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Background  Heat stress adversely affects pig growth and reproduction performance by reducing feed intake, weight gain, farrowing rate, and litter size. Heat tolerance is an important characteristic in pigs, allowing them to mitigate the negative effects of heat stress on their physiological activities. Yet, genetic variation and signaling pathways associated with the biological processes of heat-tolerant pigs are currently not fully understood. This study examined differentially expressed genes and constructed gene co-expression networks on mRNAs of pigs under different heat-stress conditions using whole transcriptomic RNA-seq analyses. Semen parameters, including total sperm number per ejaculate, motility, normal morphology rate, droplets, and rejected ejaculate rate, were measured weekly on 12 boars for two time periods: thermoneutral (January to May), and heat stress (July to October). Boars were classified into heat-tolerant ( n  = 6) and heat-susceptible (n = 6) groups based on the variation of their ejaculate parameters across the two periods. RNA was isolated from the blood samples collected from the thermoneutral and heat stress periods for gene expression analysis.    Results  Under heat stress, a total of 66 differentially expressed genes (25 down-regulated, 41 up-regulated) were identified in heat-tolerant pigs compared to themselves during the thermoneutral period. A total of 1041 differentially expressed genes (282 down-regulated, 759 up-regulated) were identified in the comparison between heat-tolerant pigs and heat-susceptible pigs under heat stress. Weighted gene co-expression network analysis detected 4 and 7 modules with genes highly associated (r > 0.50,  p  < 0.05) with semen quality parameters in heat-tolerant and heat-susceptible pigs under the effects of heat stress, respectively.    Conclusion  This study utilized the sensitivity of semen to heat stress to discriminate the heat-tolerance ability of pigs. The gene expression profiles under the thermoneutral and heat stress conditions were documented in heat-tolerant and heat-susceptible boars. Findings contribute to the understanding of genes and biological mechanisms related to heat stress response in pigs and provide potential biomarkers for future investigations on the reproductive performance of pigs.

Transcriptome analysis identifies genes and co-expression networks underlying heat tolerance in pigs