Apoptosis-stimulating protein of p53 (ASPP) participates in the regulation of apoptosis in Litopenaeus vannamei under ammonia-N and nitrite-N stress

IF 3.2 2区农林科学 Q1 FISHERIES

Aquaculture Reports Pub Date : 2024-10-09 DOI:10.1016/j.aqrep.2024.102413

Xiaoxun Zhou, Hongbiao Zhuo, Lanting Lin, Yuan Zhang, Jinyan Li, Shuo Fu, Guangbo Wu, Chaoan Guo, Jianyong Liu

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引用次数: 0

Abstract

Apoptosis-stimulating protein of p53 (ASPP) is a key regulatory factor closely related to p53 in apoptosis pathway. To further investigate the molecular mechanisms of ASPP in Litopenaeus vannamei, the expression of LvASPP mRNA under ammonia-N and nitrite-N stress was explored, and the effects of knocking out LvASPP on mortality, histological damage, and the apoptosis pathway in L. vannamei under ammonia-N and nitrite-N stress were investigated. Healthy L. vannamei (7.78 ± 0.70 g) were used in this study. After shrimp were stressed with an ammonia-N concentration of 30.00 mg/L for 48 h, qRT-PCR was used to detect a significant increase in LvASPP mRNA expression in the hepatopancreas, gills, and muscle. Following 48 h of nitrite-N stress at a concentration of 60.00 mg/L, LvASPP mRNA expression was significantly increased in the hepatopancreas. The survival rate notably increased under 80 h of ammonia-N stress (25.00 mg/L) after LvASPP RNA interference (30 % more than the control group), and the number of shrimp deaths decreased after 48 h of nitrite-N stress. Moreover, under the stress of ammonia-N and nitrite-N respectively, LvASPP silencing reduced the expression of p53, and led to a decrease in the expression of apoptosis-related genes (Bax, Apaf-1, Caspase 9, MDM2). Caspase 3 activity, TUNEL-positive cells and the apoptotic index in the hepatopancreas markedly reduced under ammonia-N and nitrite-N stress. The potential pathway suggests that inhibiting LvASPP reduces the mRNA expression of p53, which leads to a decrease in Caspase 3 activity, inhibiting apoptosis in the hepatopancreatic cells of L. vannamei under ammonia-N and nitrite-N stress. These data indicate that the knockdown of LvASPP positively impacted the tolerance of L. vannamei to ammonia-N and nitrite-N stress by regulating the apoptosis pathway. This suggests that employing gene-targeted dsRNA could be an effective strategy for alleviating the environmental pressures faced by shrimp in aquaculture management.

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p53凋亡刺激蛋白（ASPP）参与调控氨-N和亚硝酸盐-N胁迫下的万年青凋亡

p53凋亡刺激蛋白（ASPP）是凋亡通路中与p53密切相关的关键调控因子。为了进一步研究ASPP在凡纳滨对虾中的分子机制，本研究探讨了LvASPP mRNA在氨-N和亚硝-N胁迫下的表达，并研究了敲除LvASPP对凡纳滨对虾在氨-N和亚硝-N胁迫下的死亡率、组织学损伤和细胞凋亡途径的影响。本研究使用了健康的凡纳滨对虾（7.78 ± 0.70 g）。用浓度为 30.00 mg/L 的氨氮对对虾胁迫 48 小时后，使用 qRT-PCR 检测 LvASPP mRNA 在肝胰脏、鳃和肌肉中的表达。在浓度为 60.00 mg/L 的亚硝酸盐-N 胁迫 48 小时后，LvASPP mRNA 在肝胰脏中的表达明显增加。LvASPP RNA干扰后，对虾在氨氮胁迫（25.00 mg/L）80小时后的存活率明显提高（比对照组高出30%），亚硝酸盐-N胁迫48小时后对虾死亡数量减少。此外，在氨氮和亚硝酸盐-N胁迫下，LvASPP沉默可降低p53的表达，并导致凋亡相关基因（Bax、Apaf-1、Caspase 9、MDM2）表达的减少。在氨-N和亚硝-N胁迫下，肝胰腺中的Caspase 3活性、TUNEL阳性细胞和凋亡指数明显降低。潜在的途径表明，抑制 LvASPP 可降低 p53 的 mRNA 表达，从而降低 Caspase 3 的活性，抑制氨-N 和亚硝-N 胁迫下的凡纳滨鲤肝胰腺细胞凋亡。这些数据表明，敲除 LvASPP 可通过调节细胞凋亡途径，对凡纳滨鲤对氨氮和亚硝氮胁迫的耐受性产生积极影响。这表明，在水产养殖管理中，采用基因靶向 dsRNA 可能是缓解对虾面临的环境压力的有效策略。

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来源期刊

Aquaculture Reports Agricultural and Biological Sciences-Animal Science and Zoology

CiteScore

5.90

自引率

8.10%

发文量

469

审稿时长

77 days

期刊介绍： Aquaculture Reports will publish original research papers and reviews documenting outstanding science with a regional context and focus, answering the need for high quality information on novel species, systems and regions in emerging areas of aquaculture research and development, such as integrated multi-trophic aquaculture, urban aquaculture, ornamental, unfed aquaculture, offshore aquaculture and others. Papers having industry research as priority and encompassing product development research or current industry practice are encouraged.