Acta Scientific Microbiology (ISSN: 2581-3226)

Research Article Volume 5 Issue 11

Comparative Analysis of Heat Shock Protein Genes Hsp70 and Hsp90 Expression by Live and Killed Edwardsiella tarda Challenge in Rohu, Labeo rohita

Akash Jayalakshmi Subramaniyan, Kurcheti Pani Prasad, Saurav Kumar, Nalini Poojary, Anisha Valsalam and Megha Kadam Bedekar*

Aquatic Environment and Health Management Division, ICAR-Central Institute of Fisheries Education, Mumbai, India

*Corresponding Author: Megha Kadam Bedekar, Aquatic Environment and Health Management Division, ICAR-Central Institute of Fisheries Education, Mumbai, India.

Received: October 03, 2022; Published: October 21, 2022

Abstract

An inclusive variety of biotic and abiotic pressures fuels specific intracellular stress responses characterized by a family of unique proteins referred to as heat shock proteins (HSP). The present study is an expression profiling of two significant heat shock protein genes, Hsp70 and Hsp90, during live (lethal, sublethal) and killed Edwardsiella tarda challenge in rohu, Labeo rohita. Thirty rohu juveniles were challenged with the lethal dose (LD50) of E. tarda 3.5×107 CFU, the sublethal dose of E. tarda 3.5×106 CFU, and formalin-killed E. tarda 3.5×107 CFU per 20g of body weight of fish, intraperitoneally. The genes' (Hsp70 and Hsp90) expressions were studied in the liver, kidney, spleen, and gill tissues of rohu at different periods: 0, 3, 6, 12, 24, 48, and 72 hours post-challenge by qPCR. The Hsp gene modulation was more significant in the liver and spleen than in kidney and gill tissues. Among the three treatments, the killed E. tarda showed more expression of Hsp70 production followed by the lethal dose. High Hsp90 expression was observed in fishes challenged with live E. tarda than killed bacteria. Further, the highest activity of glutathione peroxidase and myeloperoxidase was found in all the treatments. The study indicated the critical role of Hsp70 and Hsp90 in live infection compared to killed bacterial administration.

Keywords: Heat Shock Proteins; HSP 70; HSP 90; Edwardsiella tarda; Labeo rohita

References

  1. Sanders Brenda M. "Stress proteins in aquatic organisms: an environmental perspective”. Critical reviews in Toxicology1 (1993): 49-75.
  2. Ritossa Ferruccio. "A new puffing pattern induced by temperature shock and DNP in Drosophila”. Experientia12 (1962): 571-573.
  3. Finlayson-Trick Emma., et al. "Regulation of antimicrobial pathways by endogenous heat shock proteins in gastrointestinal disorders”. Gastrointestinal Disorders 1 (2018): 39-56.
  4. Basu N., et al. "Heat shock protein genes and their functional significance in fish”. Gene2 (2002): 173-183.
  5. Gupta Radhey S and G. Brian Golding. "Evolution of HSP70 gene and its implications regarding relationships between archaebacteria, eubacteria, and eukaryotes”. Journal of Molecular Evolution 6 (1993): 573-582.
  6. Ali Adnan., et al. "Isolation and characterization of a cDNA encoding a Xenopus 70-kDa heat shock cognate protein, Hsc70. I”. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 4 (1996): 681-687.
  7. Brodsky Jeffrey L and Gabriela Chiosis. "Hsp70 molecular chaperones: emerging roles in human disease and identification of small molecule modulators”. Current Topics in Medicinal Chemistry11 (2006): 1215-1225.
  8. Wadhwa Renu., et al. "An Hsp70 family chaperone, mortalin/mthsp70/PBP74/Grp75: what, when, and where?”. Cell Stress and Chaperones3 (2002): 309.
  9. Marchesi VT and Nhu Ngo. "In vitro assembly of multiprotein complexes containing alpha, beta, and gamma tubulin, heat shock protein HSP70, and elongation factor 1 alpha”. Proceedings of the National Academy of Sciences7 (1993): 3028-3032.
  10. Liu Zi-Ming., et al. "Effect of high temperature stress on heat shock protein expression and antioxidant enzyme activity of two morphs of the mud crab Scylla paramamosain”. Comparative Biochemistry and Physiology Part A: Molecular and Integrative Physiology 223 (2018): 10-17.
  11. Mazur Carl François. “The heat shock protein response and physiological stress in aquatic organisms”. Diss. University of British Columbia, (1996).
  12. Das Sweta., et al. "Expression analysis of heat shock protein genes during Aeromonas hydrophila infection in rohu, Labeo rohita, with special reference to molecular characterization of Grp78”. Cell Stress and Chaperones1 (2015): 73-84.
  13. Donzé Olivier and Didier Picard. "Hsp90 binds and regulates the ligand-inducible α subunit of eukaryotic translation initiation factor kinase Gcn2”. Molecular and Cellular Biology12 (1999): 8422-8432.
  14. Smith David F., et al. "Identification of a 60-kilodalton stress-related protein, p60, which interacts with hsp90 and hsp70”. Molecular and Cellular Biology2 (1993): 869-876.
  15. Caplan Avrom J. "Hsp90's secrets unfold: new insights from structural and functional studies”. Trends in Cell Biology 7 (1999): 262-268.
  16. Palmisano Aldo N., et al. "Tissue-specific induction of Hsp90 mRNA and plasma cortisol response in chinook salmon following heat shock, seawater challenge, and handling challenge”. Marine Biotechnology 4 (2000): 329-338.
  17. Forsyth RB., et al. "Stress protein expression in coho salmon with bacterial kidney disease”. Journal of Aquatic Animal Health1 (1997): 18-25.
  18. Roberts R J., et al. "Heat shock proteins (chaperones) in fish and shellfish and their potential role in relation to fish health: a review”. Journal of Fish Diseases10 (2010): 789-801.
  19. FARMER III JJ. "Genus X. Edwardsiella”. Bergey's Manual of Systematic Bacteriology 1 (1984): 486-491.
  20. Janda JM., et al. "Pathogenic properties of Edwardsiella species”. Journal of Clinical Microbiology9 (1991): 1997-2001.
  21. Lowry Toby and Stephen A. Smith. "Aquatic zoonoses associated with food, bait, ornamental, and tropical fish”. Journal of the American Veterinary Medical Association6 (2007): 876-880.
  22. Swain P and S K Nayak. "Comparative sensitivity of different serological tests for seromonitoring and surveillance of Edwardsiella tarda infection of Indian major carps”. Fish and Shellfish Immunology4 (2003): 333-340.
  23. Du Yang., et al. "The influence of concentration of inactivated Edwardsiella tarda bacterin and immersion time on antigen uptake and expression of immune-related genes in Japanese flounder (Paralichthys olivaceus)”. Microbial Pathogenesis 103 (2017): 19-28.
  24. Reed Lowell Jacob and Hugo Muench. "A simple method of estimating fifty per cent endpoints”. American Journal of Epidemiology3 (1938): 493-497.
  25. Robinson Nicholas., et al. "Expressed sequences and polymorphisms in rohu carp (Labeo rohita, Hamilton) revealed by mRNA-seq”. Marine Biotechnology5 (2012): 620-633.
  26. Livak Kenneth J and Thomas D Schmittgen. "Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method”. Methods4 (2001): 402-408.
  27. Sahoo P K., et al. “Non‐specific immune responses in juveniles of Indian major carps”. Journal of Applied Ichthyology 2 (2005): 151-155.
  28. Lawrence Richard A and Raymond F Burk. "Glutathione peroxidase activity in selenium-deficient rat liver”. Biochemical and Biophysical Research Communications 4 (1976): 952-958.
  29. Parsell D A and Susan Lindquist. "The function of heat-shock proteins in stress tolerance: degradation and reactivation of damaged proteins”. Annual Review Of Genetics 27 (1993): 437-497.
  30. Yang J., et al. “Pathogenic infection and immune-related gene expression of Chinese sturgeon (Acipenser sinensis) challenged by Citrobacter freundii”. Developmental and Comparative Immunology 114 (2021): 103872.
  31. Bosch TC., et al. "Thermotolerance and synthesis of heat shock proteins: these responses are present in Hydra attenuata but absent in Hydra oligactis”. Proceedings of the National Academy of Sciences21 (1988): 7927-7931.
  32. Sanders B M and L S Martin. "Stress proteins as biomarkers of contaminant exposure in archived environmental samples”. Science of the Total Environment 139 (1993): 459-470.
  33. Dietz Thomas J and George N Somero. "Species-and tissue-specific synthesis patterns for heat-shock proteins HSP70 and HSP90 in several marine teleost fishes”. Physiological Zoology6 (1993): 863-880.
  34. Nakano T., et al. “Effect of bacterial infection on the expression of stress proteins and antioxidative enzymes in Japanese flounder”. In Evolution of Marine Coastal Ecosystems under the Pressure of Global Changes” (2020): 111-127.
  35. Nasrullah Hasan., et al. "Early immune gene development and expression in African catfish Clarias gariepinus after challenged with Aeromonas hydrophila”. Aquaculture International2 (2021): 595-607.
  36. Wang Pengfei., et al. "Identification and expression analysis of two HSP70 isoforms in mandarin fish Siniperca chuatsi”. Fisheries science4 (2014): 803-817.
  37. Ackerman Paige A and George K Iwama. "Physiological and cellular stress responses of juvenile rainbow trout to vibriosis”. Journal of Aquatic Animal Health2 (2001): 173-180.
  38. Sung Y Yik and T H MacRae. "Heat shock proteins and disease control in aquatic organisms”. Journal of Aquaculture Research and Development S 2.006 (2011).
  39. Peng Shuang., et al. "Heat shock protein 70 gene of ya-fish (Schizothorax prenanti) responses to thermal stress and bacterial challenges”. (2018).
  40. Xie Yangjie., et al. "Hsp90, Hsp60 and sHsp families of heat shock protein genes in channel catfish and their expression after bacterial infections”. Fish and Shellfish Immunology2 (2015): 642-651.
  41. Dietz Thomas J and George N Somero. "The threshold induction temperature of the 90-kDa heat shock protein is subject to acclimatization in eurythermal goby fishes (genus Gillichthys)”. Proceedings of the National Academy of Sciences8 (1992): 3389-3393.
  42. Chen Young-Mao., et al. "Cloning of an orange-spotted grouper Epinephelus coioides heat shock protein 90AB (HSP90AB) and characterization of its expression in response to nodavirus”. Fish and shellfish immunology5-6 (2010): 895-904.
  43. Lee Joo-Yang., et al. "Monoclonal antibodies raised against infectious haematopoietic necrosis virus (IHNV) G protein and a cellular 90 kDa protein neutralize IHNV infection in vitro”. Journal of general Virology8 (1996): 1731-1737.
  44. Li Juan., et al. "Gill transcriptomes reveal expression changes of genes related with immune and ion transport under salinity stress in silvery pomfret (Pampus argenteus)”. Fish Physiology and Biochemistry4 (2020): 1255-1277.
  45. Rungrassamee Wanilada., et al. "Expression and distribution of three heat shock protein genes under heat shock stress and under exposure to Vibrio harveyi in Penaeus monodon”. Developmental and Comparative Immunology10 (2010): 1082-1089.
  46. Zhou Aiguo., et al. "Molecular cloning, characterization and expression analysis of heat shock protein 90 in albino northern snakehead Channa argus”. Gene 626 (2017): 173-181.
  47. Haug Gerd., et al. "The host cell chaperone Hsp90 is necessary for cytotoxic action of the binary iota-like toxins”. Infection and Immunity5 (2004): 3066-3068.
  48. Vojtech Lucia N., et al. "Host immune response and acute disease in a zebrafish model of Francisella pathogenesis”. Infection and Immunity2 (2009): 914-925.
  49. Pu Yundan., et al. "Molecular characterization and expression analysis of Hsp90 in Schizothorax prenanti”. Cell Stress and Chaperones6 (2016): 983-991.
  50. Klebanoff Seymour J. "Myeloperoxidase: friend and foe”. Journal of Leukocyte Biology5 (2005): 598-625.
  51. Stosik M., et al. "Resistance in carps (Cyprinus carpio) affected by a natural bacterial infection”. VETERINARNI MEDICINA-PRAHA- 46.1 (2001): 6-15.
  52. Sahoo P K., et al. "Selection for improved resistance to Aeromonas hydrophila in Indian major carp Labeo rohita: Survival and innate immune responses in first generation of resistant and susceptible lines”. Fish and Shellfish Immunology3 (2011): 432-438.
  53. Behera Truptimayee and Priyabrat Swain. "Antigen adsorbed calcium phosphate nanoparticles stimulate both innate and adaptive immune response in fish, Labeo rohita H”. Cellular Immunology2 (2011): 350-359.
  54. Mohanty B R and P K Sahoo. "Immune responses and expression profiles of some immune-related genes in Indian major carp, Labeo rohita to Edwardsiella tarda infection”. Fish and Shellfish Immunology4 (2010): 613-621.
  55. Zhang Linbao., et al. "Transcriptional regulation of selenium-dependent glutathione peroxidase from Venerupis philippinarum in response to pathogen and contaminants challenge”. Fish and Shellfish Immunology6 (2011): 831-837.

Citation

Citation: Megha Kadam Bedekar., et al. “Comparative Analysis of Heat Shock Protein Genes Hsp70 and Hsp90 Expression by Live and Killed Edwardsiella tarda Challenge in Rohu, Labeo rohita". Acta Scientific Microbiology 5.11 (2022): 57-67.

Copyright

Copyright: © 2022 Megha Kadam Bedekar., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.




Metrics

Acceptance rate30%
Acceptance to publication20-30 days

Indexed In






News and Events


  • Certification for Review
    Acta Scientific certifies the Editors/reviewers for their review done towards the assigned articles of the respective journals.
  • Submission Timeline for Upcoming Issue
    The last date for submission of articles for regular Issues is July 10, 2024.
  • Publication Certificate
    Authors will be issued a "Publication Certificate" as a mark of appreciation for publishing their work.
  • Best Article of the Issue
    The Editors will elect one Best Article after each issue release. The authors of this article will be provided with a certificate of "Best Article of the Issue"
  • Welcoming Article Submission
    Acta Scientific delightfully welcomes active researchers for submission of articles towards the upcoming issue of respective journals.

Contact US