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Asian-Australas J Anim Sci > Accepted Articles
DOI: https://doi.org/10.5713/ajas.18.0749    [Accepted] Published online February 14, 2019.
Validation of exercise-response genes in skeletal muscle cells of Thoroughbred racing horses
Doh Hoon Kim1, Hyo Gun Lee2, Nipin Sp1, Dong Young Kang1, Kyoung-Jin Jang1, Hak Kyo Lee3, Byung-Wook Cho2,*, Young Mok Yang1,*
1Department of Pathology, School of Medicine, Institute of Biomedical Science and Technology, Konkuk University, Chungju 27478, Korea
2Department of Animal Science, College of Natural Resources and Life Sciences, Pusan National University, Miryang 50463, Korea
3Department of Animal Biotechnology, Chonbuk National University, Jeonju 54896, South Korea
Correspondence:  Byung-Wook Cho,Email: bwcho@pusan.ac.kr
Young Mok Yang, Tel: +82-10-3791-1819, Fax: +82-10-2030-7839, Email: ymyang@kku.ac.kr
Received: 8 October 2018   • Revised: 19 January 2019   • Accepted: 28 January 2019
Abstract
Objective: To understand the athletic characteristics of Thoroughbreds, high-throughput analysis has been conducted using horse muscle tissue. However, an in vitro system has been lacking for studying and validating genes from in silico data. The aim of this study is to validate genes from differentially expressed genes (DEGs) of our previous RNA-sequencing data in vitro. Also, we investigated the effects of exercise-induced stress including heat, oxidative, hypoxic and cortisol stress on horse skeletal muscle derived cells with the top six upregulated genes of DEGs.

Methods

Enriched pathway analysis was conducted using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) tool with upregulated genes in horse skeletal muscle tissue after exercise. Among the candidates, the top six genes were analysed through geneMANIA to investigate gene networks. Muscle cells derived from neonatal horse skeletal tissue were maintained and subjected to exercise-related stressors. Transcriptional changes in the top six genes followed by stressors were investigated using qRT-PCR.

Results

The inflammation response pathway was the most commonly upregulated pathway after horse exercise. Under non-cytotoxic conditions of exercise-related stressors, the transcriptional response of the top six genes was different among types of stress. Oxidative stress yielded the most similar expression pattern to DEGs.
Conclusion
Our results indicate that transcriptional change after horse exercise in skeletal muscle tissue strongly relates to stress response. qRT-PCR results showed that stressors contribute differently to the transcriptional regulation. These results would be valuable information to understand horse exercise in the stress aspect.
Keywords: Thoroughbred; Exercise Stress; Skeletal Muscle Cells; Gene Expression


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