Differential expression and regulation of HSP70 gene during growth phase in ruminants in response to heat stress

Scientific Reports - Tập 12 Số 1
Rakesh Kaushik1, Anjana Goel1, P. K. Rout2
1Department of Biotechnology, GLA University, 17Km Stone, NH-2, Mathura-Delhi Road, Chaumuhan, Mathura, 281406, U.P, India
2Animal Genetics and Breeding Division, ICAR-Central Institute for Research on Goats, Makhdoom, Farah, Mathura, 281122, U.P, India

Tóm tắt

AbstractHeat shock proteins regulate the physiological mechanism of heat stress adaptation at cellular level. The present investigation was carried out to analyse the HSP70 gene regulation in various growth stage in ruminants in peripheral blood mononuclear cells (PBMCs). The relationship between HSP gene expression and thermotolerance in age-specific manner in ruminants has not been analysed. Therefore m-RNA HSP70 expression level was examined in different age groups of Jamunpari goat during hot climatic conditions. The experiment was carried out in 32 animals of Jamunapari goat belonging to the age groups of 3-months, 9-months, 12-months, and adults (2–3 year). Total RNA was isolated from peripheral blood mononuclear cells. The physiological response such as rectal temperature (RT), respiration rate (RR) and heart rate (HR) was used as indicator to heat stress. Temperature Humidity Index (THI) was used as an indicator of severity of environmental stress. The THI range varied from 82.00–92.08 during experimental period. The m-RNA HSP70 expression level at 9-month age of animals was up-regulated and significantly higher than other age groups. It was observed that the level of HSP70 transcripts in PBMCs was highest at 9-month age group, and age-related decline in HSP70 expression was observed in adult age. Based on the physiological response, the contrasting heat-stress phenotypes were recognised as heat stress susceptible (HSS) and heat stress tolerant (HST) individuals and the expression of m-RNA HSP70 was analysed at different ages in response to chronic heat stress. The differential mRNA expression of HSS individuals at 3 and 9-month of age showed the highest fold expression than HST. Age and phenotype had significant effect (p < 0.01) on the crossing point (CP) value. The m-RNA HSP70 gene expression in different age groups was correlated with heat stress tolerance and this could be used as biomarker for breeders to analyse the HSP responsein -vivoin ruminants.

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Tài liệu tham khảo

Carper, S. W., Duffy, J. J. & Gerner, E. W. Heat shock proteins in thermotolerance and other cellular processes. Can. Res. 47(20), 5249–5255 (1987).

Kregel, K. C. Invited review—Heat shock proteins: Modifying factors in physiological stress responses and acquired thermotolerance. J. Appl. Physiol. 92, 2177–2186 (2002).

Calderwood, S. K., Murshid, A. & Prince, T. The shock of aging: Molecular chaperones and the heat shock response in longevity and aging—A mini-review. Gerontology 55(5), 550–558 (2009).

Kim, B., Park, K. & Rhee, K. Heat stress response of male germ cells. Cell. Mol. Life Sci. 70(15), 2623–2636 (2013).

Sorger, P. K. The transcriptional regulation of heat shock genes (Doctoral dissertation) (University of Cambridge, Cambridge, 1987).

Sorger, P. K., Lewis, M. J. & Pelham, H. R. Heat shock factor is regulated differently in yeast and HeLa cells. Nature 329(6134), 81–84 (1987).

Zimarino, V. & Wu, C. Induction of sequence-specific binding of Drosophila heat shock activator protein without protein synthesis. Nature 327(6124), 727–730 (1987).

Lindquist, S. & Craig, E. A. The heat-shock proteins. Annu. Rev. Genet 22, 631–677 (1988).

Sonna, L. A., Fujita, J., Gaffin, S. L. & Lilly, C. M. Invited review: Effects of heat and cold stress on mammalian gene expression. J. Appl. Physiol. 92(4), 1725–1742 (2002).

Morange, F. HSFs in development. Handb. Exp. Pharmacol. 172, 153–169 (2006).

Fargnoli, J., Kunisada, T., Fornace, A. J., Schneider, E. L. & Holbrook, N. J. Decreased expression of heat shock protein 70 mRNA and protein after heat treatment in cells of aged rats. Proc. Natl. Acad. Sci. 87(2), 846–850 (1990).

Finch, C. E., Landfield, P. W. in Handbook of the Biology of Aging (eds. Finch, C. E. & Schneider, E. L.) (Van Nostrand Reinhold, New York, 1978) pp. 567–594.

Barbe, M. F., Tytell, M., Gower, D. J. & Welch, W. J. Hyperthermia protects against light damage in the rat retina. Science 241(4874), 1817–1820 (1988).

Rout, P. K., Kaushik, R., Ramachandran, N. & Jindal, S. K. Identification of heat stress-susceptible and-tolerant phenotypes in goats in semiarid tropics. Animal Production Science 58(7), 1349–1357 (2017).

Andrews, A. H. Veterinary medicine. In A textbook of the Diseases of Cattle Sheep Pigs Goats and Horses (eds Blood, D. C. & Radostits, O. M.) 1502 (Baillière Tindall, London, 1988).

McDowell, R. E., Hooven, N. W. & Camoens, J. K. Effect of climate on performance of Holsteins in first lactation. J. Dairy Sci. 59(5), 965–997 (1976).

Rout, P. K., Kaushik, R. & Ramachandran, N. Differential expression pattern of heat shock protein 70 gene in tissues and heat stress phenotypes in goats during peak heat stress period. Cell Stress Chaperones 21(4), 645–651 (2016).

Kaushik, R., Dige, M. S. & Rout, P. K. Molecular characterization and expression profiling of ENOX2 gene in response to heat stress in goats. Cell Dev. Biol 5, 1–5 (2016).

Kaushik, R., Dige, M., Dass, G., Ramachandran, N. & Rout, P. K. Superoxide dismutase activity in response to heat stress in Jamunapari goats. Indian J. Biochem. Biophys. 55(1), 39–43 (2018).

Kaushik, R., Goel, A. & Rout, P. K. Differential expression and characterization of ATP1A1 exon17 gene by high resolution melting analysis and RT-PCR in Indian goats. Mol. Biol. Rep. 46(5), 5273–5286 (2019).

Kaushik, R., Goel, A. & Rout, P. K. Establishing the genetic variation in physiological response in response to heat stress in semi-arid region in Jamunapari goats. Biol. Rhythm. Res. 51(1), 1–14 (2020).

Livak, K. J. & Schmittgen, T. D. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25(4), 402–408 (2001).

Harvey, W. R. User's Guide for LSMLMW In: Mixed model least Squares and Maximum Likelihood Computer Program PC-Version, Vol. 2, (Ohio State University Press, Ohio, 1990).

Beere, H. M. et al. Heat-shock protein 70 inhibits apoptosis by preventing recruitment of procaspase-9 to the Apaf-1 apoptosome. Nat. Cell Biol. 2(8), 469–475 (2000).

Sonna, L. A., Fujita, J., Gaffin, S. L. & Lilly, C. M. Invited review: Effects of heat and cold stress on mammalian gene expression. J. Appl. Physiol. 92(4), 1725–1742 (1985).

Hecker, J. G. et al. Heat shock proteins HSP70 and HSP27 in the cerebral spinal fluid of patients undergoing thoracic aneurysm repair correlate with the probability of postoperative paralysis. Cell Stress Chaperones 13(4), 435–446 (2008).

Hecker, J. G. & McGarvey, M. Heat shock proteins as biomarkers for the rapid detection of brain and spinal cord ischemia: A review and comparison to other methods of detection in thoracic aneurysm repair. Cell Stress Chaperones 16(2), 119–131 (2011).

Archana, P. R. et al. Role of heat shock proteins in livestock adaptation to heat stress. J. Dairy Vet. Anim. Res. 5(1), 00127 (2017).

Hyder, I. et al. Thermotolerance in domestic ruminants: A HSP70 perspective. In Heat Shock Proteins in Veterinary Medicine and Sciences (eds Alexzander, A. A. & Asea, P. K.) 3–35 (Springer, Cham, 2017).

Pacifici, M. et al. Generation length for mammals. Nat. Conserv. 13(5), 84–89 (2013).

Webb, E. C. & Casey, N. H. Physiological limits to growth and the related effects on meat quality. Livest. Sci. 130(1–3), 33–40 (2010).

Yilmaz, M., Kayki, M., Aşici, G. & Kiral, F. The mRNA gene expression profiles for HSP60 and HSP70 in various aged Saanen goats in different seasons. Kafkas Üniv. Vet. Fak. Derg. 24(1), 563–569 (2018).

Alyamani, D., Koluman, N. (2019) Associated expressions of heat shock protein (70 and 60) with physiological adaptation with in dairy goats. Available at SSRN 3535801.

Alyamani, D. Impact various seasons on expression patterns HSP60 and physiological parameters. J Dairy Vet Anim Res 9(1), 1–4 (2020).

Heydari, A. R., Wu, B., Takahashi, R. Y. O. Y. A., Strong, R. A. N. D. Y. & Richardson, A. Expression of heat shock protein 70 is altered by age and diet at the level of transcription. Mol. Cell. Biol. 13(5), 2909–2918 (1993).

Yamani, H. A. L. & Koluman, N. Association HSP70 with some physiological parameters in dairy goat under south Turkey conditions. J. Dairy. Vet. Anim. Res. 9(5), 148–151 (2020).

Pavlov, E. P., Harman, S. M., Chrousos, G. P., Loriaux, D. L. & Blackman, M. R. Responses of plasma adrenocorticotropin, cortisol, and dehydroepiandrosterone to ovine corticotropin-releasing hormone in healthy aging men. J. Clin. Endocrinol. Metab. 62(4), 767–772 (1986).

Roth, G. Brain Res. 107, 354–349 (1976).

Brodish, A. & Odio, M. Age-dependent effects of chronic stress on ACTH and corticosterone responses to an acute novel stress. Neuroendocrinology 49(5), 496–501 (1989).

Ottenweller, J. E., Tapp, W. N., Pitman, D. L. & Natelson, B. H. Interactions among the effects of aging, chronic disease, and stress on adrenocortical function in Syrian hamsters. Endocrinology 126(1), 102–109 (1990).

Landfield, P. W., Waymire, J. C. & Lynch, G. Hippocampal aging and adrenocorticoids: Quantitative correlations. Science 202(4372), 1098–1102 (1978).

Sapolsky, R. M., Krey, L. C. & McEWEN, B. S. Corticosterone receptors decline in a site-specific manner in the aged rat brain. Brain Res. 289(1–2), 235–240 (1983).

Sapolsky, R. M., Krey, L. C. & McEwen, B. S. The neuroendocrinology of stress and aging: The glucocorticoid cascade hypothesis. Endocr. Rev. 7, 284–297 (1986).

Odio, M. & Brodish, A. Age-related adaptation of pituitary-adrenocortical responses to stress. Neuroendocrinology 49(4), 382–388 (1989).

Finley, D., Ciechanover, A. & Varshavsky, A. Thermolability of ubiquitin-activating enzyme from the mammalian cell cycle mutant ts85. Cell 37(1), 43–55 (1984).

Gershon, H. & Gershon, D. Inactive enzyme molecules in aging mice: Liver aldolase. Proc. Natl. Acad. Sci. 70(3), 909–913 (1973).

Liu, A. Y., Lin, Z., Choi, H. S., Sorhage, F. & Li, B. Attenuated induction of heat shock gene expression in aging diploid fibroblasts. J. Biol. Chem. 264(20), 12037–12045 (1989).

Deguchi, Y., Negoro, S. & Kishimoto, S. Molecular chaperones and the aging process Biochem. Biophys. Res. Commun. 157, 580–584 (1998).

Jin, X. et al. Serum and lymphocyte levels of heat shock protein 70 in aging: A study in the normal Chinese population. Cell Stress Chaperones 9(1), 69 (2004).