Detection of functional polymorphisms in the hsp70 gene and association with cold stress response in Inner-Mongolia Sanhe cattle

Abstract

The genetic mechanisms underlying the cattle resilience to severe cold temperatures are still unknown. In this study, we observed that four blood biochemical parameters were significantly altered, i.e., blood adrenocorticotropic hormone (ACTH), triiodothyronine (T3), thyroxine (T4), and potassium (K +) after expose to − 32°C for 3 h. This was observed using 105 healthy Sanhe heifers with similar weight (398.17 ± 34.06 kg) and age (19.30 ± 4.91 months). A total of 20 single nucleotide polymorphisms (SNPs) were identified in 5′-flanking region of the hsp70 gene in Sanhe cattle, while only 10 SNPs were segregating when comparing genetic variations between Sanhe cattle and 285 Chinese Holstein samples. Statistically significant associations between the genomic markers SNP-42 − , SNP-105 + , SNP-181 + , and SNP-205 + with blood T3 and between SNP-105 + and blood T4 were observed by applying the general linear model procedure and Bonferroni t test. Furthermore, we demonstrated that the T alleles of SNP-42 − and SNP-205 + in the GC box and Kozak sequence of the hsp70 gene, respectively, significantly decreased the green fluorescent proteins activity in vitro GFP reporter assays. These findings suggest that these two SNPs are causative polymorphisms involved in the regulation of hsp70 promoter activity and might contribute to the observed association between the hsp70 gene and T3 and T4 levels in Sanhe cattle. Thus, hsp70 gene is a promising candidate gene to be validated in independent cattle populations and functional studies related to cold stress resilience in cattle.

Selection and implementation of SNP markers for parentage analysis in a Chinese crossbred cattle population

“…The selection criteria were: 1) the reported minor allele frequency (MAF) larger than 0.3, and 2) the genetic distance of markers in the same chromosome should be greater than 5 cM. Then, SNPs were re-selected according to their heterozygosity that was predicted by the direct sequencing in polymerase chain reaction (PCR) product of pooling DNA according to the previous study [36]. Only SNPs with an allele peak height ratio over 1:3 were considered to be highly heterozygosis and were reserved preferentially.…”

Selection and implementation of SNP markers for parentage analysis in a Chinese crossbred cattle population

“…The selection criteria were: 1) the reported minor allele frequency (MAF) larger than 0.3, and 2) the genetic distance of markers in the same chromosome should be greater than 5cM. Then, SNPs were re-selected according to their heterozygosity that was predicted by the direct sequencing in polymerase chain reaction (PCR) product of pooling DNA according to the previous study [19]. Only SNPs with an allele peak height ratio over 1:3 were considered to be highly heterozygosis and were reserved preferentially.…”

Association Analysis of Polymorphisms in the 5′ Flanking Region of the HSP70 Gene with Blood Biochemical Parameters of Lactating Holstein Cows under Heat and Cold Stress

“…Within-breed genetic variation exists for thermo-tolerance and disease resistance [26,27], and thus, genetic and genomic selection for improved heat or cold resistance may increase the resilience and well-being of dairy cattle [2]. Moreover, it will have important implications in the productivity, and long-term sustainability of the dairy industry as the incidence of extreme temperatures is becoming inevitable in the majority of geographic regions around the world [28,29].…”

“…HSP70 is one of the key genes from the heat shock protein (HSP) family, which are associated with thermal stress response [30][31][32][33]. The heat shock protein 70 (HSP70) is involved in many biological activities, including the proper folding of new and abnormal proteins, apoptosis, and cellular immunity [27,33]. Furthermore, HSP70 accomplishes a key role in the normal physiological activities of cells.…”

“…Polymorphisms in the HSP70 gene in different sheep breeds raised under thermal stress have shown significant influence (p < 0.05) on the blood metabolites such as glucose, serum glutamic-oxaloacetic transaminase (SGOT), phosphorous, triglycerides, and cholesterol [40]. Furthermore, a study conducted by Hu et al reported significant alteration in four blood metabolites in cattle under cold stress; ACTH (adreno-corticotropic hormone), T3 (tri-iodothyronine), T4 (thyroxine), and K+ (potassium) [27].…”