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底鳉对热应激的细胞反应:cDNA微阵列和蛋白质水平分析

The cellular response to heat stress in the goby Gillichthys mirabilis: a cDNA microarray and protein-level analysis.

作者信息

Buckley Bradley A, Gracey Andrew Y, Somero George N

机构信息

Hopkins Marine Station, Stanford University, Pacific Grove, CA 93950, USA.

出版信息

J Exp Biol. 2006 Jul;209(Pt 14):2660-77. doi: 10.1242/jeb.02292.

Abstract

The cellular response to stress relies on the rapid induction of genes encoding proteins involved in preventing and repairing macromolecular damage incurred as a consequence of environmental insult. To increase our understanding of the scope of this response, a cDNA microarray, consisting of 9207 cDNA clones, was used to monitor gene expression changes in the gill and white muscle tissues of a eurythermic fish, Gillichthys mirabilis (Gobiidae) exposed to ecologically relevant heat stress. In each tissue, the induction or repression of over 200 genes was observed. These genes are associated with numerous biological processes, including the maintenance of protein homeostasis, cell cycle control, cytoskeletal reorganization, metabolic regulation and signal transduction, among many others. In both tissues, the molecular chaperones, certain transcription factors and a set of additional genes with various functions were induced in a similar manner; however, the majority of genes displayed tissue-specific responses. In gill, thermal stress induced the expression of the major structural components of the cytoskeleton, whereas these same genes did not respond to heat in muscle. In muscle, many genes involved in promoting cell growth and proliferation were repressed, perhaps to conserve energy for repair and replacement of damaged macromolecules, but a similar repression was not observed in the gill. Many of the observed changes in gene expression were similar to those described in model species whereas many others were unexpected. Measurements of the concentrations of the protein products of selected genes revealed that in each case an induction in mRNA synthesis correlated with an increase in protein production, though the timing and magnitude of the increase in protein was not consistently predicted by mRNA concentration, an important consideration in assessing the condition of the stressed cell using transcriptomic analysis.

摘要

细胞对应激的反应依赖于迅速诱导编码蛋白质的基因,这些蛋白质参与预防和修复因环境损伤而产生的大分子损伤。为了增进我们对这种反应范围的理解,我们使用了一个由9207个cDNA克隆组成的cDNA微阵列,来监测广温性鱼类奇异吉利鲇(鲈形目)鳃和白肌组织在受到具有生态相关性的热应激时的基因表达变化。在每个组织中,观察到200多个基因的诱导或抑制。这些基因与众多生物学过程相关,包括蛋白质稳态的维持、细胞周期控制、细胞骨架重组、代谢调节和信号转导等等。在这两个组织中,分子伴侣、某些转录因子以及一组具有各种功能的其他基因以相似的方式被诱导;然而,大多数基因表现出组织特异性反应。在鳃中,热应激诱导了细胞骨架主要结构成分的表达,而这些相同的基因在肌肉中对热没有反应。在肌肉中,许多参与促进细胞生长和增殖的基因被抑制,这可能是为了节省能量用于修复和替换受损的大分子,但在鳃中未观察到类似的抑制现象。许多观察到的基因表达变化与在模式物种中描述的相似,而许多其他变化则出乎意料。对所选基因蛋白质产物浓度的测量表明,在每种情况下,mRNA合成的诱导与蛋白质产量的增加相关,尽管蛋白质增加的时间和幅度并不能始终由mRNA浓度预测,这是在使用转录组分析评估应激细胞状态时的一个重要考虑因素。

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