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利用新型 RNA-Seq 程序对珊瑚幼虫(Acropora millepora)在高温和附着诱导物下的基因表达反应进行分析。

Profiling gene expression responses of coral larvae (Acropora millepora) to elevated temperature and settlement inducers using a novel RNA-Seq procedure.

机构信息

Integrative Biology Section, The University of Texas at Austin, Austin, TX 78712, USA.

出版信息

Mol Ecol. 2011 Sep;20(17):3599-616. doi: 10.1111/j.1365-294X.2011.05205.x. Epub 2011 Jul 29.

Abstract

Elevated temperatures resulting from climate change pose a clear threat to reef-building corals; however, the traits that might influence corals' survival and dispersal during climate change remain poorly understood. Global gene expression profiling is a powerful hypothesis-forming tool that can help elucidate these traits. Here, we applied a novel RNA-Seq protocol to study molecular responses to heat and settlement inducers in aposymbiotic larvae of the reef-building coral Acropora millepora. This analysis of a single full-sibling family revealed contrasting responses between short- (4-h) and long-term (5-day) exposures to elevated temperatures. Heat shock proteins were up-regulated only in the short-term treatment, while the long-term treatment induced the down-regulation of ribosomal proteins and up-regulation of genes associated with ion transport and metabolism (Ca(2+) and CO(3)(2-)). We also profiled responses to settlement cues using a natural cue (crustose coralline algae, CCA) and a synthetic neuropeptide (GLW-amide). Both cues resulted in metamorphosis, accompanied by differential expression of genes with known developmental roles. Some genes were regulated only by the natural cue, which may correspond to the recruitment-associated behaviour and morphology changes that precede metamorphosis under CCA treatment, but are bypassed under GLW-amide treatment. Validation of these expression profiles using qPCR confirmed the quantitative accuracy of our RNA-Seq approach. Importantly, qPCR analysis of different larval families revealed extensive variation in these responses depending on genetic background, including qualitative differences (i.e. up-regulation in one family and down-regulation in another). Future studies of gene expression in corals will have to address this genetic variation, which could have important adaptive consequences for corals during global climate change.

摘要

气候变化导致的温度升高对造礁珊瑚构成了明显威胁;然而,影响珊瑚在气候变化过程中生存和扩散的特征仍知之甚少。全局基因表达谱分析是一种强大的假设形成工具,可以帮助阐明这些特征。在这里,我们应用一种新的 RNA-Seq 方案来研究无共生幼虫对热和定居诱导物的分子反应,这些幼虫来自造礁珊瑚 Acropora millepora。对一个单一家系的分析显示,短期(4 小时)和长期(5 天)暴露于高温下的反应截然不同。热休克蛋白仅在短期处理中上调,而长期处理则诱导核糖体蛋白下调和与离子转运和代谢(Ca(2+) 和 CO(3)(2-))相关的基因上调。我们还使用天然诱导物(结皮层珊瑚藻,CCA)和合成神经肽(GLW-酰胺)来分析对定居诱导物的反应。这两种诱导物都导致了变态,伴随着已知发育作用的基因的差异表达。一些基因仅受天然诱导物调节,这可能对应于在 CCA 处理下,先于变态的与招募相关的行为和形态变化,但在 GLW-酰胺处理下被绕过。使用 qPCR 对这些表达谱进行验证,证实了我们的 RNA-Seq 方法的定量准确性。重要的是,对不同幼虫家系的 qPCR 分析显示,这些反应存在广泛的遗传背景差异,包括定性差异(即一个家系上调,另一个家系下调)。未来对珊瑚基因表达的研究必须解决这种遗传变异,这可能对珊瑚在全球气候变化过程中具有重要的适应性影响。

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