Heideveld M, Ayoubi T A, van de Wiel M H, Martens G J, Durston A J
Hubrecht Laboratory, Netherlands Institute for Developmental Biology, Utrecht.
Differentiation. 1993 Mar;52(3):195-200. doi: 10.1111/j.1432-0436.1993.tb00631.x.
Proopomelanocortin (POMC) is the precursor protein for a number of peptide hormones and neuropeptides, and the POMC gene is transcriptionally very active in the pars intermedia of the pituitary of the amphibian Xenopus laevis (Xenopus). We analysed the expression of this gene during Xenopus embryogenesis, in order to examine whether it can function as a (novel) neural marker. We investigated the spatio-temporal distribution of POMC mRNA, using a single-stranded probe that corresponds to the 3' untranslated region of Xenopus POMC gene B mRNA. Gene transcripts were first detected at stage 25 of development via RNase protection assays. In situ hybridization analysis performed at stage 46 showed clearly that these transcripts are localised in a region representing the future pars intermedia of the pituitary. Experiments using Xenopus explants indicate that the POMC gene can be used successfully as an indirect marker in studies on neural induction: in the absence of interactions with mesoderm, ectoderm fails to express the POMC gene, whereas POMC transcripts are readily detectable in conjugates of ectoderm and mesoderm. Artificial application of two different signals, which are likely to be relevant for neural differentiation (namely retinoic acid and the activation of protein kinase C via phorbol ester), was not effective in evoking POMC gene expression in cultured ectoderm explants. However, retinoic acid treatment of conjugates of Xenopus ectoderm and mesoderm successfully prevented POMC expression. We conclude that POMC gene expression can be used as an indirect marker for anterior neural differentiation in Xenopus.
促阿黑皮素原(POMC)是多种肽类激素和神经肽的前体蛋白,POMC基因在两栖动物非洲爪蟾(爪蟾)垂体中间部的转录活性非常高。我们分析了该基因在爪蟾胚胎发育过程中的表达情况,以检验它是否能作为一种(新型)神经标志物发挥作用。我们使用与爪蟾POMC基因B mRNA的3'非翻译区对应的单链探针,研究了POMC mRNA的时空分布。通过核糖核酸酶保护分析在发育的第25阶段首次检测到基因转录本。在第46阶段进行的原位杂交分析清楚地表明,这些转录本定位于代表垂体未来中间部的区域。使用爪蟾外植体的实验表明,POMC基因可成功用作神经诱导研究中的间接标志物:在没有与中胚层相互作用的情况下,外胚层无法表达POMC基因,而在外胚层和中胚层的结合物中很容易检测到POMC转录本。人工施加两种可能与神经分化相关的不同信号(即视黄酸和通过佛波酯激活蛋白激酶C),在诱导培养的外胚层外植体中POMC基因表达方面无效。然而,视黄酸处理爪蟾外胚层和中胚层的结合物成功地阻止了POMC表达。我们得出结论,POMC基因表达可作为爪蟾前神经分化的间接标志物。