Thézé N, Hardy S, Wilson R, Allo M R, Mohun T, Thiebaud P
Département de Biologie et Génétique du Développement, URA CNRS 256, Université de Rennes I, France.
Dev Biol. 1995 Oct;171(2):352-62. doi: 10.1006/dbio.1995.1287.
cDNAs clones encoding the MLC1f and MLC3f proteins of Xenopus laevis have been isolated from a stage 42 cDNA library. Sequence analysis reveals that the amphibian MLC1f and MLC3f isoforms are similar to the mammalian and avian cognates. The two isoforms share a common 141-amino-acid carboxy-terminal regions. These are 49 and 9 residues long for the MLC1f and MLC3f isoforms, respectively. This suggests a genomic organization similar to the mammalian and avian genes, with two promoters and alternative splicing. The developmental expression of the MLC1f/3f mRNAs was studied by Northern blot and RNase protection and their spatial expression analyzed by in situ hybridization. Both the MLC1f and MLC3f mRNAs can be detected in the developing embryo from the end of gastrulation and accumulate rapidly in the somitic mesoderm. Expression of the MLC1f/3f gene can also be detected in animal cap explants which have been induced to form mesodermal derivatives by exposure to activin A or bFGF. However, unlike other muscle-specific markers, neither transcript from the MLC1f/3f gene can be detected in embryonic or adult cardiac muscle, their expression being restricted to somitic muscle. Together, these data demonstrate that expression of the MLC1f/3f gene provides a sensitive and specific marker for skeletal muscle differentiation. Ectopic expression of myogenic factors in animal caps induces the expression of the MLC1f/3f gene, suggesting that the amphibian gene, like its mammalian and avian counterparts, is a regulatory target for members of the MyoD family of transcription factors.
已从非洲爪蟾第42期cDNA文库中分离出编码非洲爪蟾MLC1f和MLC3f蛋白的cDNA克隆。序列分析表明,两栖动物的MLC1f和MLC3f亚型与哺乳动物和鸟类的同源物相似。这两种亚型共享一个141个氨基酸的共同羧基末端区域。对于MLC1f和MLC3f亚型,这些区域分别长49个和9个残基。这表明其基因组组织类似于哺乳动物和鸟类基因,具有两个启动子和可变剪接。通过Northern印迹和RNA酶保护研究了MLC1f/3f mRNA的发育表达,并通过原位杂交分析了它们的空间表达。在原肠胚形成末期的发育胚胎中可检测到MLC1f和MLC3f mRNA,并且它们在体节中胚层中迅速积累。在通过暴露于激活素A或碱性成纤维细胞生长因子而被诱导形成中胚层衍生物的动物帽外植体中也可检测到MLC1f/3f基因的表达。然而,与其他肌肉特异性标志物不同,在胚胎或成年心肌中均未检测到MLC