Marine Biology

, Volume 156, Issue 9, pp 1751–1764 | Cite as

Branchial and intestinal osmoregulatory acclimation in the four-eyed sleeper, Bostrychus sinensis (Lacepède), exposed to seawater

  • W. Y. X. Peh
  • S. F. Chew
  • J. M. Wilson
  • Y. K. IpEmail author
Original Paper


Bostrychus sinensis is a facultative air breather that inhabits waters of a wide range of salinities. This study aimed to elucidate whether branchial and intestinal osmoregulatory acclimation occurred in B. sinensis transferred from 5‰ water through a progressive increase in salinities to seawater. Our results indicate that B. sinensis acted as a hyperosmotic regulator in 5‰ water, but exhibited hypoosmotic hypoionic regulation in seawater. During short- (1 day) and medium- (10 days) term acclimation to seawater, there were only minor perturbations in plasma osmolality and [Na+], which returned to control levels after 45 days of exposure to seawater. Branchial Na+/K+-ATPase activity was unaffected by 1, 10 or 45 days of exposure to seawater. However, prolonged (45 days) acclimation to seawater led to a significant increase in Na+/K+-ATPase α-subunit protein abundance. Taken together, these results indicate that there could be changes in the expression of Na+/K+-ATPase isoforms and/or post-translational modification of Na+/K+-ATPase in the gills of fish exposed to seawater. Immunofluorescence microscopy revealed that acclimation to seawater for 10 days only resulted in no change in branchial Na+/K+-ATPase protein expression, but there were increases in protein expression of cystic fibrosis transmembrane regulator (CFTR)-like chloride channel and Na+:K+:2Cl cotransporter (NKCC; probably NKCC1). Indeed, NKCC was undetectable in gills of fish kept in 5‰ water by Western blotting, but it became weakly detectable in fish exposed to seawater for 10 days and prominently expressed in fish exposed to seawater for 45 days. Therefore, our results indicate that branchial CFTR-like chloride channel and NKCC1 were the determining factors in the transition between hyperosmotic regulation and hypoosmotic hypoionic regulation in B. sinensis. Furthermore, the intestine of B. sinensis also served as an important osmoregulatory organ, since there were significant increases in both the activity and protein abundance of intestinal Na+/K+-ATPase in fish acclimated to seawater for 45 days. The effectiveness of branchial and intestinal osmoregulatory acclimation in B. sinensis during seawater acclimation led to only a minor increase in plasma osmolality, and thus resulted in relatively unchanged free amino acid contents in muscle and liver.


ATPase Activity Plasma Osmolality Cystic Fibrosis Transmembrane Regulator TFAA Euryhaline Fish 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This project is supported by the Ministry of Education of the Republic of Singapore through grants R-154-000-409-112.


  1. Anderson PM, Broderius MA, Fong KC, Tsui KNT, Chew SF, Ip YK (2002) Glutamine synthetase expression in liver, muscle, stomach and intestine of Bostrichyths sinensis in response to exposure to a high exogenous ammonia concentration. J Exp Biol 205:2053–2065PubMedGoogle Scholar
  2. Bergmeyer HU, Beutler HO (1985) Ammonia. In: Bergmeyer HU, Bergmeyer J, Graβl M (eds) Methods of enzymatic analysis, vol VIII. Verlag Chemie, Weinheim, pp 454–461Google Scholar
  3. Bradford MM (1976) A rapid and sensitive method of the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  4. Bystrainsky JS, Richards JG, Schulte PM, Ballantyne JS (2006) Reciprocal expression of gill Na+/K+-ATPase α-subunit isoforms 1a and 1b during seawater acclimation of three salmonid fishes that vary in their salinity tolerance. J Exp Biol 209:1848–1858CrossRefGoogle Scholar
  5. Chang EWY, Loong AM, Wong WP, Chew SF, Wilson JM, Ip YK (2007) Changes in tissue free amino acid contents, branchial Na+/K+-ATPase activity and bimodal breathing pattern in the freshwater climbing perch, Anabas testudineus (Bloch), during seawater acclimation. J Exp Zool 307A:708–723CrossRefGoogle Scholar
  6. Cutler CP, Cramb G (2002) Two isoforms of the Na+/K+/2Cl cotransporter are expressed in the European eel (Anguilla anguilla). Biochim Biophys Acta Biomembrane 1566:92–103CrossRefGoogle Scholar
  7. Cutler CP, Sanders IL, Hazon N, Cramb G (1995a) Primary sequence, tissue specificity and expression of the Na+, K+-ATPase α1 subunit in the European eel (Anguilla anguilla). Comp Biochem Physiol 111B:567–573CrossRefGoogle Scholar
  8. Cutler CP, Sanders IL, Hazon N, Cramb G (1995b) Primary sequence, tissue-specificity and messenger-RNA expression of the Na+, K+-ATPase β1 subunit in the European eel (Anguilla anguilla). Fish Physiol Biochem 14:423–429CrossRefGoogle Scholar
  9. 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 85:97–177CrossRefGoogle Scholar
  10. Fiess JC, Kunkel-Patterson A, Mathias L, Riley LG, Yancey PH, Hirano T, Grau EG (2007) Effects of environmental salinity and temperature on osmoregulatory ability, organic osmolytes, and plasma hormone profiles in the Mozambique tilapia (Oreochromis mossambicus). Comp Biochem Physiol A 146:252–264CrossRefGoogle Scholar
  11. Fuentes J, Soengas JL, Rey P, Rebolledo E (1997) Progressive transfer to seawater enhances intestinal and branchial Na+–K+ ATPase activity in nonanadromous rainbow trout. Aquacult Int 5:217–227CrossRefGoogle Scholar
  12. Gamba G (2005) Molecular physiology and pathophysiology of electroneutral cation-chloride cotransporters. Physiol Rev 85:423–493CrossRefGoogle Scholar
  13. Goss GG, Adamia S, Galvez F (2001) Peanut lectin binds to a subpopulation of mitochondria-rich cells in the rainbow trout gill epithelium. Am J Physiol Regulatory Integrative Comp Physiol 281:R1718–R1725CrossRefGoogle Scholar
  14. Graham JB (1997) Air-breathing fishes: evolution, diversity, and adaptation. Academic Press, San DiegoCrossRefGoogle Scholar
  15. Grosell M (2006) Intestinal anion exchange in marine fish osmoregulation. J Exp Biol 209:2813–2827CrossRefGoogle Scholar
  16. Grosell M, Wood CM, Wilson RW, Bury NR, Hogstrand C, Rankin JC, Jensen FB (2005) Bicarbonate secretion plays a role in chloride and water absorption of the European flounder intestine. Am J Physiol 288:R936–R946Google Scholar
  17. Hiroi J, Yasumasu S, McCormick SD, Hwang PP, Kaneko T (2008) Evidence for an apical Na-Cl cotransporter involved in ion uptake in a teleost fish. J Exp Biol 211:2584–2599CrossRefGoogle Scholar
  18. Huang Z (2001) Marine species and their distribution in China’s seas. In: Vertebrata. Smithsonian Institution, Florida, pp 404–463Google Scholar
  19. Huggins AK, Colley L (1971) The changes in the nonprotein nitrogenous constituents of muscle during the adaptation of the eel Anguilla anguilla L. from fresh water to sea water. Comp Biochem Physiol 38B:537–541Google Scholar
  20. Hwang PP, Lee TH (2007) New insights into fish ion regulation and mitochondrion-rich cells. Comp Biochem Physiol A 148:479–497CrossRefGoogle Scholar
  21. Hwang HC, Chen IY, Yueh PC (1988) The freshwater fishes of China in colored illustrations, vol 2. Shanghai Sciences and Technology Press, Shanghai, p 201Google Scholar
  22. Ip YK, Chew SF, Leong IWA, Jin Y, Wu RSS, Lim CB (2001) The sleeper Bostrichyths sinensis (Teleost) stores glutamine and reduces ammonia production during aerial exposure. J Comp Physiol B 171:357–367CrossRefGoogle Scholar
  23. Jow LY, Chew SF, Lim CB, Anderson PM, Ip YK (1999) The marble goby, Oxyeleotris marmoratus, activates hepatic glutamine synthetase and detoxifies ammonia to glutamine during terrestrial exposure. J Exp Biol 202:237–245PubMedGoogle Scholar
  24. Kaushik SJ, Luquet P (1979) Influence of dietary amino acid patterns on the free amino acid contents of blood and muscle of rainbow trout (Salmo gairdnerri R). Comp Biochem Physiol 64B:175–180Google Scholar
  25. Kottelat M, Whitten AJ, Kartikasari SN, Wirjoatmodjo S (1993) Freshwater fishes of Western Indonesia and Sulawesi. Periplus Editions, Hong Kong, p 221Google Scholar
  26. Kuo SR, Shao KT (1999) Species composition of fish in the coastal zones of the Tsengwen estuary, with descriptions of five new records from Taiwan. Zool Stud 38:391–404Google Scholar
  27. Lin CH, Lee TH (2005) Sodium or potassium ions activate different kinetics of gill Na, K-ATPase in three seawater and freshwater-acclimated euryhaline teleosts. J Exp Zool 303A:57–65CrossRefGoogle Scholar
  28. 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 A 138:287–295CrossRefGoogle Scholar
  29. Lytle C, Xu JC, Biemesderfer D, Forbush B III (1995) Distribution and diversity of Na-K-Cl cotransport proteins: a study with monoclonal antibodies. Am J Physiol 269:C1496–C1505CrossRefGoogle Scholar
  30. Mackay WC, Janicki R (1978) Changes in the eel intestine during seawater adaptation. Comp Biochem Physiol 62A:757–761Google Scholar
  31. Marshall WS, Grosell M (2006) Ion transport, osmoregulation and acid-base balance. In: The physiology of fishes, 3rd edn, pp 177–230Google Scholar
  32. McCormick SD, Saunders RL (1987) Preparatory physiological adaptations for marine life of salmonids: osmoregulation, growth and metabolism. Am Fish Soc Symp 94A:95–97Google Scholar
  33. McCormick SD, Sundell K, Björnsson BT, Brown CL, Hiroi J (2003) Influence of salinity on the localization of Na+/K+-ATPase, Na+/K+/2Clcotransporter (NKCC) and CFTR anion channel in chloride cells of the Hawaiian goby (Stenogobius hawaiiensis). J Exp Biol 206:4575–4583CrossRefGoogle Scholar
  34. Nguyen NT, Nguyen VQ (2006) Biodiversity and living resources of the coral reef fishes in Vietnam marine waters. Science and Technology Publishing House, HanoiGoogle Scholar
  35. Ni IH, Kwok KY (1999) Marine fish fauna in Hong Kong waters. Zool Stud 38:130–152Google Scholar
  36. Potts WTW, Evans DH (1967) The effects of hypophysectomy and bovine prolactin on salt fluxes in fresh-water-adapted Fundulus heteroclitus. Biol Bull 131:362–368CrossRefGoogle Scholar
  37. 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 Biol 206:4475–4486CrossRefGoogle Scholar
  38. Scott GR, Claiborne JB, Edwards SL, Schulte PM, Wood CM (2005) Gene expression after freshwater transfer in gills and opercular epithelia of killifish: insight into divergent mechanisms of ion transport. J Exp Biol 208:2719–2729CrossRefGoogle Scholar
  39. Seidelin M, Madsen SS, Blenstrup H, Tipsmark CK (2000) Time-course changes in the expression of the Na+, K+-ATPase in gills and pyloric caeca of brown trout (Salmo trutta) during acclimation to seawater. Physiol Biochem Zool 73:446–453CrossRefGoogle Scholar
  40. Semple JW, Green HJ, Schulte PM (2002) Molecular cloning and characterization of two Na/K-ATPase isoforms in Fundulus heteroclitus. Marine Biotech 4:512–519CrossRefGoogle Scholar
  41. Tipsmark CK, Madsen SS, Seidelin M, Christensen AS, Cutler CP, Cramb G (2002) Dynamics of Na+, K+, 2Cl cotransporter and Na+, K+-ATPase expression in the branchial epithelium of brown trout (Salmo trutta) and Atlantic salmon (Salmo salar). J Exp Zool 293:106–118CrossRefGoogle Scholar
  42. Tok CY, Chew SF, Peh WYX, Loong AM, Wong WP, Ip YK (2009) Glutamine accumulation and up-regulation of glutamine synthetase activity in the swamp eel, Monopterus albus (Zuiew), exposed to brackish water. J Exp Biol 212:1248–1258CrossRefGoogle Scholar
  43. Tse WKF, Au DWT, Wong CKC (2006) Characterization of ion channel and transporter mRNA expressions in isolated gill chloride and pavement cells of seawater acclimating eels. Biochem Biophys Res Commun 346:1181–1190CrossRefGoogle Scholar
  44. Usher ML, Talbot C, Eddy FB (1991) Intestinal water transport in juvenile atlantic salmon (Salmo salar L.) during smolting and following transfer to seawater. Comp Biochem Physiol 100A:813–818CrossRefGoogle Scholar
  45. Wilson RW, Grosell M (2003) Intestinal bicarbonate secretion in marine teleost fish—source of bicarbonate, pH sensitivity, and consequence for whole animal acid-base and divalent cation homeostasis. Biochim Biophys Acta 1618:163–193CrossRefGoogle Scholar
  46. Wilson RW, Gilmour K, Henry R, Wood CM (1996) Intestinal base excretion in the seawater-adapted rainbow trout: a role in acid–base balance? J Exp Biol 199:2331–2343PubMedGoogle Scholar
  47. Wilson RW, Wilson JM, Grosell M (2002) Intestinal bicarbonate secretion by marine teleost fish—why and how? Biochim Biophys Acta 1566:182–193CrossRefGoogle Scholar
  48. Wilson JM, Leitão A, Gonçalves AF, Ferreira C, Reis-Santos P, Fonseca A-V, da Silva JM, Antunes JC, Pereira-Wilson C, Coimbra J (2007) Modulation of branchial ion transport protein expression by salinity in glass eels (Anguilla anguilla L.). Mar Biol 151:1633–1645CrossRefGoogle Scholar
  49. Wu YC, Lin LY, Lee TH (2003) Na+/K+/2Cl cotransporter: a novel marker for identifying freshwater- and seawater-type mitochondria-rich cells in gills of the euryhaline tilapia, Oreochromis mossambicus. Zool Stud 42:186–192Google Scholar
  50. Yancey PH (2001) Nitrogen compounds as osmolytes. In: Wright PA, Anderson PM (eds) Fish physiology: nitrogen excretion, vol 20. Academic Press, New York, pp 309–341CrossRefGoogle Scholar
  51. Zaugg WS (1982) A simplified preparation for adenosine triphosphatase determination in gill tissue. Can J Fish Aquat Sci 39:215–217CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • W. Y. X. Peh
    • 1
  • S. F. Chew
    • 2
  • J. M. Wilson
    • 3
  • Y. K. Ip
    • 1
    Email author
  1. 1.Department of Biological ScienceNational University of SingaporeSingaporeRepublic of Singapore
  2. 2.Natural Sciences and Science Education, National Institute of EducationNanyang Technological UniversitySingaporeRepublic of Singapore
  3. 3.Ecofisiologia CIMARPortoPortugal

Personalised recommendations