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Validation of Suitable Reference Genes for Gene Expression Studies on Yak Testis Development.

Xuelan Zhou1, Xiaoyun Wu1, Min Chu1

  • 1Key Laboratory of Yak Breeding Engineering, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China.

Animals : an Open Access Journal From MDPI
|January 25, 2020
PubMed
Summary

This study identified the most stable reference genes for yak testis development using RT-qPCR. TATA box-binding protein (TBP) and ubiquitously expressed transcript protein (UXT) are recommended for accurate gene expression analysis in yaks.

Keywords:
RNAexpression stabilitynormalizationquantitative real-time polymerase chain reaction

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Area of Science:

  • Reproductive Biology
  • Molecular Genetics
  • Animal Science

Background:

  • Yak testis development is crucial for male reproduction and is governed by numerous genes.
  • Accurate gene expression analysis using real-time reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) relies on stable reference genes.
  • Identifying ideal reference genes for yak testis development is essential to avoid misinterpreting gene expression data.

Purpose of the Study:

  • To identify the most stable reference genes for normalizing gene expression data in yak testis across different developmental stages.
  • To provide reliable reference genes for future studies on yak reproductive molecular mechanisms.

Main Methods:

  • Selected 13 commonly used reference genes for analysis.
  • Evaluated gene stability using GeNorm, NormFinder, BestKeeper, and ∆Ct methods.
  • Utilized RefFinder to integrate results and determine the top-ranking reference genes.

Main Results:

  • TATA box-binding protein (TBP) and ubiquitously expressed transcript protein (UXT) demonstrated the highest stability across all developmental stages.
  • TBP and hydroxymethylbilane synthase (HMBS) were stable in immature yak testes.
  • Mitochondrial ribosomal protein L39 (MRPL39) and TBP showed high stability in mature yak testes.

Conclusions:

  • TBP and UXT are recommended as the most reliable reference genes for yak testis gene expression studies.
  • Specific reference genes (TBP/HMBS for immature, MRPL39/TBP for mature stages) can be utilized for stage-specific analyses.
  • This research provides a foundation for accurate molecular mechanism investigations in yak testis development.