J Vet Sci.  2018 Jan;19(1):99-106. 10.4142/jvs.2018.19.1.99.

Integrated analysis of microRNA and mRNA expressions in peripheral blood leukocytes of Warmblood horses before and after exercise

• Kim HA
• Kim MC
• Kim NY
• Ryu DY
• Lee HS
• Kim Y

• KMID: 2402700
• DOI: http://doi.org/10.4142/jvs.2018.19.1.99

Abstract

Exercise capacity is a valuable trait in horses, and it has been used as a horse selection criterion. Although exercise affects molecular homeostasis and adaptation in horses, the mechanisms underlying these effects are not fully described. This study was carried out to identify changes in the blood profiles of microRNAs (miRNAs) and mRNAs induced by exercise in horse leukocytes. Total RNAs isolated from the peripheral blood leukocytes of four Warmblood horses before and after exercise were subjected to next-generation sequencing (NGS) and microarray analyses to determine the miRNA and mRNA expression profiles, respectively. The expressions of 6 miRNAs, including 4 known and 2 novel miRNAs, were altered by exercise. The predicted target genes of the differentially expressed miRNAs identified by NGS were matched to the exercise-induced mRNAs determined by microarray analysis. Five genes (LOC100050849, LOC100054517, KHDRBS3, LOC100053996, and LOC100062720) from the microarray analysis were matched to the predicted target genes of the 6 miRNAs. The subset of mRNAs and miRNAs affected by exercise in peripheral blood leukocytes may be useful in elucidating the molecular mechanisms of exercise-associated physiology in horses.

Keyword

exercise; horses; messenger RNA; microRNAs; next-generation sequencing

MeSH Terms

Homeostasis
Horses*
Leukocytes*
Microarray Analysis
MicroRNAs*
Physiology
RNA
RNA, Messenger*
MicroRNAs
RNA
RNA, Messenger

Figure

• 

  Fig. 1 Processes in the analysis of the expression of leukocyte microRNAs and mRNAs that were changed by exercise. NGS, next-generation sequencing.

• 

  Fig. 2 Characteristics of the novel microRNAs (miRNAs) affected by exercise in equine peripheral blood leukocytes. The structure of the two novel miRNAs. (A) Novel_mir_14-5p and (B) novel_mir_95-3p. The box indicates the mature sequence of the novel miRNA.

• 

  Fig. 3 Differentially expressed genes from the microarray analysis before and after exercise of three horses (No. 1, 2, and 4). (A) Significantly increased genes based on log2 (fold change) > 1 or < −1. (B) Significantly decreased genes based on log2 (fold change) > 1 or < −1. Each circle in panel A and B indicate the number of genes differentially expressed in the peripheral leukocytes of horse No. 1, 2, and 4, respectively. (C) The number of differentially expressed genes in the three horses.

• 

  Fig. 4 Regulation of microRNAs (miRNAs) and mRNAs is altered by exercise. (A) and (B) illustrate the relationships between the differentially expressed mRNAs in the microarray data and the mRNAs targeted by the differentially expressed miRNAs induced by exercise identified in the next-generation sequencing data.

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