Abstract
Six male Tharparkar cattle of 2–3 years old were selected for the study. After 15-day acclimation at thermoneutral zone (TNZ) in psychrometric chamber, animals were exposed at 42 °C for 6 h up to 23 days followed by 12 days of recovery period. Blood samples were collected during control period at TNZ (days 1, 5, and 12), after heat stress exposure (day 1, immediate heat stress acclimation (IHSA); days 2 to 10, short-term heat stress acclimation (STHSA); days 15 to 23, long-term heat stress acclimation (LTHSA); days 7 and 12, recovery period), and peripheral blood mononuclear cells (PBMCs) were isolated for RNA and protein extraction. The messenger RNA (mRNA) and protein expression in PBMCs were determined by qPCR and western blot, respectively. Samples at TNZ were taken as control. The mRNA expression of HSP90, iNOS, and eNOS was significantly upregulated (P < 0.05) on day 1 (ISHA) as compared to control, remained consistent during STHSA, again increased during LTHSA, and finally reduced to basal level during recovery period. The protein expression of HSP90, iNOS, and eNOS were akin to their transcript pattern. PBMC culture study was conducted to study transcriptional abundance of HSP90, iNOS, and eNOS at different temperature-time combinations. The present findings indicate that HSP90, iNOS, and eNOS could possibly play an important role in mitigating thermal insults and confer thermotolerance during long-term heat stress exposure in Tharparkar cattle.
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Alderton WK, Cooper CE, Knowles RG (2001) Nitric oxide synthases: structure, function and inhibition. Biochem J 357:593–615
Ali EMM, Soha HM, Mohamed TM (2012) Nitric oxide synthase and oxidative stress: regulation of nitric oxide synthase. In: Volodymyr Lushchak (ed.) Oxidative stress-molecular mechanisms and biological effects. InTech, 61–72 ISBN: 978–953–51-0554-1
Arnaud C, Laubriet A, Joyeux M, Godin-Ribuot D, Rochette L, Demenge P, Ribuot C (2001) Role of nitric oxide synthases in the infarct size-reducing effect conferred by heat stress in isolated rat hearts. Br J Pharmacol 132:1845–1851
Aschner JL, Foster SL, Kaplowitz M, Zhang Y, Zeng H, Fike CD (2007) Heat shock protein 90 modulates endothelial nitric oxide synthase activity and vascular reactivity in the newborn piglet pulmonary circulation. Am. J. Physiol. Lung Cell Mol Physiol 292:L1515–L1525
Billecke SS, Bender AT, Kanelakis KC, Murphy PJ, Lowe ER, Kamada Y, Pratt WB, Osawa Y (2002) hsp90 is required for heme binding and activation of aponeuronal nitric-oxide synthase: geldanamycin-mediated oxidant generation is unrelated to any action of hsp90. J Biol Chem 277:20504–20509
Bogdan C (2001) Nitric oxide and the immune response. Nat Immunol 2:907–916
Coleman JW (2001) Nitric oxide in immunity and inflammation. Int Immunopharmacol 1:1397–1406
Collier RJ, Dahl GE, Van Baale MJ (2006) Major advances associated with environmental effects on dairy cattle. J Dairy Sci 89:1244–1253
Csermely P, Schnaider T, Soti C, Prohaszka Z, Nardai G (1998) The 90-kDa molecular chaperone family: structure, function and clinical applications: a comprehensive review. Pharmacol Ther 79:129–168
Dangi SS, Dangi SK, Chouhan VS, Verma MR, Kumar P, Singh G, Sarkar M (2016) Modulatory effect of betaine on expression dynamics of HSPs during heat stress acclimation in goat (Capra hircus). Gene 575:543–550
Dangi SS, Gupta M, Maurya D, Yadav VP, Panda RP, Singh G, Mohan NH, Bhure SK, Das BC, Bag S, Mahapatra R, Sharma GT, Sarkar M (2012) Expression profile of HSP genes during different seasons in goats (Capra hircus). Trop Anim Health Prod 44:1905–1912
Dangi SS, Gupta M, Nagar V, Yadav VP, Dangi SK, Om S, Chouhan VS, Kumar P, Singh G, Sarkar M (2014) Impact of short-term heat stress on physiological responses and expression profile of HSPs in Barbari goats. Int J Biometeorol 58:2085–2093
Dubey PK, Tripathi V, Singh RP, Saikumar G, Nath A, Pratheesh GN, Sharma GT (2012) Expression of nitric oxide synthase isoforms in different stages of buffalo (Bubalus bubalis) ovarian follicles: effect of nitric oxide on in vitro development of preantral follicle. Theriogenology 77:280–291
Duval M, Boeuf FL, Huot J, Gratton JP (2007) Src-mediated phosphorylation of Hsp90 in response to vascular endothelial growth factor (VEGF) is required for VEGF receptor-2 signaling to endothelial NO synthase. Mol Biol Cell 18:4659–4668
Garcia-Cardena G, Fan R, Shah V, Sorrentino R, Cirino G, Papapetropoulos A, Sessa WC (1998) Dynamic activation of endothelial nitric oxide synthase by Hsp90. Nature 392:821–824
Govers R, van der Sluijs P, van Donselaar E, Slot J, Rabelink TJ (2002) Endothelial nitric oxide synthase and its negative regulator caveolin-1 localize to distinct perinuclear organelles. J Histochem Cytochem 50:779–788
Gratton JP, Fontana J, O’Connor DS, Garcia-Cardena G, McCabe TJ, Sessa WC (2000) Reconstitution of an endothelial nitric-oxide synthase (eNOS), hsp90, and caveolin-1 complex in vitro. Evidence that hsp90 facilitates calmodulin stimulated displacement of eNOS from caveolin-1. J Biol Chem 275:22268–22272
Kellogg D, Crandall C, Liu Y, Charkoudian N, Johnson J (1998) Nitric oxide and cutaneous active vasodilation during heat stress in humans. J Appl Physiol 85:824–829
Kishore A, Sodhi M, Kumari P, Mohanty AK, Sadana DK, Kapila N, Khate K, Shandilya U, Kataria RS, Mukesh M (2014) Peripheral blood mononuclear cells: a potential cellular system to understand differential heat shock response across native cattle (Bos indicus), exotic cattle (Bos taurus), and riverine buffaloes (Bubalus bubalis) of India. Cell Stress Chaperones 19:613–621
Lee JH, Yang ES, Park JW (2003) Inactivation of NADP+-dependent isocitrate dehydrogenase by peroxynitrite. Implications for cytotoxicity and alcohol-induced liver injury. J Biol Chem 278:51360–51371
Li Z, Srivastava P (2004) Heat-shock proteins Current protocols in immunology / edited by John E Coligan [et al.] Appendix 1: Appendix 1T doi:10.1002/0471142735.ima01ts58
Lindquist S, Craig EA (1988) The heat-shock proteins. Annu Rev Genet 22:631–677
Luo S, Wang T, Qin H, Lei H, Xia Y (2011) Obligatory role of heat shock protein 90 in iNOS induction. Am J Physiol Cell Physiol 301:C227–C233
Mehla K, Magotra A, Choudhary J, Singh AK, Mohanty AK, Upadhyay RC, Srinivasan S, Gupta P, Choudhary N, Antony B, Khan F (2014) Genome-wide analysis of the heat stress response in zebu (sahiwal) cattle. Gene 533:500–507
Mishra A, Hooda OK, Singh G, Meur SK (2010) Influence of induced heat stress on HSP70 in buffalo lymphocytes. J Anim Physiol Anim Nutr 95:540–544
Mishra SR, Thakur N, Somal A, Parmar MS, Yadav VP, Bharati J, Bharti MK, Paul A, Verma MR, Chouhan VS, Sharma GT, Singh G, Gonzalez LA, D’Occhio MJ, Sarkar M (2016) Expression and localization of angiopoietin family in buffalo ovarian follicles during different stages of development and modulatory role of angiopoietins on steroidogenesis and survival of cultured buffalo granulosa cells. Theriogenology 86:1818–1833
Mori M (2007) Regulation of nitric oxide synthesis and apoptosis by arginase and arginine recycling. J Nutr 137:1616S–1620S
Nathan C (1997) Inducible nitric oxide synthase: what difference does it make? J Clin Invest 100:2417–2423
Pearl LH, Prodromou C, Workman P (2008) The Hsp90 molecular chaperone: an open and shut case for treatment. Biochem J 410:439–453
Pfaffl M (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:2002–2007
Pritchard KA Jr, Ackerman AW, Gross ER, Stepp DW, Shi Y, Fontana JT, Baker JE, Sessa WC (2001) J Biol Chem 276:17621–17624
Sharma HS, Drieu K, Alm P, Westman J (2000) Role of nitric oxide in blood-brain barrier permeability, brain edema and cell damage following hyperthermic brain injury. An experimental study using EGB-761 and Gingkolide B pretreatment in the rat. In: Brain edema XI. Springer, Vienna pp, pp 81–86
Silanikove N (1992) Effects of water scarcity and hot environment on appetite and digestion in ruminants: a review. Livest Prod Sci 30:175–194
Silanikove N (2000) Effects of heat stress on the welfare of extensively managed domestic ruminants. Livest Prod Sci 67:1–18
Song Y, Zweier JL, Xia Y (2001) Heat-shock protein 90 augments neuronal nitric oxide synthase activity by enhancing Ca2+/calmodulin binding. Biochem J 355:357–360
Sonna LA, Gaffin SL, Pratt RE, Cullivan ML, Angel KG, Lilly CM (2002) Effects of acute heat shock on gene expression by human peripheral blood mononuclear cells. J Appl Physiol 92:2208–2220
Xu H, Shi Y, Wang J, Jones D, Weilrauch D, Ying R, Wakim B, Pritchard KA Jr (2007) A heat shock protein 90 binding domain in endothelial nitric-oxide synthase influences enzyme function. J Biol Chem 282:37567–37574
Yadav VP, Dangi SS, Chouhan VS, Gupta M, Dangi SK, Singh G, Maurya VP, Kumar P, Sarkar M (2016) Expression analysis of NOS family and HSP genes during thermal stress in goat (Capra hircus). Int J Biometeorol 60:381–389
Yoshida M, Xia Y (2003) Heat shock protein 90 as an endogenous protein enhancer of inducible nitric-oxide synthase. J Biol Chem 278:36953–36958
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We thank National Initiative on Climate Resilient Agriculture (NICRA) and Director, Indian Veterinary Research Institute, Izatnagar, India, for funding of this work.
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Bharati, J., Dangi, S., Bag, S. et al. Expression dynamics of HSP90 and nitric oxide synthase (NOS) isoforms during heat stress acclimation in Tharparkar cattle. Int J Biometeorol 61, 1461–1469 (2017). https://doi.org/10.1007/s00484-017-1323-3
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DOI: https://doi.org/10.1007/s00484-017-1323-3