Nishikata T, Mita-Miyazawa I, Deno T, Satoh N
Department of Zoology, Kyoto University, Japan.
Development. 1987 Feb;99(2):163-71. doi: 10.1242/dev.99.2.163.
Utilizing a muscle-specific monoclonal antibody (Mu-2) as a probe, we analysed developmental mechanisms involved in muscle cell differentiation in ascidian embryos. The antigen recognized by Mu-2 was a single polypeptide with a relative molecular mass of about 220 X 10(3). It first appeared at the early tailbud stage and continued to be expressed until the swimming larva stage. There were distinct and separate puromycin and actinomycin D sensitivity periods during the occurrence of the antigen, suggesting the new synthesis of the polypeptide by developing muscle cells. Embryos that had been permanently arrested with aphidicolin in the early cleavage stages up to the 32-cell stage did not express the antigen. DNA replications may be required for the antigen expression. Embryos that had been arrested with cytochalasin B in the 8-cell and later stages developed the antigen, and the number and position of the arrested blastomeres exhibiting the differentiation marker almost corresponded to those of the B4.1-line muscle lineage. Furthermore, in quarter embryos developed from each blastomere pair isolated from the 8-cell embryo, all the B4.1 as well as a part of b4.2 partial embryos expressed the antigen, while the a4.2 and A4.1 partial embryos did not show the antigen expression. These results may provide further support for the existence of cytoplasmic determinants for muscle cell differentiation in this mosaic egg.
利用一种肌肉特异性单克隆抗体(Mu-2)作为探针,我们分析了海鞘胚胎中肌肉细胞分化所涉及的发育机制。Mu-2识别的抗原是一种相对分子质量约为220×10³的单一多肽。它最早出现在早期尾芽阶段,并持续表达至游泳幼虫阶段。在抗原出现过程中存在明显且独立的嘌呤霉素和放线菌素D敏感时期,这表明发育中的肌肉细胞会重新合成该多肽。在早期卵裂阶段直至32细胞阶段被阿非迪霉素永久阻滞的胚胎不表达该抗原。抗原表达可能需要DNA复制。在8细胞及后续阶段被细胞松弛素B阻滞的胚胎会发育出该抗原,显示出分化标记的被阻滞卵裂球的数量和位置几乎与B4.1系肌肉谱系的一致。此外,从8细胞胚胎分离出的每个卵裂球对发育而成的四分之一胚胎中,所有B4.1以及部分b4.2四分之一胚胎表达了该抗原,而a4.2和A4.1四分之一胚胎未显示抗原表达。这些结果可能为这种镶嵌卵中肌肉细胞分化的细胞质决定因素的存在提供进一步支持。