Skip to main content
Log in

Cellular thermotolerance is associated with heat shock protein 70.1 genetic polymorphisms in Holstein lactating cows

  • Original Paper
  • Published:
Cell Stress and Chaperones Aims and scope

Abstract

Heat shock proteins (Hsp) are known to protect cells from several stressors. Nucleotide changes in the flanking regions [5′- and 3′-untranslated region (UTR)] of Hsp gene might affect inducibility, degree of expression, or stability of Hsp70 mRNA. The present study aimed to investigate the association between inducible Hsp70.1 single nucleotide polymorphisms (SNPs) and heat shock (HS) response of peripheral blood mononuclear cells (PBMC) in dairy cows. Four hundred forty-six Italian Holstein cows were genotyped for four Hsp70.1 SNPs: g895 C/- and g1128 G/T in 5′-UTR, and g2154 G/A and g64 G/T in 3′-UTR. Genetic polymorphisms in 3′-UTR of bovine Hsp70.1 gene resulted monomorphic. Distribution of alleles of the nucleotide sequence polymorphism within the 5′-UTR of the bovine Hsp70.1 gene were 81.2% and 18.8% for C and -, respectively, and 77.8% and 22.2% for G and T, respectively. Among the 446 genotyped animals, a group of cows balanced for days in milk and parity was selected to be representative of the following genotypes: CC (n = 8), C- (n = 7), and -- (n = 7) and GG (n = 8), GT (n = 11), and TT (n = 3) in 5′-UTR. PBMC were isolated from blood samples and heated at 43°C in thermal bath for 1 h and then incubated at 39°C in atmosphere of 5% CO2 for 1, 2, 4, 8, 16, and 24 h (recovery times). Cell viability was determined by XTT assay. Gene and protein expression of Hsp70.1 was determined by real-time reverse transcription-polymerase chain reaction and by ELISA assay, respectively. For the two SNPs detected, one allele was the most frequent (C, 66.8% and G, 56.8%). Genotypes -- and TG showed higher (P < 0.05) viability compared with CC and GG, respectively. Genotypes C- and TT had intermediate viability. Gene expression of Hsp70.1 showed higher (P < 0.001) levels in -- and TG genotype compared with their counterparts. Genotypes -- and TG showed the higher level of inducible Hsp70.1 protein in respect to C-, TT and CC, GG. In conclusion, exposure to HS differently affected cell viability and gene and protein expression of Hsp70.1 in the selected genotypes. These results indicate that the presence of SNPs (C/- and G/T) in the 5′-UTR region of inducible Hsp70.1 ameliorates HS response and tolerance to heat of bovine PBMC. These mutation sites may be useful as molecular genetic markers to assist selection for heat tolerance.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Adamowicz T, Pers E, Lechniak D (2005) A new SNP in the 3′-UTR of the Hsp70-1 Gene in Bos taurus and Bos indicus. Biochem Genet 43:623–627

    Article  PubMed  CAS  Google Scholar 

  • Banks A, Looper ML, Reiter S, Starkey L, Flores R, Hallford D, Rosenkrans C Jr (2007) Identification of single nucleotide polymorphisms within the promoter region of the bovine heat shock protein 70 gene and associations with pregnancy. Am Soc Anim Sci South Sect Meet 85(suppl 2):10

    Google Scholar 

  • Beckham JT, Mackanos MA, Crooke C, Takahashi T, O'Connell-Rodwell C, Contag CH, Jansen ED (2004) Assessment of cellular response to thermal laser injury through bioluminescence imaging of heat shock protein 70. Photochem Photobiol 79:76–85

    PubMed  CAS  Google Scholar 

  • Bernabucci U, Lacetera N, Baumgard LH, Rhoads RP, Ronchi B, Nardone A (2010) Metabolic and hormonal adaptations to heat stress in domesticated ruminants. Animal 4:1167–1183

    Article  Google Scholar 

  • Burdon R (1986) Heat shock and the heat shock proteins. Biochem J 240:313–324

    PubMed  CAS  Google Scholar 

  • Cheng WJ, Li QL, Wang CF, Wang HM, Li JB, Sun YM, Zhong JF (2009) Genetic polymorphism of HSP70-1 gene and its correlation with resistance to mastitis in Chinese Holstein. Yi Chuan 31:169–174

    PubMed  CAS  Google Scholar 

  • Collier RJ, Stiening CM, Pollard BC, VanBaale MJ, Baumgard LH, Gentry PC, Coussens PM (2006) Use of gene expression microarrays for evaluating environmental stress tolerance at the cellular level in cattle. J Anim Sci 84(E Suppl):E1–E13

    PubMed  Google Scholar 

  • Collier RJ, Collier JL, Rhoads RP, Baumgard LH (2008) Genes involved in the bovine heat stress response. J Dairy Sci 91:445–454

    Article  PubMed  CAS  Google Scholar 

  • Duncan R (2005) Inhibition of Hsp90 function delays and impairs recovery from heat shock. FEBS J 272:5244–5256

    Article  PubMed  CAS  Google Scholar 

  • Favatier F, Bornman L, Hightower LE, Eberhand G, Polla BS (1997) Variation in Hsp gene expression and Hsp polymorphism: do they contribute to differential disease susceptibility and stress tolerance? Cell Stress Chaperones 2:141–155

    Article  PubMed  CAS  Google Scholar 

  • Grosz MD, Skow LC, Stone RT (1994) An AluI polymorphism at the bovine 70 kD heat shock protein-1 (HSP70-1) locus. Anim Genet 25:196

    Article  PubMed  CAS  Google Scholar 

  • Hansen PJ (2004) Physiological and cellular adaptations of zebu cattle to thermal stress. Anim Reprod Sci 82–83:349–360

    Article  PubMed  Google Scholar 

  • Horowitz M (2001) Heat acclimation: phenotypic plasticity and cues to the underlying molecular mechanisms. J Therm Biol 26:357–363

    Article  CAS  Google Scholar 

  • Huang L, Mivechi NF, Moskophidis D (2001) Insights into regulation and function of the major stress-induced hsp70 molecular chaperone in vivo: analysis of mice with targeted gene disruption of the hsp70.1 or hsp70.3 gene. Mol Cell Biol 21:8575–8591

    Article  PubMed  CAS  Google Scholar 

  • Huang SY, Chen MY, Lin EC, Tsou HL, Kuo YH, Ju CC, Lee WC (2002) Effects of single nucleotide polymorphisms in the 5′ flanking region of heat shock protein 70.2 gene on semen quality in boars. Anim Reprod Sci 70:99–109

    Article  PubMed  CAS  Google Scholar 

  • Hunter-Lavin C, Davies EL, Bacelar MM, Marshall MJ, Andrew SM, Williams JH (2004) Hsp 70 release from peripheral blood mononuclear cells. Biochem Biophys Res Commun 324:511–517

    Article  PubMed  CAS  Google Scholar 

  • Kampinga HH, Kanon B, Salomons FA, Kabakov AE, Patterson C (2003) Overexpression of the cochaperone CHIP enhances Hsp70-dependent folding activity in mammalian cells. Mol Cell Biol 23:4948–4958

    Article  PubMed  CAS  Google Scholar 

  • Lacetera N, Bernabucci U, Scalia D, Basiricò L, Morera P, Nardone A (2006) Heat stress elicits different responses in peripheral blood mononuclear cells from Brown Swiss and Holstein cows. J Dairy Sci 89:4606–4612

    Article  PubMed  CAS  Google Scholar 

  • Lindquist S, Craig EA (1988) The heat shock proteins. Annu Rev Genet 22:631–677

    Article  PubMed  CAS  Google Scholar 

  • Paula-Lopes FF, Chase CC Jr, Al-Katanani YM, Krininger CE, Rivera RM, Tekin S, Majewski AC, Ocon OM, Olson TA, Hansen PJ (2003) Genetic divergence in cellular resistance to heat shock in cattle: differences between breeds developed in temperate versus hot climates in responses of preimplantation embryos, reproductive tract tissues and lymphocytes to increased culture temperatures. Reproduction 125:285–294

    Article  PubMed  CAS  Google Scholar 

  • Pearce J, Thomsen S (1995) Optical-thermal response of laser-irradiated tissue. Lasers, photonics, and electro-optics. Plenum Press, New York, pp 561–603

    Google Scholar 

  • 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 15:1118–1131

    Article  PubMed  CAS  Google Scholar 

  • Rosenkrans C Jr, Banksa A, Reitera S, Looperb M (2010) Calving traits of crossbred Brahman cows are associated with heat shock protein 70 genetic polymorphisms. Anim Reprod Sci 119:178–182

    Article  PubMed  CAS  Google Scholar 

  • Schwerin M, Maak S, Kalbe C, Fuerbass R (2001) Functional promoter variants of highly conserved inducible hsp70 genes significantly affect stress response. Biochim Biophys Acta 1522:108–111

    PubMed  CAS  Google Scholar 

  • Schwerin M, Maak S, Hagendorf A, von Lengerken G, Seyfert HM (2002) A 3′-UTR variant of the inducible porcine hsp70.2 gene affects mRNA stability. Biochim Biophys Acta 1578:90–94

    PubMed  CAS  Google Scholar 

  • Singh R, Kolvraa S, Bross P, Jensen UB, Gregersen N, Tan Q, Knudsen C, Rattan SIS (2006) Reduced heat shock response in human mononuclear cells during aging and its association with polymorphisms in HSP70 genes. Cell Stress Chaperones 11:208–215

    Article  PubMed  CAS  Google Scholar 

  • Sonna LA, Gaffin SL, Pratt RE, Cullivam ML, Angel KG, Lilly CM (2002a) Effects of acute heat shock on gene expression by human peripheral blood mononuclear cells. J Appl Physiol 92:2208–2220

    PubMed  CAS  Google Scholar 

  • Sonna LA, Fujita J, Gaffin SL, Lilly CM (2002b) Invited review: effects of heat and cold stress on mammalian gene expression. J Appl Physiol 92:1725–1742

    PubMed  CAS  Google Scholar 

  • Stankiewicz AR, Lachapelle G, Foo CP, Radicioni SM, Mosser DD (2005) Hsp70 inhibits heat-induced apoptosis upstream of mitochondria by preventing Bax translocation. J Biol Chem 280:38729–38739

    Article  PubMed  CAS  Google Scholar 

  • Starkey L, Looper ML, Banks A, Reiter S, Rosenkrans C Jr (2007) Identification of polymorphisms in the promoter region of the bovine heat shock protein gene and associations with bull calf weaning weight. Am Soc Anim Sci South Sect Meet 85(suppl 2):42

    Google Scholar 

  • Steel GJ, Fullerton DM, Tyson JR, Stirling CJ (2004) Coordinated activation of Hsp70 chaperones. Science 303:98–101

    Article  PubMed  CAS  Google Scholar 

  • Stephens M, Scheet P (2005) Accounting for decay of linkage disequilibrium in haplotype inference and missing data imputation. Am J Hum Genet 76:449–462

    Article  PubMed  CAS  Google Scholar 

  • Stephens M, Smith NJ, Donnelly P (2001) A new statistical method for haplotype reconstruction from population data. Am J Hum Genet 68:978–989

    Article  PubMed  CAS  Google Scholar 

  • St-Pierre NR, Cobanov B, Schnitkey G (2003) Economic losses from heat stress by US livestock industries. J Dairy Sci 86(E-Suppl):E52–E77

    Article  Google Scholar 

  • Ulmasov KA, Shammakov S, Karaev K, Evgen'ev MB (1992) Heat shock proteins and thermoresistance in lizards. Proc Natl Acad Sci USA 89:1666–1670

    Article  PubMed  CAS  Google Scholar 

  • Zhang X, Du H, Li J (2002) Single nucleotide polymorphism of chicken heat shock protein 70 gene. 7th World Congress on Genetics Applied to Livestock Production, Montpellier, France

Download references

Acknowledgment

The research was financially supported by the SELMOL-Project (Italian Ministry of Agricultural, Food, and Forestry Policies).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Umberto Bernabucci.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Basiricò, L., Morera, P., Primi, V. et al. Cellular thermotolerance is associated with heat shock protein 70.1 genetic polymorphisms in Holstein lactating cows. Cell Stress and Chaperones 16, 441–448 (2011). https://doi.org/10.1007/s12192-011-0257-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12192-011-0257-7

Keywords

Navigation