Enzyme-linked immunosorbent assay for myosin heavy chains in the horse.
Abstract: The content in slow and fast myosin heavy chains (MHC 1 and MHC 2) of 5 equine muscles was determined using an enzyme-linked immunosorbent assay. The results obtained with this immunoenzymatic method were compared with complementary techniques: electrophoresis and immunohistochemistry. Slices of masseter, diaphragm, tensor faciae latae, semitendinosus and cutaneus trunci were obtained from a 12-year-old saddle horse after slaughter. Muscular proteins were specifically extracted to be analysed by ELISA. The technique used 2 complimentary monoclonal antibodies (MAb). MAb 1 was prepared from a human atrium specimen that reacted specifically against MHC 1. Mab 2 was prepared from myosin of rabbit psoas muscle and reacted against MHC 2. The masseter muscle contained solely MHC 1 (100%) and this was confirmed by electrophoresis and immunohistochemistry. By contrast, the cutaneus trunci was very poor in MHC 1 (1.3%) and was entirely composed of MHC 2 (98.7%) which was confirmed by the other techniques. The diaphragm, tensor fasciae latae and semitendinosus contained 89, 40 and 2% of MHC 1, respectively. It was concluded that this ELISA method made it possible to measure a wide range of MHC contents in equine muscles with a good reproducibility. The results were consistent with those of the other fibre typing techniques. Moreover, this immunoenzymatic method is less time consuming than histological techniques and therefore offers new perspectives for muscle fibre typing in the horse.
Publication Date: 1995-01-01 PubMed ID: 8534356DOI: 10.1051/rnd:19950602Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Antibodies
- Biochemistry
- Clinical Study
- Comparative Study
- Diagnostic Technique
- Electrophoresis
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Equine Diseases
- Equine Health
- Horses
- Immunohistochemistry
- Immunology
- In Vitro Research
- Laboratory Methods
- Monoclonal Antibodies
- Muscle
- Muscle Fiber Types
- Physiology
- Veterinary Medicine
- Veterinary Research
Summary
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This research article details an examination method using the enzyme-linked immunosorbent assay (ELISA) to determine the presence of myosin heavy chains (MHC) in horse muscles. The study shows that ELISA can be effectively used to analyze and measure MHC content in equine muscles, offering a quicker alternative to traditional histological techniques.
Objectives of the Research
- The primary aim of the study was to test the efficiency and accuracy of using ELISA to measure MHC in horse muscles.
- It also aimed to compare the results acquired from ELISA with results from other techniques like electrophoresis and immunohistochemistry.
Methodology of the Research
- The researchers procured samples of five equine muscles: the masseter, diaphragm, tensor faciae latae, semitendinosus, and cutaneus trunci from a 12-year-old horse after slaughter.
- They then extracted the muscular proteins specifically for ELISA analysis.
- The ELISA technique used two complimentary monoclonal antibodies (MAb 1 and MAb 2) prepared from a human atrium specimen and rabbit psoas muscle respectively.
- MHC 1 was detected using MAb 1, while MHC 2 was identified using MAb2.
Findings of the Research
- The masseter muscle was found to contain solely MHC 1, while the cutaneus trunci contained a minuscule amount of MHC 1 and predominantly MHC 2.
- The diaphragm, tensor faciae latae, and semitendinosus contained different percentages of MHC 1 (89%, 40%, and 2%, respectively).
- All the findings from the ELISA were corroborated with results from electrophoresis and immunohistochemistry.
Conclusion of the Research
- The use of ELISA for measuring MHC in horse muscles proved to be reliable and accurate.
- The technique offers a time-saving approach compared to histological methods for muscle fiber typing in horses.
- This indicates that the ELISA method can be used widely to measure a range of MHC content in equine muscles, thus opening new avenues in the field of equine muscular research.
Cite This Article
APA
Barrey E, Valette JP, Jouglin M, Picard B, Geay Y, Robelin J.
(1995).
Enzyme-linked immunosorbent assay for myosin heavy chains in the horse.
Reprod Nutr Dev, 35(6), 619-628.
https://doi.org/10.1051/rnd:19950602 Publication
Researcher Affiliations
- INRA, station de génétique quantitative et appliquée, Jouy-en-Josas, France.
MeSH Terms
- Animals
- Electrophoresis
- Enzyme-Linked Immunosorbent Assay / methods
- Horses
- Humans
- Immunohistochemistry
- Muscle Fibers, Fast-Twitch / chemistry
- Muscle Fibers, Slow-Twitch / chemistry
- Muscles / chemistry
- Myosin Heavy Chains / analysis
- Rabbits
Citations
This article has been cited 1 times.- Rivero JL, Serrano AL, Barrey E, Valette JP, Jouglin M. Analysis of myosin heavy chains at the protein level in horse skeletal muscle.. J Muscle Res Cell Motil 1999 Feb;20(2):211-21.
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