Patapoutian A, Miner J H, Lyons G E, Wold B
Division of Biology, California Institute of Technology, Pasadena 91125.
Development. 1993 May;118(1):61-9. doi: 10.1242/dev.118.1.61.
In developing mouse embryos, MyoD family regulatory genes are expressed specifically in muscle precursors and mature myofibers. This pattern, taken together with the well-established ability of MyoD family members to convert a variety of cell types to skeletal muscle, suggests a significant role for these genes in regulating skeletal myogenesis. The possibility that expression of these genes may be causally associated with segregation of the myogenic lineage from other mesodermal derivatives, or with the subsequent maintenance of muscle phenotypes at later times, raises the issue of how MyoD family genes are themselves regulated during development. In this work, we have initiated studies to identify DNA sequences that govern Myf-5 and MRF4 (herculin, myf-6) transcription. Myf-5 is the first of the MyoD family to be expressed in the developing mouse embryo, while MRF4 is the most abundantly expressed myogenic factor in postnatal animals. In spite of their strikingly divergent patterns of expression, Myf-5 and MRF4 are tightly linked in the mouse genome; their translational start codons are only 8.5 kilobases apart. Here, the 5' flanking regions of the mouse Myf-5 and MRF4 genes were separately linked to a bacterial beta-galactosidase (lacZ) gene, and these constructs were each used to produce several lines of transgenic mice. Transgene expression was monitored by X-gal staining of whole embryos and by in situ hybridization of embryo sections. For the Myf-5/lacZ lines, the most intense transgene expression was in the visceral arches and their craniofacial muscle derivatives, beginning at day 8.75 post coitum (p.c.). This correlates with endogenous Myf-5 expression in visceral arches.(ABSTRACT TRUNCATED AT 250 WORDS)
在发育中的小鼠胚胎中,MyoD家族调控基因特异性地在肌肉前体细胞和成熟肌纤维中表达。这种表达模式,再加上MyoD家族成员将多种细胞类型转化为骨骼肌的既定能力,表明这些基因在调节骨骼肌生成中发挥着重要作用。这些基因的表达可能与肌源性谱系从其他中胚层衍生物中分离存在因果关系,或者与后期肌肉表型的维持有关,这就引发了MyoD家族基因在发育过程中自身如何被调控的问题。在这项研究中,我们开始了相关研究,以鉴定调控Myf-5和MRF4(赫库林,myf-6)转录的DNA序列。Myf-5是MyoD家族中第一个在发育中的小鼠胚胎中表达的基因,而MRF4是出生后动物中表达最丰富的肌源性因子。尽管Myf-5和MRF4的表达模式差异显著,但它们在小鼠基因组中紧密相连;它们的翻译起始密码子仅相隔8.5千碱基对。在这里,小鼠Myf-5和MRF4基因的5'侧翼区域分别与细菌β-半乳糖苷酶(lacZ)基因相连,并且这些构建体各自用于产生几系转基因小鼠。通过对整个胚胎进行X-gal染色以及对胚胎切片进行原位杂交来监测转基因表达。对于Myf-5/lacZ系,最强烈的转基因表达出现在内脏弓及其颅面肌肉衍生物中,从交配后第8.75天(p.c.)开始。这与内脏弓中内源性Myf-5的表达相关。(摘要截短于250字)