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胰岛素家族生长因子对植入前小鼠胚胎的蛋白质合成具有特定作用。

Insulin family growth factors have specific effects on protein synthesis in preimplantation mouse embryos.

作者信息

Shi C Z, Collins H W, Buettger C W, Garside W T, Matschinsky F M, Heyner S

机构信息

Department of Obstetrics and Gynecology, University of Pennsylvania, Philadelphia.

出版信息

Mol Reprod Dev. 1994 Apr;37(4):398-406. doi: 10.1002/mrd.1080370406.

Abstract

Previously constructed protein databases for two stages of preimplantation mouse embryogenesis, the compacted eight-cell stage and the fully expanded blastocyst stage, have been used to analyze the effects of insulin, IGF-I, and IGF-II on protein synthesis in these developmental stages. Proteins were labeled by placing, for 2 hr, synchronous cohorts of 35-50 embryos into human tubal fluid (HTF) medium containing L-[35S]-methionine (1 mCi/ml) in the presence or absence of one of the growth factors. The embryos were then washed with medium and lysed. Samples were processed for 2-D gel analysis. For each embryonic stage and each growth factor, four or five experimental replicates were done and the gel images were compared using the PDQUEST system. Using the computer-assisted analysis, we were able to identify proteins that showed a statistically significant (P < 0.05) change in synthesis. At the eight-cell stage of development insulin caused increased synthesis of two proteins and decreased synthesis in three proteins. Insulin-treated blastocyst stage embryos exhibited an increased synthesis in eight proteins and decreased synthesis for one protein. The effect of IGF-I at the eight-cell stage of development was mostly inhibitory; the synthesis of only one protein increased and the synthesis of five proteins showed a decrease. Similar results were obtained with blastocyst stage embryos; four proteins demonstrated an increase in synthesis while 14 proteins showed a decrease. Eight-cell stage embryos incubated with IGF-II had seven proteins with a decreased synthesis, although in blastocyst stage embryos, nine proteins showed increased synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

先前构建的用于小鼠植入前胚胎发育两个阶段(致密化八细胞阶段和完全扩张囊胚阶段)的蛋白质数据库,已被用于分析胰岛素、胰岛素样生长因子-I(IGF-I)和胰岛素样生长因子-II(IGF-II)对这些发育阶段蛋白质合成的影响。通过将35 - 50个同步胚胎放入含有L-[35S]-甲硫氨酸(1 mCi/ml)的人输卵管液(HTF)培养基中2小时来标记蛋白质,培养基中存在或不存在其中一种生长因子。然后用培养基洗涤胚胎并裂解。对样品进行二维凝胶分析。对于每个胚胎阶段和每种生长因子,进行了四到五次实验重复,并使用PDQUEST系统比较凝胶图像。通过计算机辅助分析,我们能够鉴定出合成中显示出统计学显著变化(P < 0.05)的蛋白质。在八细胞发育阶段,胰岛素导致两种蛋白质的合成增加,三种蛋白质的合成减少。胰岛素处理的囊胚阶段胚胎表现出八种蛋白质的合成增加,一种蛋白质的合成减少。IGF-I在八细胞发育阶段的作用主要是抑制性的;只有一种蛋白质的合成增加,五种蛋白质的合成减少。囊胚阶段胚胎也得到了类似的结果;四种蛋白质的合成增加,而14种蛋白质的合成减少。用IGF-II孵育的八细胞阶段胚胎有七种蛋白质的合成减少,尽管在囊胚阶段胚胎中,有九种蛋白质的合成增加。(摘要截短于250字)

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