Selection of species specific panel of reference genes in peripheral blood mononuclear cells of native livestock species adapted to trans-Himalayan region of Leh-Ladakh.
Abstract: The identification of appropriate references genes is an integral component of any gene expression-based study for getting accuracy and reliability in data interpretation. In this study, we evaluated the expression stability of 10 candidate reference genes (GAPDH, RPL4, EEF1A1, RPS9, HPRT1, UXT, RPS23, B2M, RPS15, ACTB) in peripheral blood mononuclear cells of livestock species that are adapted to high altitude hypoxia conditions of Leh-Ladakh. A total of 37 PBMCs samples from six native livestock species of Leh-Ladakh region such as Ladakhi cattle, Ladakhi yak, Ladakhi donkey, Chanthangi goat, Double hump cattle and Zanskar ponies were included in this study. The commonly used statistical algorithms such as geNorm, Normfinder, BestKeeper and RefFinder were employed to assess the stability of these RGs in all the livestock species. Our study has identified different panel of reference genes in each species; for example, EEF1A1, RPL4 in Ladakhi cattle; GAPDH, RPS9, ACTB in Ladakhi yak; HPRT1, B2M, ACTB in Ladakhi donkey; HPRT1, B2M, ACTB in Double hump camel, RPS9, HPRT1 in Changthangi goat, HPRT1 and ACTB in Zanskar ponies. To the best of our knowledge, this is the first systematic attempt to identify panel of RGs across different livestock species types adapted to high altitude hypoxia conditions. In future, the findings of the present study would be quite helpful in conducting any transcriptional studies to understand the molecular basis of high altitude adaptation of native livestock population of Leh-Ladakh.
© 2022. The Author(s).
Publication Date: 2022-11-02 PubMed ID: 36323741PubMed Central: PMC9630269DOI: 10.1038/s41598-022-22588-0Google Scholar: Lookup
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Summary
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This research studied the stability of several candidate reference genes in the blood cells of livestock adapted to the high-altitude conditions in Leh-Ladakh. The findings revealed different reference genes in each species, impacting future studies on high-altitude adaptation.
Objective of the Research
- The main objective of this study was to evaluate the stability of 10 candidate reference genes in peripheral blood mononuclear cells of native livestock species in Leh-Ladakh, a high-altitude region. This was done with the aim to enhance the accuracy and reliability of future gene expression-based studies.
Methods Used in the Research
- The reference genes evaluated included GAPDH, RPL4, EEF1A1, RPS9, HPRT1, UXT, RPS23, B2M, RPS15, and ACTB.
- The study looked at 37 PBMC samples from six livestock species that have adapted to the high altitude hypoxia conditions of Leh-Ladakh. These species include Ladakhi cattle, Ladakhi yak, Ladakhi donkey, Chanthangi goat, Double hump cattle, and Zanskar ponies.
- Several statistical algorithms, such as geNorm, Normfinder, BestKeeper, and RefFinder, were used to assess the stability of these reference genes across the livestock species.
Key Findings of the Research
- The results of the study identified different panels of reference genes in each species. For instance, in Ladakhi cattle, the identified reference genes were EEF1A1 and RPL4, while in Ladakhi yak, the genes were GAPDH, RPS9, and ACTB.
- This is the first systematic attempt to identify a panel of reference genes across different livestock species adapted to high-altitude hypoxia conditions.
Implications of the Research
- The findings of this research will be useful for future transcriptional studies to understand the molecular basis of high-altitude adaptation of native livestock in Leh-Ladakh.
- By identifying a specific panel of reference genes in each livestock species, this study provides a critical base for accurate gene expression studies in high-altitude livestock populations.
Cite This Article
APA
Tiwari M, Sodhi M, Verma P, Vivek P, Kataria RS, Niranjan SK, Bharti VK, Masharing N, Gujar G, Chanda D, Mukesh M.
(2022).
Selection of species specific panel of reference genes in peripheral blood mononuclear cells of native livestock species adapted to trans-Himalayan region of Leh-Ladakh.
Sci Rep, 12(1), 18473.
https://doi.org/10.1038/s41598-022-22588-0 Publication
Researcher Affiliations
- Animal Biotechnology Center, ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana, India.
- Animal Biotechnology Center, ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana, India.
- Animal Biotechnology Center, ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana, India.
- Animal Biotechnology Center, ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana, India.
- Animal Biotechnology Center, ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana, India.
- Animal Biotechnology Center, ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana, India.
- DRDO-Defense Institute of High-Altitude Research, Leh, India.
- Animal Biotechnology Center, ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana, India.
- Animal Biotechnology Center, ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana, India.
- Animal Biotechnology Center, ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana, India.
- Animal Biotechnology Center, ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana, India. mmukesh_26@hotmail.com.
MeSH Terms
- Cattle / genetics
- Horses / genetics
- Animals
- Leukocytes, Mononuclear
- Altitude Sickness
- Livestock / genetics
- Real-Time Polymerase Chain Reaction
- Reproducibility of Results
- Hypoxia / genetics
- Goats / genetics
- Equidae / genetics
- Gene Expression Profiling
- Reference Standards
Conflict of Interest Statement
The authors declare no competing interests.
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Citations
This article has been cited 2 times.- Ma J, Song N, Huang N, Rong Z, Li J, Wang S, Zhang X, Wei Q, Chen J. Effective energy harvesting of Yak by regulating the glucose and lipid metabolism in liver to adapt to the plateau environment. Sci Rep 2025 Dec 4;16(1):944.
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