Makabe Kazuhiro W, Fujiwara Shigeki, Saiga Hidetoshi, Satoh Noriyuki
Department of Zoology, Kyoto University, 606, Kyoto, Japan.
Department of Molecular Biology, School of Medicine, University of Occupational and Environmental Health, 807, Kitakyushu, Japan.
Rouxs Arch Dev Biol. 1990 May;199(5):307-313. doi: 10.1007/BF01709509.
In ascidians the lineage of embryonic precursor cells is well documented. In this study, we investigated temporal and spatial expression of myosin heavy chain (MHC) gene during embryogenesis of the ascidianHalocynthia roretzi. When the occurrence of MHC transcripts was examined by Northern blot hybridization and in situ hybridization with specific cDNA and antisense RNA probes, transcripts were undetectable in fertilized eggs and cleavage-stage embryos. This suggests that the maternal message for MHC is not directly associated with the synthesis of MHC protein in ascidian embryos. MHC transcripts were initially observed at the gastrula stage. In situ hybridization of whole-mount preparations demonstrated that the transcripts first appeared in nuclei of primary-lineage muscle cells of the early-to-middle gastrula and accumulated rapidly as development progressed. The occurrence of MHC transcripts was restricted to differentiating muscle cells. No hybridization signal was detected in mesenchyme cells which are thought to form adult body-wall muscle. After metamorphosis the amount of MHC transcripts decreased; they became undetectable in juveniles about 10 days after metamorphosis, suggesting an intermission of MHC gene activity prior to the formation of adult bodywall muscle. Thus the expression of MHC gene in ascidian embryos was strictly regulated in both spatial and temporal orders and occurred only in muscle lineage cells.
在海鞘中,胚胎前体细胞的谱系有详尽记录。在本研究中,我们调查了海鞘罗津海鞘胚胎发育过程中肌球蛋白重链(MHC)基因的时空表达。当通过Northern印迹杂交以及用特异性cDNA和反义RNA探针进行原位杂交来检测MHC转录本的出现情况时,在受精卵和卵裂期胚胎中未检测到转录本。这表明在海鞘胚胎中,MHC的母体信息与MHC蛋白的合成没有直接关联。MHC转录本最初在原肠胚阶段被观察到。整装标本的原位杂交表明,转录本首先出现在早至中晚期原肠胚的初级谱系肌肉细胞的细胞核中,并随着发育进程迅速积累。MHC转录本的出现仅限于正在分化的肌肉细胞。在被认为会形成成体体壁肌肉的间充质细胞中未检测到杂交信号。变态后,MHC转录本的数量减少;在变态后约10天的幼体中它们变得无法检测到,这表明在成体体壁肌肉形成之前MHC基因活性有一个间断期。因此,海鞘胚胎中MHC基因的表达在时空顺序上受到严格调控,并且仅在肌肉谱系细胞中发生。