Shuldiner A R, de Pablo F, Moore C A, Roth J
Section on Receptors and Hormone Action, National Institute of Diabetes, and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7679-83. doi: 10.1073/pnas.88.17.7679.
Insulin, traditionally regarded as a metabolic hormone, also can potently stimulate growth and differentiation in many cell types. To study further the potential role of insulin during early embryogenesis, we have used the amphibian Xenopus laevis, a versatile model of vertebrate development. Using (i) nucleotide sequences of two previously cloned cDNAs that correspond to two different nonallelic Xenopus insulin genes (both of which are expressed in the adult pancreas) and (ii) a modification of the highly sensitive reverse transcription-polymerase chain reaction (RT-PCR) method developed in our laboratory, designated RNA template-specific PCR (RS-PCR), we now find that mRNAs for both Xenopus insulins I and II are present in mature (stage VI) oocytes but not in less-mature oocytes (stages I and IV) or in unfertilized eggs. The Xenopus insulin II gene is differentially expressed during early neurulation (stage 13), while only the insulin I gene is expressed at stage 21, when the neural tube is closing and cephalization is beginning. During later stages (i.e., stage 26) there is a region in the head that appears to be transcribing only the insulin I gene, while mRNAs for both insulins I and II are present in the body region. These findings show that the two nonallelic insulin genes are expressed differentially in Xenopus embryos in a stage- and region-specific manner; because appropriate receptors are also present, we suggest a role for insulin during early nervous system development well before the emergence of pancreatic beta cells.
胰岛素传统上被视为一种代谢激素,但它也能有力地刺激多种细胞类型的生长和分化。为了进一步研究胰岛素在早期胚胎发育过程中的潜在作用,我们使用了非洲爪蟾(Xenopus laevis),这是一种用于脊椎动物发育研究的通用模型。利用(i)两个先前克隆的cDNA的核苷酸序列,它们对应于非洲爪蟾两个不同的非等位胰岛素基因(这两个基因在成年胰腺中均有表达),以及(ii)对我们实验室开发的高度灵敏的逆转录 - 聚合酶链反应(RT-PCR)方法的一种改进,即RNA模板特异性PCR(RS-PCR),我们现在发现非洲爪蟾胰岛素I和II的mRNA存在于成熟(VI期)卵母细胞中,但不存在于不太成熟的卵母细胞(I期和IV期)或未受精卵中。非洲爪蟾胰岛素II基因在早期神经胚形成阶段(13期)差异表达,而在21期,即神经管闭合和头部开始形成时,只有胰岛素I基因表达。在后期阶段(即26期),头部有一个区域似乎只转录胰岛素I基因,而胰岛素I和II的mRNA都存在于身体区域。这些发现表明两个非等位胰岛素基因在非洲爪蟾胚胎中以阶段和区域特异性的方式差异表达;由于相应的受体也存在,我们认为在胰腺β细胞出现之前很久,胰岛素就在早期神经系统发育中发挥作用。