Li Xuenan, Dai Xilin
Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai 201306, China; Shanghai Collaborative Innovation for Aquatic Animal Genetics and Breeding, Shanghai 201306, China; National Experimental Teaching Demonstration Centre for Aquatic Sciences, Shanghai Ocean University, Shanghai 201306, China.
Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai 201306, China; Shanghai Collaborative Innovation for Aquatic Animal Genetics and Breeding, Shanghai 201306, China; National Experimental Teaching Demonstration Centre for Aquatic Sciences, Shanghai Ocean University, Shanghai 201306, China.
Comp Biochem Physiol A Mol Integr Physiol. 2024 Dec;298:111749. doi: 10.1016/j.cbpa.2024.111749. Epub 2024 Sep 21.
The function of Litopenaeus vannamei Na/K/2Cl cotransporter 1 (NKCC1) under nitrite stress was investigated. The full-length cDNA sequence of the L. vannamei NKCC1 gene was cloned using the rapid amplification of cDNA ends (RACE) technique, and the sequence was analysed using bioinformatics tools. Expression and localisation of NKCC1 in tissues were assessed using real-time quantitative PCR and in situ hybridisation, respectively. The impact of nitrite stress on the survival, physiology, biochemistry and tissue structure of L. vannamei was investigated following silencing of NKCC1 by RNA interference. The 3143 bp cDNA sequence of L. vannamei NKCC1 encodes a polypeptide of 918 amino acids. It is evolutionarily conserved. NKCC1 expression was highest in gill tissue, particularly within cuticle and gill epithelial cells. After silencing NKCC1, an increase in shrimp survival was observed, accompanied by a significant reduction in nitrite entry into the body (P < 0.05). Moreover, the oxidative stress enzyme system remained unaffected and damage to gill tissue was alleviated. The results suggest that NKCC1 is involved in regulating nitrite uptake, and plays a crucial role in facilitating nitrite entry into the organism through gill tissue. The findings provide a vital experimental basis for addressing concerns related to nitrite toxicity.
研究了凡纳滨对虾钠/钾/2氯协同转运蛋白1（NKCC1）在亚硝酸盐胁迫下的功能。采用cDNA末端快速扩增（RACE）技术克隆凡纳滨对虾NKCC1基因的全长cDNA序列，并利用生物信息学工具对该序列进行分析。分别采用实时定量PCR和原位杂交技术评估NKCC1在组织中的表达和定位。通过RNA干扰沉默NKCC1后，研究亚硝酸盐胁迫对凡纳滨对虾存活、生理、生化和组织结构的影响。凡纳滨对虾NKCC1的3143 bp cDNA序列编码一个由918个氨基酸组成的多肽。它在进化上是保守的。NKCC1在鳃组织中的表达最高，尤其是在角质层和鳃上皮细胞内。沉默NKCC1后，观察到对虾存活率增加，同时进入体内的亚硝酸盐显著减少（P < 0.05）。此外，氧化应激酶系统未受影响，鳃组织的损伤得到缓解。结果表明，NKCC1参与调节亚硝酸盐的摄取，并在促进亚硝酸盐通过鳃组织进入生物体中起关键作用。这些发现为解决与亚硝酸盐毒性相关的问题提供了重要的实验依据。
