Aquaculture International

, Volume 22, Issue 3, pp 1149–1161 | Cite as

Combined effects of acute thermal and hypo-osmotic stresses on osmolality and hsp70, hsp90 and sod expression in the sea cucumber Apostichopus japonicus Selenka

  • Qing-lin Wang
  • Shan-shan Yu
  • Chuan-xin Qin
  • Shuang-lin Dong
  • Yun-wei Dong


The combined effects of acute temperature and salinity on osmolality, expressions of heat shock proteins mRNA (hsp70, hsp90a and hsp90b) and superoxide dismutase mRNA (sod) were investigated in the sea cucumber Apostichopus japonicus Selenka. There were 12 treatments (combinations of temperature at 16, 20, 24 and 28 °C and salinity at 22, 27 and 32 ppt). In low salinity environments, the osmolality of the sea cucumber’s coelomic fluid decreased immediately and reached osmotic balance within 6 h. The decline of osmolality after 2 h of hypo-osmotic stress was faster at high temperatures (28 °C) than that at low temperatures (16 and 20 °C). Cellular level stress was indicated by up-regulation of hsp70, hsp90s and sod mRNA, and the maximal expression of all genes occurred at 6 h after stresses. The up-regulation of hsps and sod mRNA indicated the emergence of protein denaturation and oxidative damage and also suggested an increase in energy consumption at high temperature and low salinity. These results indicated that high temperature and low salinity could change biochemical pathways and energy budgets and then potentially impair the osmoregulation of the sea cucumber. Therefore, effective ways should be taken (e.g., draining off the upper freshwater, exchanging water and adding man-made sea water) to prevent the damage to sea cucumber culture caused by low salinity induced by rainstorms, especially at high temperature.


Sea cucumber Apostichopus japonicus Temperature Salinity Osmolality Heat shock proteins Superoxide dismutase 



This work was supported by grants from National Basic Research Program of China (2013CB956500), Nature Science funds for Distinguished Young Scholars of Fujian Province, China (2011J06017), National Natural Science Foundation of China (41076083, 41276126), the Fundamental Research Funds for the Central Universities, and Program for New Century Excellent Talents in University of Fujian Province, the China Postdoctoral Science Foundation (2013M541862), the open funds of Scientific Observing and Experimental Station of South China Sea Fishery Resources and Environment, Ministry of Agriculture, P. R. China (SSCS-201207). We thank Dr. Colin Little for preparing the manuscript.


  1. Balshaw DM, Millette LA, Wallick ET (2001) Sodium pump function. In: Sperelakis N (ed) Cell physiology source book: a molecular approach. Academic Press, San Diego, pp 261–268CrossRefGoogle Scholar
  2. Binyon J (1972) Salinity tolerance and ironic regulation. In: Boolootian RA (ed) Physiology of echinoderms. Pergamon Press, Oxford, pp 33–43CrossRefGoogle Scholar
  3. Castilho PC, Martins IA, Bianchini A (2001) Gill Na +/K + -ATPase and osmoregulation in the estuarine crab, Chasmagnathus granulata Dana, 1851 (Decapoda, Grapsidae). J Exp Mar 256:215–227CrossRefGoogle Scholar
  4. Charmantier G (1998) Ontogeny of osmoregulation in crustaceans: a review. Invertebr Reprod Dev 33:177–190CrossRefGoogle Scholar
  5. Diehl WJ (1986) Osmoregulation in echinoderms. Comp Biochem Physiol 84A:199–205CrossRefGoogle Scholar
  6. DOF (Department of Fisheries) (2012) China fisheries statistic yearbook. China Agriculture Press, BeijingGoogle Scholar
  7. Dong YW, Dong SL (2008) Induced thermotolerance and expression of heat shock protein 70 in sea cucumber Apostichopus japonicus. Fish Sci 74:573–578CrossRefGoogle Scholar
  8. Dong YW, Dong SL, Ji TT (2008a) Effect of different thermal regimes on growth and physiological performance of the sea cucumber Apostichopus japonicus Selenka. Aquaculture 275:329–334CrossRefGoogle Scholar
  9. Dong YW, Dong SL, Meng XL (2008b) Effects of thermal and osmotic stress on growth, osmoregulation and Hsp70 in sea cucumber (Apostichopus japonicus Selenka). Aquaculture 276:179–186CrossRefGoogle Scholar
  10. Dong YW, Ji TT, Meng XL et al (2010) Difference in thermotolerance between green and red color variants of the Japanese sea cucumber, Apostichopus japonicus Selenka: Hsp70 and heat-hardening effect. Biol Bull 218:87–94PubMedGoogle Scholar
  11. Dutton JM, Hofmann GE (2009) Biogeographic variation in Mytilus galloprovincialis heat shock gene expression across the eastern Pacific range. J Exp Mar Biol Ecol 376:37–42CrossRefGoogle Scholar
  12. Feder ME, Hofmann GE (1999) Heat shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. Ann Rev Physiol 61:243–282CrossRefGoogle Scholar
  13. Feige V, Morimoto RI, Yahara I et al (1996) Stress inducible cellular responses. Birkhauser, BostonGoogle Scholar
  14. Frydman J, Höhfeld J (1997) Chaperones get in touch: the hip-hop connection. Trends Biochem Sci 22:87–92PubMedCrossRefGoogle Scholar
  15. Fu XY, Xue CH, Miao BC et al (2005) Characterization of proteases from the digestive tract of sea cucumber (Stichopus japonicas): high alkaline protease activity. Aquaculture 246:321–329CrossRefGoogle Scholar
  16. Hartl FU (1996) Molecular chaperones in protein folding. Nature 381:571–580PubMedCrossRefGoogle Scholar
  17. Imsland AK, Gunnarsson S, Foss A et al (2003) Gill Na +/K + -ATPase activity, plasma chloride and osmolality in juvenile turbot (Scophthalmus maximus) reared at different temperatures and salinities. Aquaculture 218:671–683CrossRefGoogle Scholar
  18. Ji TT, Dong YW, Dong SL (2008) Growth and physiological responses in the sea cucumber, Apostichopus japonicus Selenka: aestivation and temperature. Aquaculture 283:180–187CrossRefGoogle Scholar
  19. Jorgensen PL, Håkansson KO, Karlish SJD (2003) Structure and mechanism of Na +/K + -ATPase: functional sites and their interactions. Ann Rev Physiol 65:817–849CrossRefGoogle Scholar
  20. Kinne O (1971) Salinity: 3. Animals: 1. Invertebrates. In: Kinne O (ed) Marine ecology: a comprehensive, integrated treatise on life in oceans and coastal waters: 1. Environmental factors, 2, pp 821–995Google Scholar
  21. Leiniö S, Lehtonen KK (2005) Seasonal variability in biomarkers in the bivalves Mytilus edulis and Macoma balthica from the northern Baltic Sea. Comp Biochem Physiol 140C:408–421Google Scholar
  22. Lemaire P, Bernard E, Martinez-Paz J et al (2002) Combined effect of temperature and salinity on osmoregulation of juvenile and subadult Penaeus stylirostris. Aquaculture 209:307–317CrossRefGoogle Scholar
  23. Meng XL, Ji TT, Dong YW et al (2009) Thermal resistance in sea cucumbers (Apostichopus japonicus) with differing thermal history: the role of Hsp70. Aquaculture 294:314–318CrossRefGoogle Scholar
  24. Meng XL, Dong YW, Dong SL et al (2011) Mortality of the sea cucumber, Apostichopus japonicus Selenka, exposed to acute salinity decrease and related physiological responses: osmoregulation and heat shock protein expression. Aquaculture 316:88–92CrossRefGoogle Scholar
  25. Morimoto RI (1998) Regulation of the heat shock transcriptional response: cross talk between a family of heat shock factors, molecular chaperones, and negative regulators. Gene Dev 12:3788–3796PubMedCrossRefGoogle Scholar
  26. Morimoto RI, Tissières A, Georgopoulos C (1990) Stress proteins in biology and medicine. Cold Spring Harbor, NYGoogle Scholar
  27. Niu C, Rummer J, Brauner C et al (2008) Heat shock protein (Hsp70) induced by a mild heat shock slightly moderates plasma osmolarity increases upon salinity transfer in rainbow trout (Oncorhynchus mykiss). Comp Biochem Physiol 148C:437–444Google Scholar
  28. Pan F, Zarate J, Tremblay G et al (2000) Cloning and characterization of salmon hsp90 cDNA: upregulation by thermal and hyperosmotic stress. J Exp Zool 287:199–212PubMedCrossRefGoogle Scholar
  29. Parihar MS, Javeri T, Hemnani T et al (1997) Response of superoxide dismutase, glutathione antioxidant defenses in gills of the freshwater catfish (Heteropneustes Fossilis) to short-term elevated temperature. J Therm Biol 22(2):151–156CrossRefGoogle Scholar
  30. Ruppert EE, Barnes RD, Fox RS (1994) Invertebrate zoology. Saunders, Fort WorthGoogle Scholar
  31. Somero GN (2002) Thermal physiology and vertical zonation of intertidal animals: optima, limits, and cost of living. Integr Comp Biol 42:780–789PubMedCrossRefGoogle Scholar
  32. Storti RV, Scott MP, Rich A et al (1980) Translational control of protein synthesis in response to heat shock in D. melanogaster cells. Cell 22:825–834PubMedCrossRefGoogle Scholar
  33. Tadashi K, Takao K, Hiroshi A et al (2004) Mild heat shock induces autophagic growth arrest, but not apoptosis in U251-MG and U87-MG human malignant glioma cells. J Neuro Oncol 68(2):101–111CrossRefGoogle Scholar
  34. Tomanek L, Somero GN (1999) Evolutionary and acclimation-induced variation in the heat-shock responses of congeneric marine snails (genus Tegula) from different thermal habitats: implications for limits of thermotolerance and biogeography. J Exp Biol 202:2925–2936PubMedGoogle Scholar
  35. Vidolin D, Santos-Gouvea IA, Freire CA (2002) Osmotic stability of the coelomic fluids of a sea-cucumber (Holothuria grisea) and starfish (Asterina stellifera) (Echinodermata) exposed to the air during low tide. Acta Biol Par 31:113–121Google Scholar
  36. Wang QL, Dong YW, Dong SL et al (2011) Effects of heat-shock selection during pelagic stages on thermal sensitivity of juvenile sea cucumber Apostichopus japonicus Selenka. Aquac Int 19:1165–1175CrossRefGoogle Scholar
  37. Wang QL, Dong YW, Qin CX et al (2012) Effects of rearing temperature on growth, metabolism and thermal tolerance of juvenile sea cucumber, Apostichopus japonicus Selenka: critical thermal maximum (CTmax) and hsps gene expression. Aquac Res (press). doi: 10.1111/j.1365-2109.2012.03162.x)
  38. Wilhelm Filho D, Tribess T, Gáspari C et al (2001) Seasonal changes in antioxidant defenses of the digestive gland of the brown mussel (Perna perna). Aquaculture 203:149–158CrossRefGoogle Scholar
  39. Wilhelm-Filho DW, Giulivi C, Boveris A (1993) Antioxidant defenses in marine fish. I: Teleosts. Comp Biochem Physiol 106C:409–413Google Scholar
  40. Williams AB (1960) The influence of temperature on osmotic regulation in two species of estuarine shrimps (Penaeus). Biol Bull 119:560–571CrossRefGoogle Scholar
  41. Wu ZZ (2007) Improved method of NH4-N determined by hypobromite oxidation in water. Mar Environ Sci 26(1):84–87 (in Chinese with English abstract)Google Scholar
  42. Yuan XT, Yang HS, Zhou Y et al (2006) Salinity effect on respiration and excretion of sea cucumber Apostichopus japonicus (Selenka). Oceanologia Et Limnologia Sin 37:348–354 (in Chinese with English abstract)Google Scholar
  43. Yusuke Y, Tatsuo H, Koichi M (2006) Distribution of the Japanese sea cucumber Apostichopus japonicus in the intertidal zone of Hirao Bay, eastern Yamaguchi Perf., Japan-Suitable environmental factors for juvenile habitats. J Natl Fish Univ 54:111–120Google Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Qing-lin Wang
    • 1
    • 3
  • Shan-shan Yu
    • 1
  • Chuan-xin Qin
    • 2
  • Shuang-lin Dong
    • 3
  • Yun-wei Dong
    • 1
  1. 1.State Key Laboratory of Marine Environmental Science, College of Marine and Earth SciencesXiamen UniversityXiamenPeople’s Republic of China
  2. 2.South China Sea Fisheries Research InstituteChinese Academy of Fishery ScienceGuangzhouPeople’s Republic of China
  3. 3.The Key Laboratory of Mariculture, Ministry of Education, Fisheries CollegeOcean University of ChinaQingdaoPeople’s Republic of China

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