Gorza L, Ausoni S, Merciai N, Hastings K E, Schiaffino S
Department of Biomedical Sciences, University of Padova, Italy.
Dev Biol. 1993 Mar;156(1):253-64. doi: 10.1006/dbio.1993.1074.
Expression of cardiac troponin I (TnIcardiac) and slow skeletal troponin I (TnIslow) genes was analyzed at the mRNA and protein level in the developing rat heart. TnIslow mRNA was detectable by in situ hybridization in the embryonic cardiac tube as early as the 13-somite stage (Embryonic Day 10). In contrast, TnIcardiac transcripts were first detected in the primordial atrium and ventricle of 11-day-old embryos, but were absent in the outflow tract region. TnIslow mRNA levels decreased after birth in atria and later in ventricles but persisted even in adult life in myocytes of the conduction system. TnIslow protein was detected by specific antibodies in atrial myocytes beginning from Embryonic Day 11; in contrast, ventricular myocytes were unreactive until Embryonic Day 18. Western blot analysis of 16-day-old fetal hearts confirmed the expression of TnIcardiac in atrial but not in ventricular myocardium. Slot blot analysis showed that at this stage equivalent amounts of TnIslow and TnIcardiac mRNAs are expressed in atria and ventricles. Similar differences in the expression of TnIslow and TnIcardiac mRNAs and proteins were observed in cultures of embryonic atrial and ventricular myocytes. The results suggest serial rather than simultaneous activation of TnIslow and TnIcardiac genes and they show that different regions of the developing heart differ in their patterns of TnIcardiac expression due to the operation of distinct mechanisms that separately affect the accumulation of TnIcardiac mRNA and protein.
在发育中的大鼠心脏中,从mRNA和蛋白质水平分析了心肌肌钙蛋白I(TnIcardiac)和慢骨骼肌肌钙蛋白I(TnIslow)基因的表达。早在13体节期(胚胎第10天),通过原位杂交在胚胎心管中就可检测到TnIslow mRNA。相比之下,TnIcardiac转录本最早在11日龄胚胎的原始心房和心室中检测到,但在流出道区域不存在。出生后,心房中TnIslow mRNA水平下降,随后心室中也下降,但在传导系统的心肌细胞中即使在成年期仍持续存在。从胚胎第11天开始,在心房肌细胞中通过特异性抗体检测到TnIslow蛋白;相比之下,心室肌细胞直到胚胎第18天才有反应。对16日龄胎儿心脏的蛋白质印迹分析证实了TnIcardiac在心房心肌而非心室心肌中的表达。狭缝印迹分析表明,在此阶段,心房和心室中表达的TnIslow和TnIcardiac mRNA量相当。在胚胎心房和心室肌细胞培养物中也观察到TnIslow和TnIcardiac mRNA及蛋白质表达的类似差异。结果表明TnIslow和TnIcardiac基因是相继而非同时被激活的,并且它们表明,由于分别影响TnIcardiac mRNA和蛋白质积累的不同机制的作用,发育中心脏的不同区域在TnIcardiac表达模式上存在差异。