Expression of various sarcomeric tropomyosin isoforms in equine striated muscles.
Abstract: In order to better understand the training and athletic activity of horses, we must have complete understanding of the isoform diversity of various myofibrillar protein genes like tropomyosin. Tropomyosin (TPM), a coiled-coil dimeric protein, is a component of thin filament in striated muscles. In mammals, four TPM genes (TPM1, TPM2, TPM3, and TPM4) generate a multitude of TPM isoforms via alternate splicing and/or using different promoters. Unfortunately, our knowledge of TPM isoform diversity in the horse is very limited. Hence, we undertook a comprehensive exploratory study of various TPM isoforms from horse heart and skeletal muscle. We have cloned and sequenced two sarcomeric isoforms of the gene called TPM1α and TPM1κ, one sarcomeric isoform of the and one of the gene, TPM2α and TPM3α respectively. By qRT-PCR using both relative expression and copy number, we have shown that TPM1α expression compared to TPM1κ is very high in heart. On the other hand, the expression of TPM1α is higher in skeletal muscle compared to heart. Further, the expression of TPM2α and TPM3α are higher in skeletal muscle compared to heart. Using western blot analyses with CH1 monoclonal antibody we have shown the high expression levels of sarcomeric TPM proteins in cardiac and skeletal muscle. Due to the paucity of isoform specific antibodies we cannot specifically detect the expression of TPM1κ in horse striated muscle. To the best of our knowledge this is the very first report on the characterization of sarcmeric TPMs in horse striated muscle.
Publication Date: 2017-06-26 PubMed ID: 28717602PubMed Central: PMC5498770DOI: 10.4314/ovj.v7i2.17Google Scholar: Lookup
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- Journal Article
Summary
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This research aims to understand the variety of specific protein gene isoforms, tropomyosin (TPM), in horses. The study contributes to our knowledge on how athletic activity and training in horses correspond with the presence of TPM in heart and skeletal muscles.
Objective of the Research
- The purpose of this research is to gain in-depth understanding of the tropomyosin (TPM) isoform diversity in horses. TPM, a key component of thin filament in striated muscles, plays a crucial role in muscle contraction. Recognizing the isoform diversity of TPM genes is essential to comprehend and enhance the athletic performance of horses. The availability of TPM gene isoforms in horses has been under-researched, hence motivating this study.
Methodology of the Research
- The study entailed a comprehensive exploration of TPM isoforms from horse heart and skeletal muscles. This was achieved through the cloning and sequencing of two sarcomeric isoforms of the TPM1 gene, TPM1α and TPM1κ, and one sarcomeric isoform each from TPM2 and TPM3 genes, namely TPM2α and TPM3α respectively.
- The study also used quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) to examine both the relative expression and copy number of the isolated isoforms. The technique helped confirm the prevalence of TPM1α compared to TPM1κ in the heart, and its higher expression in the skeletal muscle than in the heart. It was also used to show higher expressions of TPM2α and TPM3α in the skeletal muscle than in the heart.
- The research utilized western blot analyses with CH1 monoclonal antibody to validate the high expression levels of sarcomeric TPM proteins in cardiac and skeletal muscles.
Limitations of the Research
- An apparent limitation in the study was the inability to accurately detect the expression of TPM1κ in horse striated muscle due to the scarcity of isoform-specific antibodies.
Conclusion of the Research
- This research is a pioneering study in characterizing sarcmeric TPMs in horse striated muscle. It provides crucial insights into the expression and diversity of TPM isoforms in horses, contributing significantly to the existing literature on equine physiology and exercise science.
Cite This Article
APA
Dube S, Chionuma H, Matoq A, Alshiekh-Nasany R, Abbott L, Poiesz BJ, Dube DK.
(2017).
Expression of various sarcomeric tropomyosin isoforms in equine striated muscles.
Open Vet J, 7(2), 180-191.
https://doi.org/10.4314/ovj.v7i2.17 Publication
Researcher Affiliations
- Department of Medicine, SUNY Upstate Medical University, 750 East Adams Street, Syracuse, New York 13210, USA.
- Department of Medicine, SUNY Upstate Medical University, 750 East Adams Street, Syracuse, New York 13210, USA.
- University of Florida, College of Medicine-Jacksonville, Suite 1130, 841 Prudential Drive, Jacksonville, FL 32207, USA.
- Department of Medicine, SUNY Upstate Medical University, 750 East Adams Street, Syracuse, New York 13210, USA.
- Department of Medicine, SUNY Upstate Medical University, 750 East Adams Street, Syracuse, New York 13210, USA.
- Department of Medicine, SUNY Upstate Medical University, 750 East Adams Street, Syracuse, New York 13210, USA.
- Department of Medicine, SUNY Upstate Medical University, 750 East Adams Street, Syracuse, New York 13210, USA.
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Citations
This article has been cited 3 times.- Dube S, Abbott L, Randhawa S, Fan Y, Sanger JW, Sanger JM, Poiesz BJ, Dube DK. Identification of a novel TPM4 isoform transcript and comparison to the expression of other tropomyosin isoforms in bovine cardiac and skeletal muscles. Int J Biochem Mol Biol 2021;12(1):17-34.
- Dube DK, Dube S, Shrestha R, Abbott L, Randhawa S, Muthu V, Fan Y, Wang J, Sanger JM, Sanger JW, Poiesz BJ. Qualitative and quantitative evaluation of TPM transcripts and proteins in developing striated chicken muscles indicate TPM4α is the major sarcomeric cardiac tropomyosin from early embryonic life to adulthood. Cytoskeleton (Hoboken) 2018 Oct;75(10):437-449.
- Dube S, Abbott L, Randhawa S, Fan Y, Wang J, Sanger JM, Sanger JW, Poiesz BJ, Dube DK. Sarcomeric TPM3α in developing chicken. Cytoskeleton (Hoboken) 2018 Apr;75(4):174-182.
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