Abstract

Na+/K+-ATPase (NKA) belongs to the P-type ATPase family, whose members are located in the cell membrane and are distributed in diverse tissues and cells. The main function of the NKA is to regulate osmotic pressure. To better understand the role of NKA in osmoregulation, we first cloned and characterized the full-length cDNAs of NKA α subunit and β subunit from Pacific abalone Haliotis discus hannai in the current study. The predicted protein sequence of the NKA α subunit, as the catalytic subunit, was well conserved. In contrast, the protein sequence of the β subunit had low similarity with those of other species. Phylogenetic analysis revealed that both the α and β subunits of the NKA protein of Pacific abalone were clustered with those of the Gastropoda. Then, the relationship between salinity changes and the NKA was investigated. Sudden salinity changes (with low-salinity seawater (LSW) or high-salinity seawater (HSW)) led to clear changes in ion concentration (Na+ and K+) in hemolymph; however, the relative stability of ion concentrations in tissue revealed that Pacific abalone has a strong osmotic pressure regulation ability when faced with these salinity changes. Meanwhile, the expression and activity of the NKA was significantly decreased (in LSW group) or increased (in HSW group) during the ion concentration re-establishing stages, which was consistent with the coordinated regulation of ion concentration in hemolymph. Moreover, a positive correlation between cyclic adenosine monophosphate (cAMP) concentrations and NKA mRNA expression (NKA activity) was observed in mantle and gill. Therefore, the sudden salinity changes may affect NKA transcription activation, translation and enzyme activity via a cAMP-mediated pathway.

Highlights

  • The concentration of Na+ and K+ in LSW, HSW, and NSW were measured in order to test the differences in aquaculture waters with different salinities (Figure 2A). These data indicated that the ion concentration and salinity of seawater are positively correlated (P < 0.0001)

  • We found that the ion concentrations of Na+ and K+ in samples treated with LSW gradually decreased as the treatment time increased

  • To further explore the underlying molecular mechanism by which osmotic regulation leads to significant changes in HdNKA expression and activity after sudden treatment with different salinities, we examined the concentration of cyclic adenosine monophosphate (cAMP) in both mantle and gill (Figures 9A,B)

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Summary

Introduction

Abalones are large marine snails in the family Haliotidae and the genus Haliotis belonging to the class Gastropoda of the phylum Mollusca (Lv, 1978; Appeltans et al, 2014; Cook, 2016). Pacific abalone Haliotis discus hannai, which is naturally distributed in the Northwest Pacific, is considered an important commercially fishery industry animal in China. Temperature and salinity are the primary physical factors affecting the distribution and physiological metabolism of shellfish (Davenport et al, 1975; Burton, 1983; Navarro, 1988). Pacific abalone is a stenohaline species, and it is very sensitive to low salinity. The strain of P-97, which had experienced several generations of linearly selection since 1997, resulting in a significantly increased tolerance of temperature and salinity. The optimum growth salinity for the sixth generation of P-97 is approximately 24–36 part per thousand (ppt) (Kong et al, 2017)

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