Cloning and expression of type III collagen in normal and injured tendons of horses.
Abstract: To clone the 5' end of type III collagen and describe its pattern of mRNA and protein expression in normal and healing tendons in horses. Methods: 14 healthy adult horses. Methods: The tensile region of collagenase-injured superficial digital flexor tendons was harvested at intervals from 1 to 24 weeks after injury. Total RNA was reverse-transcribed into cDNA for cloning and sequencing of type III collagen. Equine-specific nucleic acid probes were developed and used for northern blot analysis and in situ hybridization. Type III collagen protein and cyanogen bromide-cleaved collagen peptides were assessedby gel electrophresis. Results: Type III collagen mRNA expression and protein content increased immediately after injury and remained increased. Type III collagen was localized to the endotenon in normal tendon and in injured tendon at 1 week. At 8 and 24 weeks, expression became more widely distributed throughout the tendon parenchyma. Injured tendon contained 6 times more type I than type III collagen mRNA. Quantities of type III collagen protein were maximal in the first 4 weeks after injury (approx 33%) and then began to decrease. Conclusions: Type III collagen expression is increased initially in endotenon and subsequently in parenchyma of healing tendon; however, type III remains the minor collagen throughout the healing process. The role of type III collagen in tendon healing is not fully elucidated.
Publication Date: 2005-03-11 PubMed ID: 15757126DOI: 10.2460/ajvr.2005.66.266Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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The research explores the cloning and expression of type III collagen in both healthy and injured tendons of horses, noting an increase in expression post-injury and its role in the tendon healing process.
Methods and Study Subjects
- The study was conducted on 14 healthy adult horses.
- Superficial digital flexor tendons, which had been injured via collagenase injection, were harvested at different intervals ranging from 1 to 24 weeks post-injury.
- After extraction, the total RNA from the tendons was converted into cDNA—used for cloning and sequencing of type III collagen.
Additional Procedures
- Equine-specific nucleic acid probes were created and used during the northern blot analysis and in situ hybridization. These processes allow the identification, isolation, and examination of RNA fragments related to type III collagen.
- Levels of type III collagen protein and cyanogen bromide-cleaved collagen peptides were examined via gel electrophoresis, a method used for separating macromolecules such as proteins and nucleic acids.
Results
- Upon injury, there was an immediate increase in expression and content of type III collagen mRNA.
- In a normal tendon, type III collagen was found in the endotenon, a sheath of tissue. Similarly, in an injured tendon at 1 week, type III collagen was located in the endotenon.
- At 8 and 24 weeks, expression of the type III collagen was observed to be more widespread within the tendon parenchyma, which is the functional tissue in the tendon.
- The study also found that the injured tendon contained six times more type I collagen mRNA than type III collagen mRNA.
- The quantity of type III collagen protein was at its peak in the first four weeks post-injury (approximately 33%) and started to decrease thereafter.
Conclusion
- Type III collagen expression initially increases in the endotenon and then in the parenchyma of the healing tendon. Despite this, type III remains the minor collagen during the entire healing process.
- The full role of type III collagen in the tendon healing process remains unclear, suggesting a need for further research.
Cite This Article
APA
Dahlgren LA, Brower-Toland BD, Nixon AJ.
(2005).
Cloning and expression of type III collagen in normal and injured tendons of horses.
Am J Vet Res, 66(2), 266-270.
https://doi.org/10.2460/ajvr.2005.66.266 Publication
Researcher Affiliations
- Comparative Orthopaedics Laboratory, Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
MeSH Terms
- Animals
- Blotting, Northern / veterinary
- Cloning, Molecular
- Collagen Type III / genetics
- Collagen Type III / metabolism
- Gene Expression
- Horse Diseases / metabolism
- Horses / injuries
- Immunohistochemistry / veterinary
- In Situ Hybridization / veterinary
- Polymerase Chain Reaction / veterinary
- RNA, Messenger / metabolism
- Tendon Injuries / metabolism
- Tendon Injuries / veterinary
- Tendons / metabolism
- Wound Healing
Grant Funding
- AR08587 / NIAMS NIH HHS
Citations
This article has been cited 6 times.- Biasutti S, Dart A, Smith M, Blaker C, Clarke E, Jeffcott L, Little C. Spatiotemporal variations in gene expression, histology and biomechanics in an ovine model of tendinopathy.. PLoS One 2017;12(10):e0185282.
- Nemoto M, Kizaki K, Yamamoto Y, Oonuma T, Hashizume K. Tenascin-C Expression in Equine Tendon-derived Cells During Proliferation and Migration.. J Equine Sci 2013;24(2):17-24.
- Södersten F, Hultenby K, Heinegård D, Johnston C, Ekman S. Immunolocalization of collagens (I and III) and cartilage oligomeric matrix protein in the normal and injured equine superficial digital flexor tendon.. Connect Tissue Res 2013;54(1):62-9.
- Thorpe CT, Udeze CP, Birch HL, Clegg PD, Screen HR. Specialization of tendon mechanical properties results from interfascicular differences.. J R Soc Interface 2012 Nov 7;9(76):3108-17.
- Watts AE, Yeager AE, Kopyov OV, Nixon AJ. Fetal derived embryonic-like stem cells improve healing in a large animal flexor tendonitis model.. Stem Cell Res Ther 2011 Jan 27;2(1):4.
- Taylor SE, Vaughan-Thomas A, Clements DN, Pinchbeck G, Macrory LC, Smith RK, Clegg PD. Gene expression markers of tendon fibroblasts in normal and diseased tissue compared to monolayer and three dimensional culture systems.. BMC Musculoskelet Disord 2009 Feb 26;10:27.
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