Omneity® Pellets
All-In-One Vitamin & Mineral Pellet
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 The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
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.
References
This article includes 26 references
- Gu J, Orr N, Park SD, Katz LM, Sulimova G, MacHugh DE, Hill EW. A genome scan for positive selection in thoroughbred horses.. PLoS One 2009 Jun 2;4(6):e5767.
- Park KD, Park J, Ko J, Kim BC, Kim HS, Ahn K, Do KT, Choi H, Kim HM, Song S, Lee S, Jho S, Kong HS, Yang YM, Jhun BH, Kim C, Kim TH, Hwang S, Bhak J, Lee HK, Cho BW. Whole transcriptome analyses of six thoroughbred horses before and after exercise using RNA-Seq.. BMC Genomics 2012 Sep 12;13:473.
- O'Bryan JP, Frye RA, Cogswell PC, Neubauer A, Kitch B, Prokop C, Espinosa R 3rd, Le Beau MM, Earp HS, Liu ET. axl, a transforming gene isolated from primary human myeloid leukemia cells, encodes a novel receptor tyrosine kinase.. Mol Cell Biol 1991 Oct;11(10):5016-31.
- Sainaghi PP, Castello L, Bergamasco L, Galletti M, Bellosta P, Avanzi GC. Gas6 induces proliferation in prostate carcinoma cell lines expressing the Axl receptor.. J Cell Physiol 2005 Jul;204(1):36-44.
- McCloskey P, Fridell YW, Attar E, Villa J, Jin Y, Varnum B, Liu ET. GAS6 mediates adhesion of cells expressing the receptor tyrosine kinase Axl.. J Biol Chem 1997 Sep 12;272(37):23285-91.
- Tai KY, Shieh YS, Lee CS, Shiah SG, Wu CW. Axl promotes cell invasion by inducing MMP-9 activity through activation of NF-kappaB and Brg-1.. Oncogene 2008 Jul 3;27(29):4044-55.
- Cunningham BA, Hemperly JJ, Murray BA, Prediger EA, Brackenbury R, Edelman GM. Neural cell adhesion molecule: structure, immunoglobulin-like domains, cell surface modulation, and alternative RNA splicing.. Science 1987 May 15;236(4803):799-806.
- Fischer EH, Charbonneau H, Tonks NK. Protein tyrosine phosphatases: a diverse family of intracellular and transmembrane enzymes.. Science 1991 Jul 26;253(5018):401-6.
- Varnum BC, Young C, Elliott G, Garcia A, Bartley TD, Fridell YW, Hunt RW, Trail G, Clogston C, Toso RJ. Axl receptor tyrosine kinase stimulated by the vitamin K-dependent protein encoded by growth-arrest-specific gene 6.. Nature 1995 Feb 16;373(6515):623-6.
- Stitt TN, Conn G, Gore M, Lai C, Bruno J, Radziejewski C, Mattsson K, Fisher J, Gies DR, Jones PF. The anticoagulation factor protein S and its relative, Gas6, are ligands for the Tyro 3/Axl family of receptor tyrosine kinases.. Cell 1995 Feb 24;80(4):661-70.
- Sasaki T, Knyazev PG, Clout NJ, Cheburkin Y, Göhring W, Ullrich A, Timpl R, Hohenester E. Structural basis for Gas6-Axl signalling.. EMBO J 2006 Jan 11;25(1):80-7.
- Goruppi S, Ruaro E, Varnum B, Schneider C. Requirement of phosphatidylinositol 3-kinase-dependent pathway and Src for Gas6-Axl mitogenic and survival activities in NIH 3T3 fibroblasts.. Mol Cell Biol 1997 Aug;17(8):4442-53.
- Hasanbasic I, Cuerquis J, Varnum B, Blostein MD. Intracellular signaling pathways involved in Gas6-Axl-mediated survival of endothelial cells.. Am J Physiol Heart Circ Physiol 2004 Sep;287(3):H1207-13.
- Paccez JD, Vogelsang M, Parker MI, Zerbini LF. The receptor tyrosine kinase Axl in cancer: biological functions and therapeutic implications.. Int J Cancer 2014 Mar 1;134(5):1024-33.
- Rankin EB, Fuh KC, Castellini L, Viswanathan K, Finger EC, Diep AN, LaGory EL, Kariolis MS, Chan A, Lindgren D, Axelson H, Miao YR, Krieg AJ, Giaccia AJ. Direct regulation of GAS6/AXL signaling by HIF promotes renal metastasis through SRC and MET.. Proc Natl Acad Sci U S A 2014 Sep 16;111(37):13373-8.
- Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.. Methods 2001 Dec;25(4):402-8.
- Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees.. Mol Biol Evol 1987 Jul;4(4):406-25.
- Cho HW, Shin S, Park JW, Choi JY, Kim NY, Lee WK, Lee HK, Song KD, Cho BW. Molecular Characterization and Expression Analysis of the Peroxisome Proliferator Activated Receptor Delta (PPARδ) Gene before and after Exercise in Horse.. Asian-Australas J Anim Sci 2015 May;28(5):697-702.
- Källberg M, Wang H, Wang S, Peng J, Wang Z, Lu H, Xu J. Template-based protein structure modeling using the RaptorX web server.. Nat Protoc 2012 Jul 19;7(8):1511-22.
- Madhusudhan MS, Marti-Renom MA, Eswar N. Comparative protein structure modeling. Totowa, NJ: Humana Press; 2005; pp. 831–60.
- Rose PW, Beran B, Bi C, Bluhm WF, Dimitropoulos D, Goodsell DS, Prlic A, Quesada M, Quinn GB, Westbrook JD, Young J, Yukich B, Zardecki C, Berman HM, Bourne PE. The RCSB Protein Data Bank: redesigned web site and web services.. Nucleic Acids Res 2011 Jan;39(Database issue):D392-401.
- MacKerell AD Jr, Banavali N, Foloppe N. Development and current status of the CHARMM force field for nucleic acids.. Biopolymers 2000-2001;56(4):257-65.
- Holland SJ, Powell MJ, Franci C, Chan EW, Friera AM, Atchison RE, McLaughlin J, Swift SE, Pali ES, Yam G, Wong S, Lasaga J, Shen MR, Yu S, Xu W, Hitoshi Y, Bogenberger J, Nör JE, Payan DG, Lorens JB. Multiple roles for the receptor tyrosine kinase axl in tumor formation.. Cancer Res 2005 Oct 15;65(20):9294-303.
- Hasanbasic I, Cuerquis J, Varnum B, Blostein MD. Intracellular signaling pathways involved in Gas6-Axl-mediated survival of endothelial cells.. Am J Physiol Heart Circ Physiol 2004 Sep;287(3):H1207-13.
- Ji LL, Gomez-Cabrera MC, Steinhafel N, Vina J. Acute exercise activates nuclear factor (NF)-kappaB signaling pathway in rat skeletal muscle.. FASEB J 2004 Oct;18(13):1499-506.
- Llinás M. Metal-polypeptide interactions: The conformational state of iron proteins. Berlin, Heidelberg: Springer; 1973; pp. 135–220.
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.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
