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Asian-Australas J Anim Sci. 2017 Oct;30(10):1471-1477. doi: 10.5713/ajas.17.0409. Epub 2017 Aug 23.

Molecular analysis of alternative transcripts of equine AXL receptor tyrosine kinase gene.

Author information

1
Department of Animal Science, College of Natural Resources and Life Sciences, Pusan National University, Miryang 50463, Korea.
2
Department of Animal Biotechnology, Chonbuk National, University, Jeonju 54896, Korea.
3
National Institute of Animal Science, Rural Development Administration, Jeju 63242, Korea.
4
Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang 25354, Korea.
5
Department of Microbiology, College of Natural Sciences, Changwon National University, Changwon 51140, Korea.

Abstract

OBJECTIVE:

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 AXL receptor tyrosine kinase (AXL) gene which was expressed as alternative spliced isoforms in skeletal muscle during exercise. In the present study, we validated two AXL alternative splicing transcripts (named as AXLa for long form and AXLb 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 AXL 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 AXLa and AXLb transcripts in horse tissues by quantitative RT-PCR (qRT-PCR).

RESULTS:

Both of AXLa and AXLb 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 AXLa and AXLb 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.

CONCLUSION:

It is assumed that the expression patterns of AXLa and AXLb 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.

KEYWORDS:

AXL Receptor Tyrosine Kinase; Alternative Splicing; Athletic Performance; Horse; Muscle; RNA-Sequence

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