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编码胰岛素和胰岛素样生长因子I受体的基因在非洲爪蟾卵母细胞和胚胎中表达。

Genes encoding receptors for insulin and insulin-like growth factor I are expressed in Xenopus oocytes and embryos.

作者信息

Scavo L, Shuldiner A R, Serrano J, Dashner R, Roth J, de Pablo F

机构信息

Diabetes Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892.

出版信息

Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6214-8. doi: 10.1073/pnas.88.14.6214.

Abstract

Insulin and insulin-like growth factor I (IGF-I) initiate their metabolic, growth, and differentiation effects through binding to the insulin receptor and the IGF-I receptor, two members of the tyrosine kinase family of receptors. To study the role of these peptides and receptors in early development, we used the polymerase chain reaction and embryo-derived RNA to generate partial cDNA sequences of the insulin receptor and IGF-I receptor from the amphibian Xenopus laevis. Three unique tyrosine kinase-related sequences were obtained. Two of the nucleotide sequences, XTK 1a and XTK 1b, corresponded to peptide that share 92% amino acid identity, and each is 89% identical to the human insulin receptor. The third sequence, XTK 2, corresponds to a peptide that has 92% amino acid identity with the human IGF-I receptor but only 80% identity with XTK 1a and XTK 1b. On the basis of these similarities, the pattern of conserved amino acids, and the tetraploid nature of the Xenopus genome, we suggest that XTK 1a and XTK 1b most likely represent the product of two different nonallelic insulin receptor genes, while XTK 2 may be one of the probable two Xenopus IGF-I receptor genes. By reverse transcription-polymerase chain reaction and gene-specific hybridization, expression of the three XTK sequences was detected in the oocyte, unfertilized egg, and embryos through gastrulation, neurulation, and tailbud stages. Competition binding assays with Xenopus membrane preparations demonstrated insulin receptors and IGF-I receptors in older tadpoles. IGF-I receptors were also present in oocytes, eggs, and gastrula embryos. By contrast, insulin binding was present but atypical in oocytes and was barely detected in eggs and gastrula embryos. The expression of receptors for insulin and IGF-I in early Xenopus embryos and their apparent distinct developmental regulation suggest that these molecules and their ligands may be important in early Xenopus development.

摘要

胰岛素和胰岛素样生长因子I(IGF-I)通过与胰岛素受体和IGF-I受体结合来启动它们的代谢、生长和分化作用,这两种受体是酪氨酸激酶家族受体的成员。为了研究这些肽和受体在早期发育中的作用,我们使用聚合酶链反应和胚胎来源的RNA从两栖动物非洲爪蟾中生成胰岛素受体和IGF-I受体的部分cDNA序列。获得了三个独特的酪氨酸激酶相关序列。其中两个核苷酸序列,XTK 1a和XTK 1b,对应于氨基酸同一性为92%的肽,并且每个与人类胰岛素受体的同一性为89%。第三个序列,XTK 2,对应于一种与人类IGF-I受体具有92%氨基酸同一性但与XTK 1a和XTK 1b仅具有80%同一性的肽。基于这些相似性、保守氨基酸模式以及爪蟾基因组的四倍体性质,我们认为XTK 1a和XTK 1b最有可能代表两个不同的非等位胰岛素受体基因的产物,而XTK 2可能是爪蟾两个可能的IGF-I受体基因之一。通过逆转录-聚合酶链反应和基因特异性杂交,在卵母细胞、未受精卵以及通过原肠胚形成、神经胚形成和尾芽阶段的胚胎中检测到了这三个XTK序列的表达。用爪蟾膜制剂进行的竞争结合试验证明了在较老的蝌蚪中有胰岛素受体和IGF-I受体。IGF-I受体也存在于卵母细胞、卵和原肠胚胚胎中。相比之下,胰岛素结合在卵母细胞中存在但不典型,在卵和原肠胚胚胎中几乎检测不到。胰岛素和IGF-I受体在早期爪蟾胚胎中的表达及其明显不同的发育调控表明,这些分子及其配体可能在爪蟾早期发育中很重要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/630b/52053/b55b47992df5/pnas01064-0287-a.jpg

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