Macqueen Daniel J, Robb David, Johnston Ian A
Gatty Marine Laboratory, School of Biology, University of St Andrews, St Andrews, Fife, KY16 8LB, UK.
J Exp Biol. 2007 Aug;210(Pt 16):2781-94. doi: 10.1242/jeb.006981.
Potential molecular mechanisms regulating developmental plasticity to temperature were investigated in Atlantic salmon embryos (Salmo salar L.). Six orthologues of the four myogenic regulatory factors (MRFs: individually: smyf5, smyoD1a/1b/1c, smyoG and sMRF4), the master transcription factors regulating vertebrate myogenesis, were characterised at the mRNA/genomic level. In situ hybridisation was performed with specific cRNA probes to determine the expression patterns of each gene during embryonic myogenesis. To place the MRF data in the context of known muscle fibre differentiation events, the expression of slow myosin light chain-1 and Pax7 were also investigated. Adaxial myoblasts expressed smyoD1a prior to and during somitogenesis followed by smyoD1c (20-somite stage, ss), and sMRF4 (25-30 ss), before spreading laterally across the myotome, followed closely by the adaxial cells. Smyf5 was detected prior to somitogenesis, but not in the adaxial cells in contrast to other teleosts studied. The expression domains of smyf5, smyoD1b and smyoG were not confined to the s-smlc1 expression field, indicating a role in fast muscle myogenesis. From the end of segmentation, each MRF was expressed to a greater or lesser extent in zones of new muscle fibre production, the precursor cells for which probably originated from the Pax7 expressing cell layer external to the single layer of s-smlc1(+) fibres. SmyoD1a and smyoG showed similar expression patterns with respect to somite stage at three different temperatures investigated (2 degrees C, 5 degrees C and 8 degrees C) in spite of different rates of somite formation (one somite added each 5 h, 8 h and 15 h at 8 degrees C, 5 degrees C and 2 degrees C, respectively). In contrast, the expression of smyf5, sMRF4 and s-smlc1 was retarded with respect to somite stage at 2 degrees C compared to 8 degrees C, potentially resulting in heterochronies in downstream pathways influencing later muscle phenotype.
在大西洋鲑鱼胚胎(Salmo salar L.)中研究了调节发育可塑性以适应温度的潜在分子机制。对调节脊椎动物肌肉生成的主要转录因子——四个生肌调节因子(MRFs:分别为smyf5、smyoD1a/1b/1c、smyoG和sMRF4)的六个直系同源物进行了mRNA/基因组水平的表征。使用特异性cRNA探针进行原位杂交,以确定每个基因在胚胎肌肉生成过程中的表达模式。为了将MRF数据置于已知的肌纤维分化事件背景下,还研究了慢肌球蛋白轻链-1和Pax7的表达。在体节发生之前和期间,轴旁成肌细胞表达smyoD1a,随后在20体节期(ss)表达smyoD1c,在25 - 30体节期(ss)表达sMRF4,然后横向扩散到整个肌节,轴旁细胞紧随其后。与其他已研究的硬骨鱼不同,smyf5在体节发生之前就被检测到,但在轴旁细胞中未检测到。smyf5、smyoD1b和smyoG的表达域并不局限于s - smlc1表达区域,表明它们在快肌生成中发挥作用。从分节结束时起,每个MRF在新肌纤维产生区域或多或少都有表达,这些区域的前体细胞可能起源于单层s - smlc1(+)纤维外部表达Pax7的细胞层。在所研究的三个不同温度(2℃、5℃和8℃)下,尽管体节形成速度不同(在8℃、5℃和2℃时分别每5小时、8小时和15小时增加一个体节),smyoD1a和smyoG在体节阶段的表达模式相似。相比之下,与8℃相比,2℃时smyf5、sMRF4和s - smlc1的表达在体节阶段延迟,这可能导致影响后期肌肉表型的下游途径出现异时性。