Molecular analysis of alternative transcripts of equine AXL receptor tyrosine kinase gene.
Abstract: Since athletic performance is a most importance trait in horses, most research focused on physiological and physical studies of horse athletic abilities. In contrast, the molecular analysis as well as the regulatory pathway studies remain insufficient for evaluation and prediction of horse athletic abilities. In our previous study, we identified receptor tyrosine kinase () gene which was expressed as alternative spliced isoforms in skeletal muscle during exercise. In the present study, we validated two alternative splicing transcripts (named as for long form and for short form) in equine skeletal muscle to gain insight(s) into the role of each alternative transcript during exercise. Methods: We validated two isoforms of transcripts in horse tissues by reverse transcriptase polymerase chain reaction (RT-PCR), and then cloned the transcripts to confirm the alternative locus and its sequences. Additionally, we examined the expression patterns of and transcripts in horse tissues by quantitative RT-PCR (qRT-PCR). Results: Both of and transcripts were expressed in horse skeletal muscle and the expression levels were significantly increased after exercise. The sequencing analysis showed that there was an alternative splicing event at exon 11 between and transcripts. 3-dimentional (3D) prediction of the alternative protein structures revealed that the structural distance of the connective region between fibronectin type 3 (FN3) and immunoglobin (Ig) domain was different between two alternative isoforms. Conclusions: It is assumed that the expression patterns of and transcripts would be involved in regulation of exercise-induced stress in horse muscle possibly through an NF-κB signaling pathway. Further study is necessary to uncover biological function(s) and significance of the alternative splicing isoforms in race horse skeletal muscle.
Publication Date: 2017-08-23 PubMed ID: 28854781PubMed Central: PMC5582333DOI: 10.5713/ajas.17.0409Google Scholar: Lookup
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Summary
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The research article discusses a molecular analysis of different versions of the AXL receptor tyrosine kinase gene in horses, aiming to better understand the gene’s role in athletic performance. Two distinct transcripts of this gene were identified, their structure was analyzed, and their expression levels were measured before and after exercise.
Methodology
- The team of scientists validated the existence of two different versions of the gene’s transcript, referred to as AXL-L for the long form and AXL-S for the short form, in horse tissue. This validation process was carried out using RT-PCR, a laboratory technique used to amplify and measure specific DNA sequences.
- The transcripts were cloned to confirm alternative splicing location and sequence.
- The team also investigated how the expression patterns of these two transcripts changed in response to exercise, using quantitative RT-PCR to measure changes in expression levels.
Findings
- The results proved the existence of both AXL-L and AXL-S transcripts in horse skeletal muscle, and their expression levels increased significantly after exercise.
- An alternative splicing event was identified at exon 11 that differentiated AXL-L and AXL-S.
- The 3D prediction of the altered protein structures showed that the spacing between FN3 and the Ig domain varied between the two alternative transcripts.
Implications and Conclusions
- The researchers hypothesize that the varied expression patterns of AXL-L and AXL-S might play a role in managing exercise-induced stress in horse muscles, possibly involving an NF-κB signaling pathway. This pathway is essential to many biological processes, including inflammation and immunity, growth, and response to stress.
- The team concludes that further research is required to fully understand the biological functions and implications of the variation in these splicing isoforms in racing horse skeletal muscles. Such discoveries could potentially contribute to optimizing horse’s athletic abilities and resilience to intensive physical exercises.
Cite This Article
APA
Park JW, Song KD, Kim NY, Choi JY, Hong SA, Oh JH, Kim SW, Lee JH, Park TS, Kim JK, Kim JG, Cho BW.
(2017).
Molecular analysis of alternative transcripts of equine AXL receptor tyrosine kinase gene.
Asian-Australas J Anim Sci, 30(10), 1471-1477.
https://doi.org/10.5713/ajas.17.0409 Publication
Researcher Affiliations
- Department of Animal Science, College of Natural Resources and Life Sciences, Pusan National University, Miryang 50463, Korea.
- Department of Animal Biotechnology, Chonbuk National, University, Jeonju 54896, Korea.
- National Institute of Animal Science, Rural Development Administration, Jeju 63242, Korea.
- Department of Animal Science, College of Natural Resources and Life Sciences, Pusan National University, Miryang 50463, Korea.
- Department of Animal Science, College of Natural Resources and Life Sciences, Pusan National University, Miryang 50463, Korea.
- Department of Animal Science, College of Natural Resources and Life Sciences, Pusan National University, Miryang 50463, Korea.
- Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang 25354, Korea.
- Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang 25354, Korea.
- Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang 25354, Korea.
- Department of Microbiology, College of Natural Sciences, Changwon National University, Changwon 51140, Korea.
- Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang 25354, Korea.
- Department of Animal Science, College of Natural Resources and Life Sciences, Pusan National University, Miryang 50463, Korea.
Conflict of Interest Statement
. We certify that there is no conflict of interest with any financial organization regarding the material discussed in the manuscript.
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Citations
This article has been cited 4 times.- Park JW, Kim KH, Choi JK, Park TS, Song KD, Cho BW. Regulation of toll-like receptors expression in muscle cells by exercise-induced stress.. Anim Biosci 2021 Oct;34(10):1590-1599.
- Shen L, Lei S, Zhang B, Li S, Huang L, Czachor A, Breitzig M, Gao Y, Huang M, Mo X, Zheng Q, Sun H, Wang F. Skipping of exon 10 in Axl pre-mRNA regulated by PTBP1 mediates invasion and metastasis process of liver cancer cells.. Theranostics 2020;10(13):5719-5735.
- Lauter M, Weber A, Torka R. Targeting of the AXL receptor tyrosine kinase by small molecule inhibitor leads to AXL cell surface accumulation by impairing the ubiquitin-dependent receptor degradation.. Cell Commun Signal 2019 Jun 6;17(1):59.
- Lee HG, Choi JY, Park JW, Park TS, Song KD, Shin D, Cho BW. Effects of exercise on myokine gene expression in horse skeletal muscles.. Asian-Australas J Anim Sci 2019 Mar;32(3):350-356.
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