Molecular cloning and functional expression of the K+ channel KV7.1 and the regulatory subunit KCNE1 from equine myocardium.
Abstract: The voltage-gated K-channel K7.1 and the subunit KCNE1, encoded by the KCNQ1 and KCNE1 genes, respectively, are responsible for termination of the cardiac action potential. In humans, mutations in these genes can predispose patients to arrhythmias and sudden cardiac death (SCD). Objective: To characterize equine K7.1/KCNE1 currents and compare them to human K7.1/KCNE1 currents to determine whether K7.1/KCNE1 plays a similar role in equine and human hearts. Methods: mRNA encoding K7.1 and KCNE1 was isolated from equine hearts, sequenced, and cloned into expression vectors. The channel subunits were heterologously expressed in Xenopus laevis oocytes or CHO-K1 cells and characterized using voltage-clamp techniques. Results: Equine K7.1/KCNE1 expressed in CHO-K1 cells exhibited electrophysiological properties that are overall similar to the human orthologs; however, a slower deactivation was found which could result in more open channels at fast rates. Conclusions: The results suggest that the equine K7.1/KCNE1 channel may be important for cardiac repolarization and this could indicate that horses are susceptible to SCD caused by mutations in KCNQ1 and KCNE1.
Copyright © 2017 Elsevier Ltd. All rights reserved.
Publication Date: 2017-09-11 PubMed ID: 28917093DOI: 10.1016/j.rvsc.2017.09.010Google Scholar: Lookup
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- Journal Article
Summary
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The research investigates the function of the K+ channel KV7.1 and the regulating subunit KCNE1 in horse heart tissue, assessing their relevance to cardiac functioning and their potential role in conditions like cardiac arrhythmias and sudden cardiac death.
Introduction and Objective of the Research
- The study mainly focuses on studying the K7.1 and KCNE1 currents, crucial for cardiac action potential termination in horses, and comparing them with human counterparts. This is an attempt to comprehend similar potential effects of mutations in these genes in horses, as observed in humans leading to conditions like arrhythmia and sudden cardiac death (SCD).
Methodology Employed
- Researchers isolated mRNA encoding K7.1 and KCNE1 from horse hearts, sequenced them, and cloned them into expression vectors.
- Then, the researchers expressed the K7.1/KCNE1 channel subunits heterologously in the oocytes of ‘Xenopus laevis’ (a type of frog) or in CHO-K1 cells.
- They characterized the channels using voltage-clamp techniques, a common method used for studying ion currents in cells.
Results Gathered
- The study found that the horse K7.1/KCNE1 displayed electrophysiological properties similar to the human versions when expressed in CHO-K1 cells.
- However, a slower deactivation was observed. This could result in more open channels at high frequencies which could change the potential of the cell more frequently.
Conclusion
- The study concluded that the K7.1/KCNE1 channel could play a significant role in cardiac repolarization in horses.
- Moreover, it indicated that horses might be susceptible to SCD due to mutations in KCNQ1 and KCNE1, mirroring the effects observed in humans.
Cite This Article
APA
Pedersen PJ, Thomsen KB, Flak JB, Tejada MA, Hauser F, Trachsel D, Buhl R, Kalbfleisch T, DePriest MS, MacLeod JN, Calloe K, Klaerke DA.
(2017).
Molecular cloning and functional expression of the K+ channel KV7.1 and the regulatory subunit KCNE1 from equine myocardium.
Res Vet Sci, 113, 79-86.
https://doi.org/10.1016/j.rvsc.2017.09.010 Publication
Researcher Affiliations
- Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.
- Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.
- Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.
- Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.
- Center for Functional and Comparative Insect Genomics, Department of Biology, University of Copenhagen, Copenhagen, Denmark.
- Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.
- Department of Veterinary Clinical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Taastrup, Denmark.
- Department of Biochemistry and Molecular Genetics, School of Medicine, University of Louisville, Louisville, KY, United States.
- Department of Biochemistry and Molecular Genetics, School of Medicine, University of Louisville, Louisville, KY, United States.
- Maxwell H., Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington, KY, United States.
- Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark. Electronic address: Kirstinec@sund.ku.dk.
- Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.
MeSH Terms
- Animals
- CHO Cells
- Cloning, Molecular
- Cricetulus
- Gene Expression
- Horses / metabolism
- Humans
- KCNQ1 Potassium Channel / genetics
- KCNQ1 Potassium Channel / metabolism
- Myocardium / metabolism
- Oocytes
- Potassium Channels, Voltage-Gated / genetics
- Potassium Channels, Voltage-Gated / metabolism
- Sequence Analysis, DNA / veterinary
- Xenopus laevis
Citations
This article has been cited 5 times.- Pan J, Purev C, Zhao H, Zhang Z, Wang F, Wendoule N, Qi G, Liu Y, Zhou H. Discovery of exercise-related genes and pathway analysis based on comparative genomes of Mongolian originated Abaga and Wushen horse. Open Life Sci 2022;17(1):1269-1281.
- Trachsel DS, Stage HJ, Rausch S, Trappe S, Söllig K, Sponder G, Merle R, Aschenbach JR, Gehlen H. Comparison of Sources and Methods for the Isolation of Equine Adipose Tissue-Derived Stromal/Stem Cells and Preliminary Results on Their Reaction to Incubation with 5-Azacytidine. Animals (Basel) 2022 Aug 11;12(16).
- Premont A, Saadeh K, Edling C, Lewis R, Marr CM, Jeevaratnam K. Cardiac ion channel expression in the equine model - In-silico prediction utilising RNA sequencing data from mixed tissue samples. Physiol Rep 2022 Jul;10(14):e15273.
- Avison A, Physick-Sheard PW, Pyle WG. Performance horses as a model for exercise-associated cardiac arrhythmias and sudden cardiac death. J Mol Cell Cardiol Plus 2025 Jun;12:100452.
- Kapusniak A, Nath L, Hebart M, Franklin S. Heritability of sudden cardiac death in Thoroughbred racehorses. Equine Vet J 2025 Mar;57(2):325-332.
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