Krishan K, Morgan M J, Zhao W, Dhoot G K
Department of Basic Sciences, The Royal Veterinary College, London, UK.
J Muscle Res Cell Motil. 2000;21(6):527-36. doi: 10.1023/a:1026541803317.
We have cloned cDNA sequences of both rat and mouse slow troponin T gene. These sequences share a high level of homology with each other and with the human slow troponin T gene although we were unable to detect an alternatively spliced exon present at 3' end of human slow troponin T cDNA in either mouse or rat cDNAs. Northern blot analysis detected a high level expression of slow troponin T in adult mouse Soleus with a lower level expression in mixed postnatal skeletal muscles. Unlike late fetal and postnatal skeletal muscles in which slow troponin T expression is restricted to slow muscle fibre rich regions only, in situ hybridisation analysis detected this isoform to be highly expressed in somitic myotome and all muscle masses at 10-14 days of gestation after which its expression was rapidly downregulated. The unexpected expression of slow troponin T mRNA in fetal heart was apparent by both northern blotting and in situ hybridisation analyses. Slow troponin T mRNA in fetal heart was first detected at 10 day in utero reaching maximum levels of expression at 12-15 days gestation. The slow troponin T in the heart was mainly expressed in the ventral ventricles until day 15 after which low level expression was also observed in both atria. Slow troponin T mRNA in both atrium and ventricle was mainly expressed in outer wall of the myocardium although it was also expressed in interventricular septum. This study therefore shows that in addition to being a cell type specific marker during later fetal and postnatal skeletal muscle development, slow troponin T represented one of the major developmental isoforms expressed in embryonic and fetal skeletal muscle as well as in the cardiac muscle.
我们已经克隆了大鼠和小鼠慢肌钙蛋白T基因的cDNA序列。这些序列彼此之间以及与人类慢肌钙蛋白T基因具有高度同源性,尽管我们在小鼠或大鼠的cDNA中均未检测到人类慢肌钙蛋白T cDNA 3'端存在的可变剪接外显子。Northern印迹分析检测到慢肌钙蛋白T在成年小鼠比目鱼肌中高表达,而在出生后的混合骨骼肌中表达水平较低。与晚期胎儿和出生后的骨骼肌不同,在这些组织中慢肌钙蛋白T的表达仅限于富含慢肌纤维的区域,原位杂交分析检测到该异构体在妊娠10 - 14天时在体节肌节和所有肌肉团块中高表达,之后其表达迅速下调。通过Northern印迹和原位杂交分析均明显观察到胎儿心脏中慢肌钙蛋白T mRNA的意外表达。胎儿心脏中的慢肌钙蛋白T mRNA在子宫内10天时首次检测到,在妊娠12 - 15天时达到最高表达水平。心脏中的慢肌钙蛋白T主要在心室腹侧表达,直到第15天,之后在两个心房中也观察到低水平表达。心房和心室中的慢肌钙蛋白T mRNA主要在心肌外层表达,尽管在室间隔中也有表达。因此,这项研究表明,慢肌钙蛋白T除了是晚期胎儿和出生后骨骼肌发育过程中的细胞类型特异性标志物外,还是在胚胎和胎儿骨骼肌以及心肌中表达的主要发育异构体之一。