Reference gene selection and myosin heavy chain (MyHC) isoform expression in muscle tissues of domestic yak (Bos grunniens)

Xiaoyun Wu; Xuelan Zhou; Xuezhi Ding; Min Chu; Chunnian Liang; Jie Pei; Lin Xiong; Pengjia Bao; Xian Guo; Ping Yan

doi:10.1371/journal.pone.0228493

Reference gene selection and myosin heavy chain (MyHC) isoform expression in muscle tissues of domestic yak (Bos grunniens)

PLoS One. 2020 Feb 6;15(2):e0228493. doi: 10.1371/journal.pone.0228493. eCollection 2020.

Authors

Xiaoyun Wu^{1

2}, Xuelan Zhou^{1

2}, Xuezhi Ding^{1

2}, Min Chu^{1

2}, Chunnian Liang^{1

2}, Jie Pei^{1

2}, Lin Xiong^{1

2}, Pengjia Bao^{1

2}, Xian Guo^{1

2}, Ping Yan^{1

2}

Affiliations

¹ Key Lab of Yak Breeding Engineering, Gansu Province, Lanzhou, China.
² Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou, China.

Abstract

Domestic yak (Bos grunniens) is the most crucial livestock in the Qinghai-Tibetan Plateau, providing meat and other necessities for local people. The skeletal muscle of adult livestock is composed of muscle fibers, and fiber composition in muscle has influence on meat qualities, such as tenderness, pH, and color. Real-time quantitative polymerase chain reaction (RT-qPCR) is a powerful tool to evaluate the gene expression of muscle fiber, but the normalization of the data depends on the stability of expressed reference genes. Unfortunately, there is no consensus for an ideal reference gene for data normalization in muscle tissues of yak. In this study, we aimed to assess the stability of 14 commonly used candidate reference genes by using five algorithms (GeNorm, NormFinder, BestKeeper, Delat Ct and Refinder). Our results suggested UXT and PRL13A were the most stable reference genes, while the most commonly used reference gene, GAPDH, was most variably expressed across different muscle tissues. We also found that the extensor digitorum lateralis (EDL), trapezius pars thoracica (TPT), and psoas major (PM) muscle had the higher content of type I muscle fibers and the lowest content of type IIB muscle fibers, while gluteobiceps (GB) muscle had the highest content of type IIB muscle fibers. Our study provides the suitable reference genes for accurate analysis of yak muscle fiber composition.

Publication types

Research Support, Non-U.S. Gov't
Validation Study

MeSH terms

Animals
Animals, Domestic / genetics
Cattle / genetics*
Gene Expression
Genes, Essential*
Male
Meat
Muscle Fibers, Skeletal / metabolism
Muscle, Skeletal / metabolism*
Myosin Heavy Chains / genetics*
Protein Isoforms / genetics
Real-Time Polymerase Chain Reaction / methods
Real-Time Polymerase Chain Reaction / standards*
Reference Standards

Substances

Protein Isoforms
Myosin Heavy Chains

Grants and funding

This study was supported by the Agricultural Science and Technology Innovation Program (CAAS-ASTIP-2014-LIHPS-01) and National Beef Cattle Industry Technology & System (CARS-37). Ping Yan received both of the awards. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.