氨氮胁迫下钙离子稳态对凡纳滨对虾能量代谢和解毒代谢的影响

Effect of Ca2+ homeostasis on energy metabolism and detoxification metabolism in Litopenaeus vannamei exposed to ammonia stress

    摘要
  • 摘要: 为揭示钙离子 (Ca2+) 稳态在对虾应对氨氮耐受中的调控机制，本研究探究了氨氮胁迫下Ca2+ 对凡纳滨对虾 (Litopenaeus vannamei) 能量代谢和解毒代谢的影响。将对虾分为6组，分别注射相应的siRNA抑制Ca2+ 稳态调控基因SERCA (si-SERCA组)、MCU (si-MCU组)、IP3R (si-IP3R组) 的表达，以不注射任何物质 (NI组)、注射磷酸缓冲液 (PBS组)、注射非编码RNA (NC组) 作为对照。然后将其暴露于总氨氮质量浓度为11.3 mg·L−1的水体中，进行24 h的氨氮胁迫实验，测定能量代谢以及解毒代谢相关基因的表达水平。结果显示，si-SERCA、si-MCU和si-IP3R组能量代谢相关基因FAS、ACC、CPT-1、PK、PFK、HK的相对表达量显著高于对照组 (p<0.05)。si-SERCA组解毒代谢相关基因GS、GDH、GST的相对表达量在胁迫第6—第24 小时显著高于对照组 (p<0.05)。除胁迫第18小时外，si-MCU组GS基因的相对表达量高于对照组，而GDH基因的相对表达量在胁迫第6—第24 小时显著高于对照组，GST基因的相对表达量在胁迫第6—第18 小时显著高于对照组 (p<0.05)。si-IP3R组GS、GDH基因的相对表达量显著高于对照组，GST基因的相对表达量在胁迫第6—第24 小时显著高于对照组 (p<0.05)，且能量代谢相关基因的表达与解毒代谢相关基因的表达显著正相关 (p<0.05)。以上结果表明，在氨氮胁迫下，抑制Ca2+ 稳态相关基因的表达，对虾能量代谢以及解毒代谢水平显著升高，进而增强其氨氮耐受性。

     

    Abstract: To reveal the regulatory role of Ca2+ homeostasis in shrimp's ammonia nitrogen tolerance, we investigated the effects of ammonia stress on the energy metabolism and detoxification metabolism of Litopenaeus vannamei. We divided the shrimps into six groups, which were injected with specific siRNA to suppress the expression of Ca2+ stability regulation genes SERCA (si-SERCA group), MCU (si-MCU group), and IP3R (si-IP3R group), a group that received no substance (NI group), a group injected with phosphate-buffered saline (PBS group), and a group injected with non-coding RNA (NC group) as the control group. Then, we exposed them to a total ammonia concentration of 11.3 mg·L−1 for 24 h under ammonia stress conditions, and measured the expression levels of energy metabolism-related genes and detoxification metabolism-related genes. The results show that the expression levels of energy metabolism-related genes FAS, ACC, CPT-1, PK, PFK and HK in si-SERCA group, si-MCU group and si-IP3R group showed a significant increase compared with the control group (p<0.05). The expression levels of detoxification metabolism-related genes GS, GDH and GST in si-SERCA group were significantly higher than those in the control group at 6–24 h after stress (p<0.05). The expression level of GDH in si-MCU group was significantly higher than that in the control group at 6–24 h after stress, and the expression level of GST was significantly higher than that in the control group at 6–18 h after stress (p<0.05). The expression levels of GS, GDH and GST in si-IP3R group were significantly higher than those in the control group, and the expression level of GST was significantly higher than that in the control group at 6–24 h after stress (p<0.05). Furthermore, the expression of energy metabolism-related genes was significantly positively correlated with the expression of detoxification metabolism-related genes (p<0.05). The results indicate that under ammonia stress, inhibiting the expression of genes related to Ca2+ homeostasis significantly increases the energy metabolism and detoxification metabolism levels of shrimp, thereby enhancing their ammonia tolerance.

     

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