity was not observed until the last week of embryonic life (greater than 15 d of egg incubation). Thereafter concomitant and progressively increased reactivity was observed in atrial and ventricular preparations. Also, no ALD58 positive cells were observed in immunofluorescence studies of embryonic hearts until 17 d of egg incubation. Primary cell cultures of embryonic hearts also proved to be negative for this antibody. This study demonstrates that an epitope recognized by ALD58 associated with an antimyosin heavy chain of striated muscle cells of the adult heart conductive system is absent or present in only small amounts in the early embryonic heart.
一种针对慢张力肌肌球蛋白的单克隆抗体(背阔肌前部58 [ALD58];抗肌球蛋白重链,MHC)被发现能与成年鸡心脏传导系统的横纹肌细胞发生特异性反应。该单克隆抗体被用于研究成年和发育中心脏传导系统中肌球蛋白的表达。使用ALD58进行免疫荧光显微镜观察,如Sartore等人之前所示(Sartore, S., S. Pierobon - Bormioli, and S. Schiafinno, 1978, Nature (Lond.), 274: 82 - 83),在成年心脏的心房和心室中均证实了传导系统的肌细胞。来自成年心房和心室的放射性肌球蛋白用ALD58沉淀,进行有限的蛋白水解及随后的肽图谱分析。来自心房和心室的ALD58反应性肌球蛋白的肽图谱即使不完全相同也非常相似,但与普通心房和心室肌球蛋白的肽图谱不同。同一抗体被用于研究鸡胚心肌发生。当在放射免疫分析中使ALD58与发育特定阶段从心房和心室分离的肌球蛋白反应时,直到胚胎期最后一周(孵化超过15天)才观察到反应性。此后,在心房和心室标本中观察到伴随且逐渐增加的反应性。同样,在胚胎心脏的免疫荧光研究中,直到孵化17天时才观察到ALD58阳性细胞。胚胎心脏的原代细胞培养物对该抗体也呈阴性。这项研究表明,与成年心脏传导系统横纹肌细胞抗肌球蛋白重链相关的ALD58识别的表位在胚胎早期心脏中不存在或仅少量存在。