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Cell Stress and Chaperones

, Volume 24, Issue 3, pp 503–515 | Cite as

Characterization, functional analysis, and expression levels of three carbonic anhydrases in response to pH and saline–alkaline stresses in the ridgetail white prawn Exopalaemon carinicauda

  • Qianqian Ge
  • Jian LiEmail author
  • Jiajia Wang
  • Zhengdao Li
  • Jitao Li
Original Paper

Abstract

Carbonate alkalinity, salinity, and pH are three important stress factors for aquatic animals in saline–alkaline water. Carbonic anhydrases (CAs) catalyze the reversible reaction of CO2 reported to play an important role in the acid–base regulation in vertebrates. To explore the molecular mechanism of CAs efficacy in shrimp after their transfer into saline–alkaline water, the cDNAs of three CAs (EcCAc, EcCAg, and EcCAb) were cloned from Exopalaemon carinicauda. Sequence analysis showed that EcCAc and EcCAg both possessed a conserved α-CA domain and a proton acceptor site, and EcCAb contained a Pro-CA domain. Tissue expression analysis demonstrated that EcCAc and EcCAg were most abundantly in gills, and EcCAb was highly expressed in muscle. The cumulative mortalities remained below 25% under exposure to pH (pH 6 and pH 9), low salinity (5 ppt), or high carbonate alkalinity (5 and 10 mmol/L) after 72 h of exposure. However, mortalities increased up to 70% under extreme saline–alkaline stress (salinity 5 ppt, carbonate alkalinity 10 mmol/L, and pH 9) after 14 days of exposure. The EcCAc and EcCAg expressions in gills were significantly upregulated during the early period of pH and saline–alkaline stresses, while the EcCAb expressions showed no regular or large changes. The two-way ANOVA found significant interactions between salinity and carbonate alkalinity observed in EcCAc, EcCAg, and EcCAb expressions (p < 0.05). Furthermore, an RNA interference experiment resulted in increased mortality of EcCAc- and EcCAg-silenced prawns under saline–alkaline stress. EcCAc knockdown reduced expressions of Na+/H+ exchanger (EcNHE) and sodium bicarbonate cotransporter (EcNBC), and EcCAg knockdown reduced EcCAc, EcNHE, EcNBC, and V-type H+-ATPase (EcVTP) expressions. These results suggest EcCAc and EcCAg as important modulators in response to pH and saline–alkaline stresses in E. carinicauda.

Keywords

Carbonic anhydrase Exopalaemon carinicauda pH stress Saline–alkaline stress Expression analysis RNA interference 

Notes

Acknowledgements

This project was financially supported by the National Key R & D Program of China (2018YFD0901302), the National Natural Science Foundation of China (No. 31702319), Central Public-interest Scientific Institution Basal Research Fund, CAFS (NO.2019ZD0603), China Agriculture Research System-48 (CARS-48), The Program of Shandong Leading Talent (LNJY2015002).

Authors’ contributions

Qianqian Ge and Jian Li conceived and designed the experiments; Qianqian Ge, Jiajia Wang and Zhengdao Li performed the experiments; Qianqian Ge and Jiajia Wang analyzed the data; Jitao Li contributed materials; Qianqian Ge drafted the manuscript; and Jian Li made a critical revision of the manuscript.

Compliance with ethical standards

Conflicts of interest

The authors declare that they have no conflict of interest.

Supplementary material

12192_2019_987_MOESM1_ESM.docx (7.2 mb)
ESM 1 (DOCX 7354 kb)

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Copyright information

© Cell Stress Society International 2019

Authors and Affiliations

  • Qianqian Ge
    • 1
    • 2
  • Jian Li
    • 1
    • 2
    Email author
  • Jiajia Wang
    • 1
  • Zhengdao Li
    • 1
  • Jitao Li
    • 1
    • 2
  1. 1.Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research InstituteChinese Academy of Fishery SciencesQingdaoPeople’s Republic of China
  2. 2.Laboratory for Marine Fisheries Science and Food Production ProcessesQingdao National Laboratory for Marine Science and TechnologyQingdaoPeople’s Republic of China

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