Coding sequences of sarcoplasmic reticulum calcium ATPase regulatory peptides and expression of calcium regulatory genes in recurrent exertional rhabdomyolysis.
Abstract: Sarcolipin (SLN), myoregulin (MRLN), and dwarf open reading frame (DWORF) are transmembrane regulators of the sarcoplasmic reticulum calcium transporting ATPase (SERCA) that we hypothesized played a role in recurrent exertional rhabdomyolysis (RER). Objective: Compare coding sequences of SLN, MRLN, DWORF across species and between RER and control horses. Compare expression of muscle Ca2+ regulatory genes between RER and control horses. Methods: Twenty Thoroughbreds (TB), 5 Standardbreds (STD), 6 Quarter Horses (QH) with RER and 39 breed-matched controls. Methods: Sanger sequencing of SERCA regulatory genes with comparison of amino acid (AA) sequences among control, RER horses, human, mouse, and rabbit reference genomes. In RER and control gluteal muscle, quantitative real-time polymerase chain reaction of SERCA regulatory peptides, the calcium release channel (RYR1), and its accessory proteins calsequestrin (CASQ1), and calstabin (FKBP1A). Results: The SLN gene was the highest expressed horse SERCA regulatory gene with a uniquely truncated AA sequence (29 versus 31) versus other species. Coding sequences of SLN, MRLN, and DWORF were identical in RER and control horses. A sex-by-phenotype effect occurred with lower CASQ1 expression in RER males versus control males (P < .001) and RER females (P = .05) and higher FKBP1A (P = .01) expression in RER males versus control males. Conclusions: The SLN gene encodes a uniquely truncated peptide in the horse versus other species. Variants in the coding sequence of SLN, MLRN, or DWORF were not associated with RER. Males with RER have differential gene expression that could reflect adaptations to stabilize RYR1.
© 2019 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.
Publication Date: 2019-02-05 PubMed ID: 30720217PubMed Central: PMC6430904DOI: 10.1111/jvim.15425Google Scholar: Lookup
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Summary
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The study investigates the role of three transmembrane regulators, Sarcolipin (SLN), myoregulin (MRLN), and dwarf open reading frame (DWORF), in recurrent exertional rhabdomyolysis (RER). It concludes that while the coding sequences of these specific peptides are not related to RER, different gene expressions could contribute to RER developments in males.
Objective, Hypothesis, and Methodology
- This research attempted to understand if the transmembrane regulators of sarcoplasmic reticulum calcium transporting ATPase (SERCA) – Sarcolipin (SLN), myoregulin (MRLN), and dwarf open reading frame (DWORF) – play a significant role in recurrent exertional rhabdomyolysis (RER).
- The team speculated that these regulators may have an impact on the development of RER, a muscle disease affecting horses.
- The proteins SLN, MRLN, and DWORF were compared across different species and between RER-affected and healthy horses.
- The study included a diverse set of horses: Twenty Thoroughbreds, 5 Standardbreds, and 6 Quarter Horses with RER, alongside 39 breed-matched controls.
- Using Sanger sequencing, the researchers examined the SERCA regulatory genes and compared the amino acid sequences among not only the test and control group of horses but also human, mouse, and rabbit reference genomes.
Results
- The results showed that SLN was the highest expressed horse SERCA regulatory gene, and it encoded a uniquely truncated peptide sequence in horses compared to other species.
- The coding sequences of SLN, MRLN, and DWORF were found to be identical in both RER-infected and control horses.
- However, a sex-by-phenotype effect was observed. Males with RER showed lower expression of calsequestrin (CASQ1) gene as compared to male controls and RER affected females.
- Females, on the other hand, showed higher expression of calstabin (FKBP1A).
Conclusion
- On concluding the study, the researchers found that in horses, the SLN gene, a regulatory gene for muscle calcium regulation, encodes a uniquely truncated peptide compared to other species.
- The coding sequences of SLN, MRLN, or DWORF displayed no linkage with RER, contradicting the initial hypothesis held by the team.
- Males with RER, however, showed differing gene expressions which could potentially be adaptations to stabilize the RYR1, a calcium release channel.
The results thus suggest further research in understanding the genetic adaptations could provide additional insight on managing recurrent exertional rhabdomyolysis in horses.
Cite This Article
APA
Valberg SJ, Soave K, Williams ZJ, Perumbakkam S, Schott M, Finno CJ, Petersen JL, Fenger C, Autry JM, Thomas DD.
(2019).
Coding sequences of sarcoplasmic reticulum calcium ATPase regulatory peptides and expression of calcium regulatory genes in recurrent exertional rhabdomyolysis.
J Vet Intern Med, 33(2), 933-941.
https://doi.org/10.1111/jvim.15425 Publication
Researcher Affiliations
- McPhail Equine Performance Center, Department of Large Animal Clinical Sciences, Michigan State University, East Lansing, Michigan.
- McPhail Equine Performance Center, Department of Large Animal Clinical Sciences, Michigan State University, East Lansing, Michigan.
- McPhail Equine Performance Center, Department of Large Animal Clinical Sciences, Michigan State University, East Lansing, Michigan.
- McPhail Equine Performance Center, Department of Large Animal Clinical Sciences, Michigan State University, East Lansing, Michigan.
- McPhail Equine Performance Center, Department of Large Animal Clinical Sciences, Michigan State University, East Lansing, Michigan.
- Department of Population Health and Reproduction, University of California-Davis, Davis, California.
- Department of Animal Science, University of Nebraska-Lincoln, Lincoln, Nebraska.
- Equine Integrated Medicine, PLC, Lexington, Kentucky.
- Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota Medical School, Minneapolis, Minnesota.
- Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota Medical School, Minneapolis, Minnesota.
MeSH Terms
- Amino Acid Sequence
- Animals
- Case-Control Studies
- Female
- Gene Expression
- Horse Diseases / genetics
- Horses
- Humans
- Male
- Mice
- Muscle Proteins / genetics
- Muscle Proteins / metabolism
- Muscle, Skeletal
- Rabbits
- Rhabdomyolysis / genetics
- Rhabdomyolysis / veterinary
- Sarcoplasmic Reticulum Calcium-Transporting ATPases / genetics
Grant Funding
- D16Eq004 / Morris Animal Foundation
- Grayson Jockey Club Research Foundation
- R37 AG026160 / NIA NIH HHS
- R01 AR032961 / NIAMS NIH HHS
- R01 AG026160 / NIA NIH HHS
- R01 HL129814, R37 AG26160 / DDT NIH
- R01HL129814, R37AG26160 / Foundation for the National Institutes of Health
- R01 HL129814 / NHLBI NIH HHS
Conflict of Interest Statement
Dr. Valberg is 1 of the owners of the patent for the PSSM genetic test and receives sales income from its use. Her financial interest has been reviewed and managed by the University in accordance with its conflict of interest policies.
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Citations
This article has been cited 6 times.- Li Y, Zhu X, Feng P, Qiu C, Xue X, Sun X. Simple molecules make difference: short peptides play a novel role in slowing senescence. NPJ Aging 2025 Nov 26;11(1):104.
- Valberg SJ, Velez-Irizarry D, Williams ZJ, Henry ML, Iglewski H, Herrick K, Fenger C. Enriched Pathways of Calcium Regulation, Cellular/Oxidative Stress, Inflammation, and Cell Proliferation Characterize Gluteal Muscle of Standardbred Horses between Episodes of Recurrent Exertional Rhabdomyolysis. Genes (Basel) 2022 Oct 14;13(10).
- Autry JM, Svensson B, Carlson SF, Chen Z, Cornea RL, Thomas DD, Valberg SJ. Sarcoplasmic Reticulum from Horse Gluteal Muscle Is Poised for Enhanced Calcium Transport. Vet Sci 2021 Nov 23;8(12).
- Autry JM, Karim CB, Perumbakkam S, Finno CJ, McKenzie EC, Thomas DD, Valberg SJ. Sarcolipin Exhibits Abundant RNA Transcription and Minimal Protein Expression in Horse Gluteal Muscle. Vet Sci 2020 Nov 13;7(4).
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- Williams ZJ, Velez-Irizarry D, Petersen JL, Ochala J, Finno CJ, Valberg SJ. Candidate gene expression and coding sequence variants in Warmblood horses with myofibrillar myopathy. Equine Vet J 2021 Mar;53(2):306-315.
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