Meistertzheim Anne-Leila, Tanguy Arnaud, Moraga Dario, Thébault Marie-Thérèse
Laboratoire des Sciences de l'Environnement Marin, Institut Universitaire Européan de la Mer, Université de Bretagne occidentale, Plouzané, France.
FEBS J. 2007 Dec;274(24):6392-402. doi: 10.1111/j.1742-4658.2007.06156.x. Epub 2007 Nov 12.
Groups of oysters (Crassostrea gigas) were exposed to 25 degrees C for 24 days (controls to 13 degrees C) to explore the biochemical and molecular pathways affected by prolonged thermal stress. This temperature is 4 degrees C above the summer seawater temperature encountered in western Brittany, France where the animals were collected. Suppression subtractive hybridization was used to identify specific up- and downregulated genes in gill and mantle tissues after 7-10 and 24 days of exposure. The resulting libraries contain 858 different sequences that potentially represent highly expressed genes in thermally stressed oysters. Expression of 17 genes identified in these libraries was studied using real-time PCR in gills and mantle at different time points over the course of the thermal stress. Differential gene expression levels were much higher in gills than in the mantle, showing that gills are more sensitive to thermal stress. Expression of most transcripts (mainly heat shock proteins and genes involved in cellular homeostasis) showed a high and rapid increase at 3-7 days of exposure, followed by a decrease at 14 days, and a second, less-pronounced increase at 17-24 days. A slow-down in protein synthesis occurred after 24 days of thermal stress.
将几组牡蛎(太平洋牡蛎)置于25摄氏度环境中24天(对照组置于13摄氏度环境中),以探究长期热应激所影响的生化和分子途径。该温度比采集这些动物的法国布列塔尼西部夏季海水温度高4摄氏度。采用抑制性消减杂交技术,鉴定暴露7 - 10天和24天后鳃和外套膜组织中特异性上调和下调的基因。所得文库包含858个不同序列,这些序列可能代表热应激牡蛎中高表达的基因。在热应激过程中的不同时间点,使用实时PCR研究了这些文库中鉴定出的17个基因在鳃和外套膜中的表达情况。鳃中的差异基因表达水平远高于外套膜,表明鳃对热应激更敏感。大多数转录本(主要是热休克蛋白和参与细胞稳态的基因)的表达在暴露3 - 7天时呈现快速大幅增加,随后在14天时下降,在17 - 24天时出现第二次不太明显的增加。热应激24天后蛋白质合成减缓。