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Validation of exercise-response genes in skeletal muscle cells of Thoroughbred racing horses

  • Kim, Doh Hoon (Department of Pathology, School of Medicine, Institute of Biomedical Science and Technology, Konkuk University) ;
  • Lee, Hyo Gun (Department of Animal Science, College of Natural Resources and Life Sciences, Pusan National University) ;
  • Sp, Nipin (Department of Pathology, School of Medicine, Institute of Biomedical Science and Technology, Konkuk University) ;
  • Kang, Dong Young (Department of Pathology, School of Medicine, Institute of Biomedical Science and Technology, Konkuk University) ;
  • Jang, Kyoung-Jin (Department of Pathology, School of Medicine, Institute of Biomedical Science and Technology, Konkuk University) ;
  • Lee, Hak Kyo (Department of Animal Biotechnology, Chonbuk National University) ;
  • Cho, Byung-Wook (Department of Animal Science, College of Natural Resources and Life Sciences, Pusan National University) ;
  • Yang, Young Mok (Department of Pathology, School of Medicine, Institute of Biomedical Science and Technology, Konkuk University)
  • Received : 2018.10.08
  • Accepted : 2019.01.28
  • Published : 2021.01.01

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 quantitative reverse transcription-polymerase chain reaction (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. The 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

References

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