采用实时荧光定量RT-PCR方法,检测了注射溶藻弧菌(Vibiro alginolyticus)后近江牡蛎鳃,闭壳肌,消化腺,外套膜,心脏以及血细胞中HSPTO基因的表达变化。结果显示近江牡蛎这五种器官组织中的HSPTO基因表达量均出现显著性高表达,且在鳃、外套膜和血细胞中的HSPTO基因表达变化规律表现为典型的时间依赖性。血细胞中,显著高表达的峰值出现在24h,至72h恢复到对照水平,高表达持续时间最长;鳃中表达峰值出现时间较早,在第3h,随后在第12h便恢复到对照水平;外套膜,消化腺以及心脏中的峰值分别出现在6h,6h和3h,而在闭壳肌组织中,没出现显著性高表达。由此可见,近江牡蛎HSP70s可能在机体抗菌免疫过程中起了重要作用。
In order to investigate the role of HSPTO of oyster (Crassostrea hongkongensis) against bacterial infection, we established a real-time, quantitative fluorescent RT-PCR method to examine the expression patterns of HSP70 mRNA in mantle, digestive gland, gills, muscle, heart and haemocytes of the oyster responding to Vibiro alginolyticus challenge. The results showed that after the oysters were injected with live V. alginolyticus, HSP70 mRNA expression in gills, mantle and haemocytes was up-regulated at first, peaked at 3 hr, 6 hr and 24 hr, respectively, then gradually decreased to the control level at 12 hr, 24 hr sand 72 hr, respectively. The phenomenon was defined as time-dependent expression pattern of HSP70 gene. Meanwhile, the HSP70 mRNA levels in heart and digestive gland significantly increased and reached the highest values at 3 hr and 6 hr, respectively, and then dropped rapidly. But in muscle, the HSP70 mRNA expression did not increased upon the bacterial challenge during the entire experiment period. The up-regulated mRNA expression of HSP70 induced by bacterial challenge suggested an important role for HSP70 gene in immune response in oysters.