Molecular characterization and transcriptional regulation of the Na+/K+ ATPase α subunit isoforms during development and salinity challenge in a teleost fish, the Senegalese sole (Solea senegalensis)

• Published in: Comparative Biochemistry and Physiology - Part B: Biochemistry and Molecular Biology Vol. 175; pp. 23 - 38
• Main Authors: Armesto, Paula, Campinho, Marco A., Rodríguez-Rúa, Ana, Cousin, Xavier, Power, Deborah M., Manchado, Manuel, Infante, Carlos
• Format: Journal Article
• Language: English
• Published: England Elsevier Inc 01-09-2014; Elsevier
• Subjects: Amino Acid Sequence; Animal biology; Animals; Base Sequence; Development Biology; Flatfishes - growth & development; Flatfishes - metabolism; gene expression; Gills - metabolism; Larva - growth & development; Larva - metabolism; Life Sciences; Molecular Sequence Data; Na+,K+ ATPase; Organ Specificity; Osmoregulation; Osmotic Pressure; paralogous genes; Phylogeny; Protein Isoforms - genetics; Protein Isoforms - metabolism; Salinity; Senegalese sole; Sodium-Potassium-Exchanging ATPase - genetics; Sodium-Potassium-Exchanging ATPase - metabolism; aa; PBS; IHC; NKA; nt; ECD; Osmoregulation; kDa; TMD; DEPC; Senegalese sole; ppt; kb; cDNA; IC; SEM; paralogous genes; PK; DPH; Na+,K+ ATPase; gene expression; ISH; ML; K+ ATPase; Na+; osmoregulation; senegalese sole
• ISSN: 1096-4959; 1879-1107
• DOI: 10.1016/j.cbpb.2014.06.004

In the present work, five genes encoding different Na+,K+ ATPase (NKA) α-isoforms in the teleost Solea senegalensis are described for the first time. Sequence analysis of predicted polypeptides revealed a high degree of conservation across teleosts and mammals. Phylogenetic analysis clustered the five genes into three main clades: α1 (designated atp1a1a and atp1a1b), α2 (designated atp1a2) and α3 (designated atp1a3a and atp1a3b) isoforms. Transcriptional analysis in larvae showed distinct expression profiles during development. In juvenile tissues, the atp1a1a gene was highly expressed in osmoregulatory organs, atp1a2 in skeletal muscle, atp1a1b in brain and heart and atp1a3a and atp1a3b mainly in brain. Quantification of mRNA abundance after a salinity challenge showed that atp1a1a transcript levels increased significantly in the gill of soles transferred to high salinity water (60ppt). In contrast, atp1a3a transcripts increased at low salinity (5ppt). In situ hybridization (ISH) analysis revealed that the number of ionocytes expressing atp1a1a transcripts in the primary gill filaments was higher at 35 and 60ppt than at 5ppt and remained undetectable or at very low levels in the lamellae at 5 and 35ppt but increased at 60ppt. Immunohistochemistry showed a higher number of positive cells in the lamellae. Whole-mount analysis of atp1a1a mRNA in young sole larvae revealed that it was localized in gut, pronephric tubule, gill, otic vesicle, yolk sac ionocytes and chordacentrum. Moreover, atp1a1a mRNAs increased at mouth opening (3 DPH) in larvae incubated at 36ppt with a greater signal in gills.