COSMOS E
J Cell Biol. 1964 Nov;23(2):241-52. doi: 10.1083/jcb.23.2.241.
To follow the intracellular distribution of calcium in the breast muscles of developing chickens, Ca(45) was injected into the albumen of predeveloped eggs. Since the embryos were grown in a radioactive medium, a complete exchange of the isotope for its non-radioactive counterpart in muscles was accomplished. Subcellular particulates of the muscle cells were separated by the method of differential centrifugation. Analysis of the separated fractions showed that in the muscles of the 13-day embryo, when the nuclear-myofibrillar ratio is high, 65 per cent of the muscle calcium is in the nuclei. With the increased synthesis of myofibrils, the nuclear-myofibrillar ratio decreases with a concomitant fall in radioactivity. Thus, calcium was not associated with the developing myofibrils. At the time of hatching, when myofibrils perform physiological work, the highest level of calcium is in the mitochondria. This suggests that the mitochondria play a key role in the physiological activities of calcium in the cell. The microsomal fraction reaches a maximal level of calcium when the adult composition of muscle is attained. Results of investigations on dystrophic muscles show changes in the calcium distribution of the fractions as early as the 3rd week of embryonic development, which are interpreted to indicate an alteration in the protein metabolism of the cell, or an early destruction of muscle tissue. Further, alterations in the calcium content of fractions which seem to regulate the movements of this ion in the cell are discussed. A new technique for homogenizing tissues from embryos of different ages is presented.
为追踪发育中雏鸡胸肌内钙的细胞内分布情况,将Ca(45)注入发育前鸡蛋的蛋白中。由于胚胎在放射性培养基中生长,肌肉中的同位素与非放射性对应物实现了完全交换。通过差速离心法分离肌肉细胞的亚细胞颗粒。对分离出的组分进行分析表明,在13日龄胚胎的肌肉中,当核肌原纤维比例较高时,65%的肌肉钙存在于细胞核中。随着肌原纤维合成增加,核肌原纤维比例下降,放射性也随之降低。因此,钙与发育中的肌原纤维无关。在孵化时,当肌原纤维进行生理活动时,钙的最高水平存在于线粒体中。这表明线粒体在细胞内钙的生理活动中起关键作用。当肌肉达到成年组成时,微粒体组分中的钙达到最高水平。对营养不良肌肉的研究结果表明,早在胚胎发育的第3周,各组分的钙分布就发生了变化,这被解释为表明细胞蛋白质代谢发生了改变,或者肌肉组织早期遭到破坏。此外,还讨论了似乎调节细胞内该离子运动的组分中钙含量的变化。介绍了一种用于匀浆不同年龄胚胎组织的新技术。