Berezowsky C, Bag J
University of Guelph, Department of Molecular Biology and Genetics, Ont., Canada.
Biochem Cell Biol. 1988 Aug;66(8):880-8. doi: 10.1139/o88-100.
Regulation of slow troponin C gene expression was examined in both skeletal and cardiac muscle at various stages of development in chicken. The steady-state levels of troponin C mRNA were initially measured by Northern blot analysis. It was observed that the level of troponin C mRNA reached its maximum in both skeletal and cardiac muscle of 16- to 18-day-old embryos. A drop in troponin C mRNA level was observed just prior to hatching. The level of actin mRNA, myosin heavy chain mRNA, and mRNA for a nonmuscle protein, vimentin, was also similarly regulated during development of chicken muscles. Further studies were carried out to determine the level of slow troponin C mRNA using nuclease S1 protection analysis. A significant amount of slow troponin C mRNA was found in the skeletal muscle of adult chicken, which predominantly consists of the fast isoform of troponin C. This observation suggests the possibility of post-transcriptional control of slow troponin C synthesis in skeletal muscle. Primary cultures of cardiac myocytes were also used to determine how the troponin C mRNA level is regulated in a culture of cardiac muscle cells. Measurements of the steady-state levels of slow troponin C mRNA by nuclease S1 protection analysis show that it was maximal in 60-h-old cultures. A drop in the level of this mRNA was observed after these cells were maintained in culture for 4 days.
在鸡发育的各个阶段,对骨骼肌和心肌中慢肌钙蛋白C基因表达的调控进行了研究。最初通过Northern印迹分析测量肌钙蛋白C mRNA的稳态水平。观察到,在16至18日龄胚胎的骨骼肌和心肌中,肌钙蛋白C mRNA的水平均达到最高。在孵化前观察到肌钙蛋白C mRNA水平下降。在鸡肌肉发育过程中,肌动蛋白mRNA、肌球蛋白重链mRNA以及一种非肌肉蛋白波形蛋白的mRNA水平也受到类似的调控。使用核酸酶S1保护分析进行了进一步研究,以确定慢肌钙蛋白C mRNA的水平。在成年鸡的骨骼肌中发现了大量的慢肌钙蛋白C mRNA,而成年鸡骨骼肌主要由肌钙蛋白C的快异构体组成。这一观察结果表明,骨骼肌中慢肌钙蛋白C合成可能存在转录后调控。心肌细胞的原代培养也用于确定在心肌细胞培养物中肌钙蛋白C mRNA水平是如何调控的。通过核酸酶S1保护分析测量慢肌钙蛋白C mRNA的稳态水平,结果表明在60小时龄的培养物中其水平最高。在这些细胞在培养物中维持4天后,观察到这种mRNA水平下降。