Sartore S, Gorza L, Pierobon Bormioli S, Dalla Libera L, Schiaffino S
J Cell Biol. 1981 Jan;88(1):226-33. doi: 10.1083/jcb.88.1.226.
Antisera against bovine atrial myosin were raised in rabbits, purified by affinity chromatography, and absorbed with insolubilized ventricular myosin. Specific anti-bovine atrial myosin (anti-bAm) antibodies reacted selectively with atrial myosin heavy chains, as determined by enzyme immunoassay combined with SDS-gel electrophoresis. In direct and indirect immunofluorescence assay, anti-bAm was found to stain all atrial muscle fibers and a minor proportion of ventricular muscle fibers in the right ventricle of the bovine heart. In contrast, almost all muscle fibers in the left ventricle were unreactive. Purkinje fibers showed variable reactivity. In the rabbit heart, all atrial muscle fibers were stained by anti-bAm, whereas ventricular fibers showed a variable response in both the right and left ventricle, with a tendency for reactive fibers to be more numerous in the right ventricle and in subepicardial regions. Diversification of fiber types with respect to anti-bAm reactivity was found to occur during late stages of postnatal development in the rabbit heart and to be influenced by thyroid hormone. All ventricular muscle fibers became strongly reactive after thyroxine treatment, whereas they became unreactive or poorly reactive after propylthiouracil treatment. These findings are consistent with the existence of different ventricular isomyosins whose relative proportions can vary according to the thyroid state. Variations in ventricular isomyosin composition can account for the changes in myosin Ca2+-activated ATPase activity previously observed in cardiac muscle from hyper- and hypothyroid animals and may be responsible for the changes in the velocity of contraction of ventricular myocardium that occur under these conditions. The differential distribution of ventricular isomyosins in the normal heart suggests that fiber types with different contractile properties may coexist in the ventricular myocardium.
用兔制备了抗牛心房肌球蛋白抗血清,通过亲和层析进行纯化,并用不溶性心室肌球蛋白进行吸附。通过酶免疫测定结合SDS凝胶电泳确定,特异性抗牛心房肌球蛋白(抗bAm)抗体与心房肌球蛋白重链发生选择性反应。在直接和间接免疫荧光测定中,发现抗bAm可使牛心脏右心室的所有心房肌纤维以及一小部分心室肌纤维着色。相比之下,左心室几乎所有的肌纤维都无反应。浦肯野纤维的反应性各不相同。在兔心脏中,所有心房肌纤维都被抗bAm染色,而心室纤维在右心室和左心室均表现出不同的反应,右心室和心外膜下区域的反应性纤维往往更多。发现兔心脏出生后发育后期,抗bAm反应性的纤维类型出现分化,且受甲状腺激素影响。甲状腺素治疗后,所有心室肌纤维都具有强反应性,而丙硫氧嘧啶治疗后,它们变得无反应或反应性很差。这些发现与不同心室同工肌球蛋白的存在一致,其相对比例可根据甲状腺状态而变化。心室同工肌球蛋白组成的变化可以解释先前在甲状腺功能亢进和减退动物的心肌中观察到的肌球蛋白Ca2+激活的ATP酶活性的变化,并且可能是这些条件下心室心肌收缩速度变化的原因。正常心脏中心室同工肌球蛋白的差异分布表明,具有不同收缩特性的纤维类型可能共存于心室心肌中。