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鸡成肌细胞分化过程中醛缩酶A和醛缩酶C基因表达的稳态mRNA的非协同变化。

Noncoordinate changes in the steady-state mRNA expressed from aldolase A and aldolase C genes during differentiation of chicken myoblasts.

作者信息

Meighan-Mantha R L, Tolan D R

机构信息

Department of Biology, Boston University, Massachusetts 02215, USA.

出版信息

J Cell Biochem. 1995 Mar;57(3):423-31. doi: 10.1002/jcb.240570308.

Abstract

In chickens, as in all vertebrates, tissue-specific expression of aldolase isozymes A, B, and C is developmentally coordinated. These developmental transitions in aldolase expression have been studied most extensively by charting enzyme activity during normal and abnormal development of specific vertebrate tissues. Indeed, aldolase expression has been a key marker for normal differentiation and for retrodifferentiation during carcinogenesis. Aldolase expression during chicken myoblast differentiation offers a model for investigating the regulatory mechanisms of these developmental transitions at the level of gene expression. For these studies, cDNAs encoding the most isozyme-specific regions of both chicken aldolase A and C were cloned. The chicken aldolase A cDNA represents the first report of this sequence. Aldolase steady-state mRNA expression was measured during chicken myoblast differentiation in primary cultures using RNase protection assays with cRNA probes generated from these aldolase cDNA clones. Steady-state mRNA for aldolase C, the predominant embryonic aldolase isozyme in chickens, did not significantly change throughout myoblast differentiation. In contrast, expression of steady-state mRNA for aldolase A, the only aldolase isozyme found in adult-skeletal muscle, was not detected until after myoblast fusion was approximately 50% completed. Aldolase A expression gradually increased throughout myoblast differentiation until approximately 48 h after fusion was completed when there was a dramatic increase. These results are contrasted with those of Turner et al. (1974) [Dev Biol 37:63-89] that showed a coordinated switch in isozyme activities between the embryonic aldolase C and the muscle-specific aldolase A. This discordant expression indicates that the aldolase A and C genes may employ different regulatory mechanisms during myoblast differentiation.

摘要

与所有脊椎动物一样,鸡体内醛缩酶同工酶A、B和C的组织特异性表达在发育过程中是协调一致的。通过绘制特定脊椎动物组织在正常和异常发育过程中的酶活性,人们对醛缩酶表达的这些发育转变进行了最为广泛的研究。事实上,醛缩酶表达一直是正常分化以及癌变过程中逆分化的关键标志物。鸡成肌细胞分化过程中的醛缩酶表达为在基因表达水平上研究这些发育转变的调控机制提供了一个模型。为了进行这些研究,克隆了编码鸡醛缩酶A和C的最具同工酶特异性区域的cDNA。鸡醛缩酶A cDNA代表了该序列的首次报道。在原代培养的鸡成肌细胞分化过程中,使用从这些醛缩酶cDNA克隆产生的cRNA探针,通过核糖核酸酶保护试验测量醛缩酶的稳态mRNA表达。醛缩酶C是鸡体内主要的胚胎醛缩酶同工酶,其稳态mRNA在整个成肌细胞分化过程中没有显著变化。相比之下,醛缩酶A是成年骨骼肌中唯一发现的醛缩酶同工酶,其稳态mRNA直到成肌细胞融合完成约50%后才被检测到。醛缩酶A的表达在整个成肌细胞分化过程中逐渐增加,直到融合完成约48小时后出现急剧增加。这些结果与特纳等人(1974年)[《发育生物学》37:63 - 89]的结果形成对比,他们的研究表明胚胎醛缩酶C和肌肉特异性醛缩酶A之间的同工酶活性发生了协调转换。这种不一致的表达表明醛缩酶A和C基因在成肌细胞分化过程中可能采用不同的调控机制。

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