Skip to main content
SpringerLink
Log in

Differential expression of Na+, K+-ATPase α-1 isoforms during seawater acclimation in the amphidromous galaxiid fish Galaxias maculatus

  • Original Paper
  • Published:
Journal of Comparative Physiology B Aims and scope Submit manuscript

Abstract

Inanga (Galaxias maculatus) is an amphidromous fish with a well-known capacity to withstand a wide range of environmental salinities. To investigate the molecular mechanisms facilitating acclimation of inanga to seawater, several isoforms of the Na+, K+-ATPase ion transporter were identified. This included three α-1 (a, b and c), an α-2 and two α-3 (a and b) isoforms. Phylogenetic analysis showed that the inanga α-1a and α-1b formed a clade with the α-1a and α-1b isoforms of rainbow trout, while another clade contained the α-1c isoforms of these species. The expression of all the α-1 isoforms was modulated after seawater exposure (28 ‰). In gills, the expression of the α-1a isoform was progressively down-regulated after seawater exposure, while the expression of the α-1b isoform was up-regulated. The α-1c isoform behaved similarly to the α-1a, although changes were less dramatic. Physiological indicators of salinity acclimation matched the time frame of the changes observed at the molecular level. A 24-h osmotic shock period was highlighted by small increases in plasma osmolality, plasma Na+ and a decrease in muscle tissue water content. Thereafter, these values returned close to their pre-exposure (freshwater) values. Na+, K+-ATPase activity showed a decreasing trend over the first 72 h following seawater exposure, but activity increased after 240 h. Our results indicate that inanga is an excellent osmoregulator, an ability that is conferred by the rapid activation of physiological and molecular responses to salinity change.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Blanco G, Mercer RW (1998) Isozymes of the Na-K-ATPase: heterogeneity in structure, diversity in function. Am J Physiol 275:F633–F650

    PubMed  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Bystriansky JS, Richards JG, Schulte PM, Ballantyne JS (2006) Reciprocal expression of gill Na+/K+-ATPase α-subunit isoforms α-1a and α-1b during seawater acclimation of three salmonid fishes that vary in their salinity tolerance. J Exp Biol 209:1848–1858

    Article  PubMed  CAS  Google Scholar 

  • Chessman BC, Williams WD (1975) Salinity tolerance and osmoregulatory ability of Galaxias maculatus (Jenyns) (Pisces, Salmoniformes, Galaxiidae). Freshwat Biol 5:135–140

    Article  Google Scholar 

  • Cutler CP, Sanders IL, Hazon N, Cramb G (1995a) Primary sequence, tissue specificity and expression of the Na+, K+-ATPase α-1 subunit in the European eel (Anguilla anguilla). Comp Biochem Physiol 111B:567–573

    CAS  Google Scholar 

  • Cutler CP, Sanders IL, Hazon N, Cramb G (1995b) Primary sequence, tissue-specificity and messenger-RNA expression of the Na+, K+-ATPase α-1 subunit in the European eel (Anguilla anguilla). Fish Physiol Biochem 14:423–429

    Article  CAS  Google Scholar 

  • D’Cotta H, Valotaire C, Le Gac F, Prunet P (2000) Synthesis of gill Na+-K+-ATPase in Atlantic salmon smolts: differences in α-mRNA and α-protein levels. Am J Physiol 278:R101–R110

    Google Scholar 

  • Evans DH, Piermarini PM, Choe KP (2005) The multifunctional fish gill: dominant site of gas exchange, osmoregulation, acid-base regulation, and excretion of nitrogenous waste. Physiol Rev 85:97–177

    Article  PubMed  CAS  Google Scholar 

  • Feng SH, Leu JH, Yang CH, Fang MJ, Huang CJ, Hwang PP (2002) Gene expression of Na+-K+-ATPase alpha 1 and alpha 3 subunits in gills of the teleost Oreochromis mossambicus, adapted to different environmental salinities. Mar Biotechnol 4:379–391

    Article  PubMed  CAS  Google Scholar 

  • Guynn SR, Scofield MA, Petzel DH (2002) Identification of mRNA and protein expression of the Na/K-ATPase α-1, α-2 and α-3 subunit isoforms in Antarctic and New Zealand nototheniid fishes. J Exp Mar Biol Eco 273:15–32

    Article  CAS  Google Scholar 

  • Hwang PP, Sun CM, Wu SM (1998) Changes of plasma osmolarity, chloride concentration and gill Na-K-ATPase activity in tilapia Oreochromis mossambicus, during seawater acclimation. Mar Biol 100:295–299

    Article  Google Scholar 

  • Johnston CE, Saunders RL (1981) Parr-smolt transformation of yearling Atlantic salmon (Salmo salar) at several rearing temperatures. Can J Fish Aquat Sci 38:1189–1198

    Article  Google Scholar 

  • Jorgensen PL (2008) Importance for absorption of Na+ from freshwater of lysine, valine and serine substitutions in the α-1a isoform of Na, K–ATPase in the gills of rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar). J Membr Biol 223:37–47

    Article  PubMed  CAS  Google Scholar 

  • Kelly SP, Woo NYS (1999) Cellular and biochemical characterization of hyposmotic adaptation in a marine teleost, Sparus sarba. Zool Sci 16:505–514

    Article  CAS  Google Scholar 

  • Lin CH, Tsai RS, Lee TH (2004) Expression and distribution of Na, K-ATPase in gill and kidney of the spotted green pufferfish, Tetraodon nigroviridis, in response to salinity challenge. Comp Biochem Physiol 138A:287–295

    CAS  Google Scholar 

  • Mackie P, Wright PA, Glebe BD, Ballantyne JS (2005) Osmoregulation and gene expression of Na+/K+-ATPase in families of Atlantic salmon (Salmo salar) smolts. Can J Fish Aquat Sci 62:2661–2672

    Article  CAS  Google Scholar 

  • Madsen SS, Naamansen ET (1989) Plasma ionic regulation and gill Na+, K+-ATPase changes during rapid transfer to seawater of yearling rainbow trout Salmo gairdneri: time course and seasonal variation. J Fish Biol 34:829–840

    Article  CAS  Google Scholar 

  • Madsen SS, Jensen MK, Nohr J, Kristiansen K (1995) Expression of Na+, K+-ATPase in the brown trout Salmo trutta: in vivo modulation by hormones and seawater. Am J Physiol-Regul Integr Comp Physiol 269:R1339–R1345

    CAS  Google Scholar 

  • Madsen SS, Kiilerich P, Tipsmark CK (2009) Multiplicity of expression of Na+, K+-ATPase α-subunit isoforms in the gill of Atlantic salmon (Salmo salar): cellular localisation and absolute quantification in response to salinity change. J Exp Biol 212:78–88

    Article  PubMed  CAS  Google Scholar 

  • Marshall WS, Grosell M (2005) Ion transport, osmoregulation and acid-base balance. In: Evans DH, Claiborne JB (eds) The physiology of fishes. CRC Press, Boca Raton

    Google Scholar 

  • Martínez-Álvarez RM, Hidalgo MC, Domezain A, Morales AE, García-Gallego M, Sanz A (2002) Physiological changes of sturgeon Acipenser naccarii caused by increasing environmental salinity. J Exp Biol 205:3699–3706

    PubMed  Google Scholar 

  • McCormick SD (1993) Methods for nonlethal gill biopsy and measurement of Na+, K+-ATPase activity. Can J Fish Aquat Sci 50:656–658

    Article  CAS  Google Scholar 

  • McCormick SD, Sundell K, Björnsson BT, Brown CL, Hiroi J (2003) Influence of salinity on the localization of Na+/K+-ATPase, Na+/K+/2Cl− cotransporter (NKCC) and CFTR anion channel in chloride cells of the Hawaiian goby (Stenogobius hawaiiensis). J Exp Biol 206:4575–4583

    Article  PubMed  CAS  Google Scholar 

  • McCormick SD, Regish AM, Christensen AK (2009) Distinct freshwater and seawater isoforms of Na+/K+-ATPase in gill chloride cells of Atlantic salmon. J Exp Biol 212:3990–4001

    Article  Google Scholar 

  • McDowall RM (2002) The origins of the salmonid fishes: marine, freshwater…or neither? Rev Fish Biol Fisher 11:171–179

    Article  Google Scholar 

  • McDowall RM (2007) On amphidromy, a distinct form of diadromy in aquatic organisms. Fish Fisher 8:1–13

    Article  Google Scholar 

  • Mobasheri A, Avila J, Cozar-Castellano I, Brownleader MD, Trevan M, Francis MJ, Lamb JF, Martin-Vasallo P (2000) Na+, K+-ATPase isozyme diversity; comparative biochemistry and physiological implications of novel functional interactions. Biosci Rep 20:51–91

    Article  PubMed  CAS  Google Scholar 

  • Nordlie FG (2009) Environmental influences on regulation of blood plasma/serum components in teleost fishes: a review. Rev Fish Biol Fisher 19:481–564

    Article  Google Scholar 

  • Nordlie FG, Wahl WA, Binello J, Haney DC (1995) Body water content over a range of ambient salinities in the sheepshead minnow. J Fish Biol 47:624–630

    Article  Google Scholar 

  • Pagliarani A, Ventrella V, Ballestrazzi R, Trombetti F, Pirimi M, Trigari G (1991) Salinity-dependence of the properties of gill Na+, K+-ATPase in rainbow trout (Oncorhynchus mykiss). Comp Biochem Physiol 100B:229–236

    CAS  Google Scholar 

  • Rajarao SJ, Canfield VA, Mohideen MA, Yan YL, Postlethwait JH, Cheng KC, Levenson R (2001) The repertoire of Na, K-ATPase α and β subunit genes expressed in the zebrafish, Danio rerio. Genome Res 11:1211–1220

    Article  PubMed  CAS  Google Scholar 

  • Richards JG, Semple JW, Bystriansky JS, Schulte PM (2003) Na+/K+-ATPase α-isoform switching in gills of rainbow trout (Oncorhynchus mykiss) during salinity transfer. J Exp Bio 206:4475–4486

    Article  CAS  Google Scholar 

  • Schönrock C, Morley SD, Okawara Y, Lederis K, Richter D (1991) Sodium and potassium ATPase of the teleost fish Catostomus commersoni. Sequence, protein structure and evolutionary conservation of the alpha-subunit. Biol Chem Hoppe-Seyler 372:179–286

    Article  Google Scholar 

  • Scott GR, Richards JG, Forbush B, Isenring P, Schulte PM (2004) Changes in gene expression in gills of the euryhaline killifish Fundulus heteroclitus after abrupt salinity transfer. Am J Physiol-Cell Physiol 287:C300–C309

    Article  PubMed  CAS  Google Scholar 

  • Semple JW, Green HJ, Schulte PM (2002) Molecular cloning and characterization of two Na/K-ATPase isoforms in Fundulus heteroclitus. Marine Biotech 4:512–519

    Article  CAS  Google Scholar 

  • Sokal RR, Rohlf FJ (1995) Biometry: the principles and practice of statistics in biological research, 3rd edn. WH Freeman, New York

    Google Scholar 

  • Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: Molecular Evolutionary Genetics Analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739

    Article  PubMed  CAS  Google Scholar 

  • Tipsmark CK, Breves JP, Seale AP, Lerner DT, Hirano T, Grau EG (2011) Switching of Na+, K+-ATPase isoforms by salinity and prolactin in the gill of a cichlid fish. J Endocrinol 209:237–244

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors wish to thank Dr. Carol Bucking for useful discussions and technical support. We are also grateful for the feedback from two anonymous reviewers. Funding was provided by a Marsden Grant from the Royal Society of New Zealand [UOC0711], and by a Ph.D. scholarship from the Chilean Government, CONICYT, to MAU. Support for work performed at U.B.C. was provided by a travelling fellowship from The Journal of Experimental Biology and The Company of Biologists Ltd. to MAU.

Author information

Authors and Affiliations

  1. School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch, 8140, New Zealand

    Mauricio A. Urbina & Chris N. Glover

  2. Department of Zoology, The University of British Columbia, Vancouver, BC, V6T 1Z4, Canada

    Patricia M. Schulte

  3. Department of Biological Sciences, DePaul University, Chicago, IL, 60614, USA

    Jason S. Bystriansky

Authors
  1. Mauricio A. Urbina
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Patricia M. Schulte
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jason S. Bystriansky
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chris N. Glover
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mauricio A. Urbina.

Additional information

Communicated by I.D. Hume.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Urbina, M.A., Schulte, P.M., Bystriansky, J.S. et al. Differential expression of Na+, K+-ATPase α-1 isoforms during seawater acclimation in the amphidromous galaxiid fish Galaxias maculatus . J Comp Physiol B 183, 345–357 (2013). https://doi.org/10.1007/s00360-012-0719-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00360-012-0719-y

Keywords

Search

Navigation