Fish Physiology and Biochemistry

, Volume 43, Issue 1, pp 89–102 | Cite as

hsp90 and hsp47 appear to play an important role in minnow Puntius sophore for surviving in the hot spring run-off aquatic ecosystem

  • Arabinda Mahanty
  • Gopal Krishna Purohit
  • Ravi Prakash Yadav
  • Sasmita Mohanty
  • Bimal Prasanna MohantyEmail author


Changes in the expression of a number of hsp genes in minnow Puntius sophore collected from a hot spring run-off (Atri hot spring in Odisha, India; 20o09′N 85°18′E, 36–38 °C) were investigated to study the upper thermal acclimation response under heat stress, using same species from aquaculture ponds (water temperature 27 °C) as control. Expression of hsp genes was analyzed in both groups using RT-qPCR, which showed up-regulation of hsp90 (2.1-fold) and hsp47 (2.5-fold) in hot spring run-off fishes, whereas there was no alteration in expression of other hsps. As the fish inhabit the hot spring run-off area for very long duration, they could have adapted to the environment. To test this hypothesis, fishes collected from hot spring run-off were divided into two groups; one was heat-shocked at 41 °C/24 h, and the other was acclimatized at 27 °C/24 h. Up-regulation of all the hsps (except hsp78) was observed in the heat-shocked fishes, whereas expression of all hsps was found to be down-regulated to the basal level in fishes maintained at 27 °C/24 h. Pathway analysis showed that the expressions of all the hsps except hsp90 are regulated by the transcription factor heat shock factor 1 (Hsf1). This study showed that hsp90 and hsp47 play an important role in Puntius sophore for surviving in the high-temperature environment of the hot spring run-off. Additionally, we show that plasticity in hsp gene expression is not lost in the hot spring run-off population.


Heat stress Thermal acclimation hsp gene Gene plasticity RT-qPCR 



This research was funded by the Indian Council of Agricultural Research under the National Fund for Basic, Strategic and Frontier Application Research in Agriculture (NFBSFARA; recently renamed National Agricultural Science Fund, NASF) Project # AS-2001 (B.P.M. and S.M). A.M. and G.K.P. are NFBSFARA Senior Research Fellows. The authors are thankful to Director, ICAR—Central Inland Fisheries Research Institute, Barrackpore, and Director, School of Biotechnology, KIIT University, Bhubaneswar, for the facilities and encouragement. Technical assistance received from Shri Laddu Ram Mahaver, Samir K. Paul and Rabiul Sk. is acknowledged. The authors would like to acknowledge the anonymous reviewers for critically reviewing the manuscript; the constructive criticism and suggestions from the reviewers have resulted in substantial improvement of the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interests.

Supplementary material

10695_2016_270_MOESM1_ESM.doc (58 kb)
Supplementary material 1 (DOC 58 kb)
10695_2016_270_MOESM2_ESM.doc (47 kb)
Supplementary material 2 (DOC 47 kb)


  1. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215(3):403–410CrossRefPubMedGoogle Scholar
  2. Arrigo A (2007) The cellular “networking” of mammalian HSP27 and its functions in the control of protein folding, redox state and apoptosis. Adv Exp Med Biol 594:14–26CrossRefPubMedGoogle Scholar
  3. Barnes JA, Dix DJ, Collins BW, Luft C, Allen JW (2001) Expression of inducible Hsp70 enhances the proliferation of MCF-7 breast cancer cells and protects against the cytotoxic effects of hyperthermia. Cell Stress Chaperone 6(4):316–325CrossRefGoogle Scholar
  4. Basu N, Nakano T, Grau EG, Iwama GK (2001) The effects of cortisol on heat shock protein 70 levels in two fish species. Gen Comp Endocrinol 124:97–105CrossRefPubMedGoogle Scholar
  5. Bonham RT, Fine MR, Pollock FM, Shelden EA (2003) HSP27, HSP70, and metallothionein in MDCK and LLC-PK1 renal epithelial cells: effects of prolonged exposure to cadmium. Toxicol Appl Pharmacol 191(1):63–73CrossRefPubMedGoogle Scholar
  6. Boyd CE (1998) Water quality for pond aquaculture. International Center for Aquaculture and Aquatic Environment, Auburn University, Auburn, Albania
  7. Brown SA, Kingston RE (1997) Disruption of downstream chromatin directed by a transcriptional activator. Genes Dev 11:3116–3121CrossRefPubMedPubMedCentralGoogle Scholar
  8. Buckley BA, Hofmann GE (2002) Thermal acclimation changes DNA-binding activity of heat shock factor 1(HSF1) in the goby Gillichthys mirabilis: implications for plasticity in the heat shock response in natural populations. J Exp Biol 205:3231–3240PubMedGoogle Scholar
  9. Colson-Proch C, Morales A, Hervant F, Konecny L, Moulin C, Douady CJ (2010) First cellular approach of the effects of global warming on groundwater organisms: a study of the HSP70 gene expression. Cell Stress Chaperones 15(3):259–270CrossRefPubMedGoogle Scholar
  10. CPCB (2007) Guidelines for water quality monitoring. Central Pollution Control Board, New DelhiGoogle Scholar
  11. Cuesta R, Laroia G, Schneider RJ (2000) Chaperone HSP27 inhibits translation during heat shock by binding eIF4G and facilitating dissociation of cap-initiation complexes. Genes Dev 14(12):1460–1470PubMedPubMedCentralGoogle Scholar
  12. Dahanukar N (2010) Puntius sophore. The IUCN Red List of Threatened Species 2010: e.T166623A6249514.
  13. Dale Becker C, Genoway RG (1979) Evaluation of the critical thermal maximum for determining thermal tolerance of freshwater fish. Environ Biol Fish 4(3):245–256CrossRefGoogle Scholar
  14. Das T, Pal AK, Chakraborty SK, Manusha SM, Mukherjee SC (2005) Thermal tolerance and oxygen consumption of Indian major carps acclimated to four temperatures. J Therm Biol 29(3):157–163CrossRefGoogle Scholar
  15. Das A, Panda SS, Palita SK, Patra HK, Dhal NK (2012) Spatial and temporal variation of phytoplanktons in hot spring of Atri, Odisha, India. Curr Bot 3(5):35–40Google Scholar
  16. DeBolt S (2010) Copy number variation shapes genome diversity in Arabidopsis over immediate family generational scales. Genome Biol Evol 2:441–453CrossRefPubMedPubMedCentralGoogle Scholar
  17. Denlinger DL, Rinehart JP, Yocum GD (2001) Stress proteins: a role in insect diapause? In: Denlinger DL, Giebultowicz J, Saunders DS (eds) Insect timing: circadian rhythmicity to seasonality. Elsevier, Amsterdam, pp 155–171CrossRefGoogle Scholar
  18. Dong Y, Dong S, Ji T (2008) Effect of different thermal regimes on growth and physiological performance of the sea cucumber Apostichopus japonicus Selenka. Aquaculture 275(1–4):329–334CrossRefGoogle Scholar
  19. Elliot JM, Elliot JA (1995) The effect of the rate of temperature increase on the critical thermal maximum for parr of Atlantic salmon and brown trout. J Fish Biol. doi: 10.1111/j.1095-8649.1995.tb06014.x Google Scholar
  20. Fangue NA, Hofmeister M, Schulte PM (2006) Intraspecific variation in thermal tolerance and heat shock protein gene expression in common killifish, Fundulus heteroclitus. J Exp Biol 209:2859–2872CrossRefPubMedGoogle Scholar
  21. 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
  22. Fink AL (1999) Chaperone-mediated protein folding. Physiol Rev 79(2):425–449PubMedGoogle Scholar
  23. Haslbeck M, Vierling E (2015) A first line of stress defense: small heat shock proteins and their function in protein homeostasis. J Mol Bio 427(7):1537–1548CrossRefGoogle Scholar
  24. Iwama GK, Vijayan MM, Forsyth RB, Ackerman PA (1999) Heat shock proteins and physiological stress in fish. Am Zool 39:901–909CrossRefGoogle Scholar
  25. Iwama GK, Afonso LOB, Todgham A, Ackerman P, Kazumi Nakano K (2004) Are hsps suitable for indicating stressed states in fish? J Exp Biol 207:15–19CrossRefPubMedGoogle Scholar
  26. James TC, Usher J, Campbell S, Bond U (2008) Lager yeasts possess dynamic genomes that undergo rearrangements and gene amplification in response to stress. Curr Genet 53:139–152CrossRefPubMedGoogle Scholar
  27. Kikuchi K, Yamashita M, Watabe S, Aida K (1995) The warm temperature acclimation-related 65-kDa protein, Wap65, in goldfish and its gene expression. J Biol Chem 270(29):17087–17092CrossRefPubMedGoogle Scholar
  28. Kondrashov FA (2012) Gene duplication as a mechanism of genomic adaptation to a changing environment. Proc R Soc B. doi: 10.1098/rspb.2012.1108 Google Scholar
  29. Kregel KCJ (2002) Heat shock proteins: modifying factors in physiological stress responses and acquired thermotolerance. J Appl Physiol 92(5):2177–2186CrossRefPubMedGoogle Scholar
  30. Kubista M, Andrade JM, Bengtsson M, Forootan A, Jonak J, Lind K et al (2006) The real-time polymerase chain reaction. Mol Asp Med 27:95e125CrossRefGoogle Scholar
  31. Leach MD, Budge S, Walker L, Munro C, Cowen LE, Brown AJP (2012) Hsp90 orchestrates transcriptional regulation by Hsf1 and cell wall remodelling by MAPK signalling during thermal adaptation in a pathogenic yeast. PLoS ONE 8(12):e1003069Google Scholar
  32. Li J, Soroka J, Buchner J (2012) The Hsp90 chaperone machinery: conformational dynamics and regulation by co-chaperones. Biochim Biophys Acta 1823(3):624–635CrossRefPubMedGoogle Scholar
  33. Lindquist S, Craig E (1988) The heat-shock proteins. Annu Rev Genet 22:631–677CrossRefPubMedGoogle Scholar
  34. Logan CA, Buckley BA (2015) Transcriptomic responses to environmental temperature in eurythermal and stenothermal fishes. J Exp Biol 218:1915–1924CrossRefPubMedGoogle Scholar
  35. Mahanty A, Ganguly S, Verma A, Sahoo S, Paria P, Mitra P, Singh BK, Sharma AP, Mohanty BP (2014) Nutrient profile of small indigenous fish Puntius sophore: proximate composition, amino acid, fatty acid and micronutrient profiles. Natl Acad Sci Lett 37(1):39–44CrossRefGoogle Scholar
  36. Mailhos C, Howard MK, Latchman DS (1994) Heat shock proteins hsp90 and hsp70 protect neuronal cells from thermal stress but not from programmed cell death. J Neurochem 63(5):1787–1795CrossRefPubMedGoogle Scholar
  37. Martínez-Paz P, Morales M, Martín R, Martínez-Guitarte JL, Morcillo G (2014) Characterization of the small heat shock protein Hsp27 gene in Chironomus riparius (Diptera) and its expression profile in response to temperature changes and xenobiotic exposures. Cell Stress Chaperones 19:529–540CrossRefPubMedGoogle Scholar
  38. McDiarmid RW, Altig R (1999) Tadpoles: the biology of anuran larvae. University of Chicago Press, Chicago, p 202Google Scholar
  39. Meffe GK, Weeks SC, Mulvey M, Kandl KL (1995) Genetic differences in thermal tolerance of eastern mosqyitofish (Gambusia holbrooki; Poeciliidae) from ambient and thermal ponds. Can J Fish Aquat Sci 52(12):2704–2727CrossRefGoogle Scholar
  40. Meyer A, Biermann CH, Orti G (1993) The phylogenetic position of the zebrafish (Danio rerio), a model system in developmental biology: an invitation to the comparative method. Proc Biol Sci 252(1335):231–236CrossRefPubMedGoogle Scholar
  41. Mohanty S, Mahanty A, Yadav RP, Purohit GK, Mohanty BN, Mohanty BP (2014) The Atri hot spring in Odisha—a natural ecosystem for global warming research. Int J Geol Earth Environ Sci 4:85–90Google Scholar
  42. Mohanty BP, Mitra T, Banerjee S, Bhattacharjee S, Mahanty A, Ganguly S, Purohit GK, Karunakaran D, Mohanty S (2015) Proteomic profiling of white muscle from freshwater catfish Rita rita. Physiol Biochem 41:789–802Google Scholar
  43. Mounier N, Arrigo AP (2002) Actin cytoskeleton and small heat shock proteins: how do they interact? Cell Stress Chaperone 7(2):167–176CrossRefGoogle Scholar
  44. Nakano K, Iwama GK (2002) The 70- kDa heat shock protein response in two intertidal sculpins, Oligocottus maculosus and O. snyderi: relationship of hsp70 and thermal tolerance. Comp Biochem Physiol A 133:79–94CrossRefGoogle Scholar
  45. Norris CE, diLorio PJ, Schultz RJ, Hightower LE (1995) Variation in heat shock protein within tropical and desert species of Poeciliid fishes. Mol Biol Evol 12(6):1048–1062PubMedGoogle Scholar
  46. Ojima N, Yamashita M, Watabe S (2005) Quantitative mRNA expression profiling of heat-shock protein families in rainbow trout cells. Biochem Biophys Res Commun 329(1):51–57CrossRefPubMedGoogle Scholar
  47. Oksala NKJ, Ekmekçi FG, Ozsoy E, Kirankaya S, Kokkola T, Emecen G, Lappalainen J, Kaarniranta K, Atalay M (2014) Natural thermal adaptation increases heat shock protein levels and decreases oxidative stress. Redox Biol 3:25–28CrossRefPubMedPubMedCentralGoogle Scholar
  48. Panda SK, Jyoti V, Bhadra B, Nayak KC, Shivaji S, Rainey FA, Das SK (2009) Thiomonas bhubaneswarensis sp. nov., an obligately mixotrophic, moderately thermophilic, thiosulfate-oxidizing bacterium. Int J Syst Evol Microbiol 59:2171–2175CrossRefPubMedGoogle Scholar
  49. Pearl LH, Prodromou C (2006) Structure and mechanism of the Hsp90 molecular chaperone machinery. Annu Rev Biochem 75:271–294CrossRefPubMedGoogle Scholar
  50. Pearson WDS, Kulyk M, Kelly GM, Krone PH (1996) Cloning and characterization of a cDNA encoding the collagen-binding stress protein HSP47 in zebrafish. DNA Cell Biol 15(3):263–272CrossRefPubMedGoogle Scholar
  51. Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Res 29(9):e45CrossRefPubMedPubMedCentralGoogle Scholar
  52. Pirkkala L, Nykanen P, Sistonen L (2001) Roles of the heat shock transcription factors in regulation of the heat shock response and beyond. FASEB J 15:1118–1131CrossRefPubMedGoogle Scholar
  53. Podrabsky JE, Somero GN (2004) Changes in gene expression associated with acclimation to constant temperatures and fluctuating daily temperatures in an annual killifish Austrofundulus limnaeus. J Exp Biol 207:2237–2254CrossRefPubMedGoogle Scholar
  54. Purohit GK, Mahanty A, Suar M, Sharma AP, Mohanty BP, Mohanty S (2014) Investigating hsp gene expression in liver of channa striatus under heat stress for understanding the upper thermal acclimation. Biomed Res Int 2014:381719. doi: 10.1155/2014/381719 CrossRefPubMedPubMedCentralGoogle Scholar
  55. Purohit GK, Mahanty A, Mohanty BP, Mohanty S (2015) Evaluation of housekeeping genes as references for quantitative real-time PCR analysis of gene expression in the murrel Channa striatus under high-temperature stress. Fish Physiol Biochem. doi: 10.1007/s10695-015-0123-0 Google Scholar
  56. Reddy DV, Nagbhushanam P, Ramesh G (2013) Turnover time of Tural and Rajvadi hot spring waters, Maharashtra, India. Curr Sci 104(10):1419–1424Google Scholar
  57. Roberts R, Agius C, Saliba C, Bossier P, Sung Y (2010) Heat shock proteins (chaperones) in fish and shellfish and their potential role in relation to fish health: a review. J Fish Dis 33(10):789–801CrossRefPubMedGoogle Scholar
  58. Sanders BM, Martin LS, Wakagawa PA, Hunter DA, Miller S, Ullrich SJ (1994) Specific cross-reactivity of antibodies raised against two major stress proteins, stress 70 and chaperonin 60, in diverse species. Environ Toxicol Chem 13(8):1241–1249CrossRefGoogle Scholar
  59. Schuster-Bockler B, Conrad D, Bateman A (2010) Dosage sensitivity shapes the evolution of copy-number varied regions. PLoS ONE 5:e9474CrossRefPubMedPubMedCentralGoogle Scholar
  60. Shamovsky I, Nudler E (2008) New insights into the mechanism of heat shock response activation. Cell Mol Life Sci 65:855–861CrossRefPubMedGoogle Scholar
  61. Somero GN (2010) The physiology of climate change: how potentials for acclimatization and genetic adaptation will determine ‘winners’ and ‘losers’. J Exp Biol 213:912–920CrossRefPubMedGoogle Scholar
  62. Stone NM, Thomforde HK (2004) Understanding Your Fish Pond Water Analysis Report. Cooperative Extension Program, University of Arkansas at Pine Bluff Aquaculture/FisheriesGoogle Scholar
  63. Tedeschi JN, Kennington WJ, Berry O, Whiting S, Meekan M, Mitchell NJ (2015) Increased expression of Hsp70 and Hsp90 mRNA as biomarkers of thermal stress in loggerhead turtle embryos (Caretta Caretta). J Therm Biol 47:42–50CrossRefPubMedGoogle Scholar
  64. Tomanek L (2010) Variation in the heat shock response and its implication for predicting the effect of global climate change on species’ biogeographical distribution ranges and metabolic costs. J Exp Biol 213:971–979CrossRefPubMedGoogle Scholar
  65. Tomanek L, Somero GN (2002) Interspecific- and acclimation-induced variation in levels of heat-shock proteins 70 (hsp70) and 90 (hsp90) and heat-shock transcription factor-1 (HSF1) in congeneric marine snails (genus Tegula): implications for regulation of hsp gene expression. J Exp Biol 205(5):677–685PubMedGoogle Scholar
  66. Tosab M, Batten MR, Bulleid NJ (2000) Hsp47: a molecular chaperone that interacts with and stabilizes correctly-folded procollagen. EMBO J 19(10):2204–2211CrossRefGoogle Scholar
  67. Vihervaara A, Sistonen L (2014) HSF1 at a glance. J Cell Sci 127:261–266CrossRefPubMedGoogle Scholar
  68. Wang F, Dai AY, Tao K, Xiao Q, Huang ZL, Gao M, Li H, Wang X, Cao WX, Feng WL (2015) Heat shock protein-70 neutralizes apoptosis inducing factor in Bcr/Abl expressing cells. Cell Signal 27(10):1949–1955CrossRefPubMedGoogle Scholar
  69. Zeiner M, Cindric IJ, Pozgaj M, Pirkl R, Silic T, Stingeder G (2015) Influence of soil composition on the major, minor and trace metal content of Velebit biomedical plants. J Pharm Biomed Anal 106:153–158CrossRefPubMedGoogle Scholar
  70. Zunino B, Rubio-Patiño C, Villa E, Meynet O, Proics E, Cornille A, Pommier S, Mondragón L, Chiche J, Bereder J-M, Carles M, Ricci J-E (2016) Hyperthermic intraperitoneal chemotherapy leads to an anticancer immune response via exposure of cell surface heat shock protein 90. Oncogene 35:261–268CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Arabinda Mahanty
    • 1
    • 2
  • Gopal Krishna Purohit
    • 2
  • Ravi Prakash Yadav
    • 2
  • Sasmita Mohanty
    • 2
  • Bimal Prasanna Mohanty
    • 1
    Email author
  1. 1.Biochemistry Laboratory, Proteomics Unit, Fishery Resource and Environmental Management DivisionICAR- Central Inland Fisheries Research InstituteBarrackpore, KolkataIndia
  2. 2.KIIT School of BiotechnologyKIIT UniversityBhubaneswarIndia

Personalised recommendations