牙鲆(Paralichthys olivaceus) Hsp90 mRNA在温度刺激和鳗弧菌感染下的表达特征

Expression Patterns of Hsp90 of Paralichthys olivaceus in Response to Temperature Treatment and Vibrio anguillarum Infection

中文关键词: 牙鲆  Hsp90α和Hsp90β  qPCR  温度刺激  鳗弧菌感染

英文关键词: Paralichthys olivaceus  Hsp90α and Hsp90β  qPCR  Temperature treatment  V. anguillarum infection

中文摘要:

      利用qPCR方法，检测Hsp90α和Hsp90β mRNA在不同月龄牙鲆(Paralichthys olivaceus)不同组织的表达水平，进一步检测在不同温度刺激、鳗弧菌感染后的表达变化。结果显示，Hsp90α和Hsp90β在不同发育期的13种组织(肝脏、脾脏、头肾、后肾、心、肌肉、背皮、胃、肠、鳃、腹鳍、脑和血液)中均有表达。Hsp90β表达水平整体高于Hsp90α；8月龄牙鲆在肝、鳃、背皮、背肌、腹鳍的Hsp90α表达量要显著高于其他发育期(1月龄、12月龄、24月龄)同组织的表达量(P<0.05)，而Hsp90β在不同月龄牙鲆的表达量也存在差异，在肝和肠的表达量较高，在鳃、背肌和腹鳍的表达量较低。将8月龄牙鲆在5–32℃刺激1 h，牙鲆肝、脾Hsp90α在高温(22–32℃)的表达量显著升高(P<0.05)，而Hsp90β在低温（5–10℃）的表达量显著升高(P<0.05)；在10℃、28℃连续刺激8 h，Hsp90α和Hsp90β的mRNA的表达量呈先上升后下降的趋势，Hsp90α的表达量变化幅度显著高于Hsp90β (P<0.05)。在鳗弧菌感染72 h内，牙鲆脾脏Hsp90α和Hsp90β的表达量先下降后上升，Hsp90β上升的幅度更加显著(P<0.05)。结果显示，Hsp90α和Hsp90β在不同月龄、不同组织的表达水平存在差异，Hsp90α在高温时对热应激的反应明显，Hsp90β在低温和对病原菌感染的反应明显，研究结果可为了解牙鲆Hsp90的作用机制提供参考。

英文摘要:

      In this study we investigated the expression patterns of Hsp90α and Hsp90β of Paralichthys olivaceus after the temperature treatment and Vibrio anguillarum infection. qRT-PCR was used to detect the expression of Hsp90 in different tissues of P. olivaceus during a series of growth stages (1-, 8-, 12-, and 24-month old). The results showed that Hsp90α and Hsp90β were expressed in 13 tissues including liver, spleen, head kidney, kidney, heart, muscle, dorsal skin, stomach, intestine, gill, pelvic fin, brain, and blood at all growth stages. The expression level of Hsp90β was overall higher than that of Hsp90α. Expression of Hsp90α was significantly higher at 8-month than at other growth stages in the liver, gills, dorsal skin, dorsal muscle, and pelvic. As the level of Hsp90β varied at different growth stages, it was observed to be higher in the liver and intestine. After a treatment at 5–32℃ for 1 h, the expression of Hsp90α in the liver and spleen was rapidly increased, whereas the expression of Hsp90β did not changed evidently. After a continuous treatment at 10℃ and 28℃ for 8 h, the expression of both Hsp90α and Hsp90β in the liver and spleen declined after an initial increase. Expression of Hsp90α varied in a wider range than Hsp90β. Within 72 h after the infection with V. anguillarum, expression of Hsp90α and Hsp90β in the spleen first rose and then decreased, and expression of Hsp90β had a larger variation range. These results indicated that Hsp90α and Hsp90β might have distinct expression patterns in different tissues at various growth stages. Hsp90α may be apt to respond to high temperature, while Hsp90β could be more sensitive to pathogen infection. These data will provide the basic data for better understanding of the functions of Hsp90 in P. olivaceus.