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Home / Equine Research Bank / J Vet Intern Med / Expression and activity of collagenases in the digital lamin...
Journal of veterinary internal medicine2013; 28(1); 215-222; doi: 10.1111/jvim.12252

Expression and activity of collagenases in the digital laminae of horses with carbohydrate overload-induced acute laminitis.

Abstract: Matrix metalloproteinases (MMP) are hypothesized to degrade structurally important components of the laminar extracellular matrix (ECM) in horses with laminitis. Objective: To compare levels of expression of stromelysin-1 (MMP-3), collagenases (MMP-1, -13), and membrane type-MMPs (MMP-14, -15, -16), and the distribution of their ECM substrates, in laminae of healthy horses and horses with carbohydrate overload laminitis. Methods: Twenty-five adult horses. Methods: Gene and protein expression were determined in extracts of laminae using real-time quantitative polymerase chain reaction and Western blotting after sodium dodecylsulfate polyacrylamide gel electrophoresis. Distribution of MMP-13 and ECM components was determined using indirect immunofluorescent microscopy of nonfixed frozen sections. ECM morphology was assessed by hematoxylin and eosin staining. Results: Of the genes studied, only those encoding MMP-1 and -13 were upregulated in CHO-induced laminitis; MMP-1 at Obel grade (OG)1 lameness and MMP-13 at OG3 lameness. Laminar MMP-1 was present as 52 kDa proenzyme only. MMP-13 was present as pro- (61 kDa) and processed (48 kDa) enzyme. MMP-13 localized to the basal epithelium of the secondary epidermal laminae and its increased expression were accompanied by the appearance in secondary dermal laminae (SDL) of multiple foci that were devoid of collagen I, fibronectin, chondroitin and keratan sulfate glycosaminoglycans, and eosin-staining material. Conclusions: MMP-13 is upregulated in laminae of horses with CHO-induced OG3 lameness and, by degrading components of the ECM, may contribute to the formation of ECM-free lesions (gaps or tears) that appear in the SDL with OG3 lameness.
Copyright © 2013 by the American College of Veterinary Internal Medicine.
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Publication Date: 2013-11-25 PubMed ID: 24479657PubMed Central: PMC3910380DOI: 10.1111/jvim.12252Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • N.I.H.
  • Extramural
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • Non-P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study examines how specific proteins known as matrix metalloproteinases (MMPs) behave in horses suffering from an equine foot disease called laminitis as a result of carbohydrate overload, with the findings suggesting that a certain MMP (MMP-13) may play a significant role in degrading structural components of the hooves and contributing to the disease’s progression.

Objective and Methods of the Study

  • The goal of this study was to analyze the expression levels of several types of proteins (MMPs) and their distribution within the hoof structures (ECM substrates) in healthy horses compared to horses afflicted by carbohydrate overload-induced laminitis.
  • Specific proteins of interest include stromelysin-1 (MMP-3), collagenases (MMP-1, -13), and membrane type-MMPs (MMP-14, -15, -16).
  • For the experimental procedures, the research team used gene and protein expression determination methods on extracts of hoof structures from 25 adult horses. Methods employed included real-time quantitative polymerase chain reaction and Western blotting after sodium dodecylsulfate polyacrylamide gel electrophoresis.
  • To determine the location of MMP-13 and ECM components, the researchers used indirect immunofluorescent microscopy on nonfixed frozen sections. They also analyzed ECM morphology through hematoxylin and eosin staining.

Findings of the Study

  • Of the proteins studied, only those encoding for MMP-1 and -13 showed increased activity in horses with carbohydrate-induced laminitis. This rise in activity was seen at different stages of the disease, specifically Obel grade 1 lameness for MMP-1 and Obel grade 3 for MMP-13.
  • MMP-1 was only found as a 52 kDa proenzyme. MMP-13, however, was found in both pro- (61 kDa) and processed (48 kDa) forms.
  • MMP-13 was predominantly located in the base of the secondary epidermal hoof structure. Increased MMP-13 expression was accompanied by the appearance of multiple areas within the secondary dermal hoof structures that lacked collagen I, fibronectin, chondroitin, keratan sulfate glycosaminoglycans, and eosin-staining material, which suggests degradation of ECM components.

Conclusions

  • The study concluded that MMP-13 activity is increased in the hoof structures of horses with carbohydrate-induced laminitis during Obel grade 3 lameness. This protein appears to play a significant role in degrading ECM components, potentially contributing to the formation of ECM-free lesions or defects in the hoof structure as the disease progresses.
  • This research provides critical insights on the molecular mechanisms underlying laminitis in horses and could potentially guide the development of treatments for this debilitating equine disease.

Cite This Article

APA
Wang L, Pawlak EA, Johnson PJ, Belknap JK, Alfandari D, Black SJ. (2013). Expression and activity of collagenases in the digital laminae of horses with carbohydrate overload-induced acute laminitis. J Vet Intern Med, 28(1), 215-222. https://doi.org/10.1111/jvim.12252

Publication

Journal of veterinary internal medicine
ISSN: 1939-1676
NlmUniqueID: 8708660
Country: United States
Language: English
Volume: 28
Issue: 1
Pages: 215-222

Researcher Affiliations

Wang, L
  • Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA.
Pawlak, E A
    Johnson, P J
      Belknap, J K
        Alfandari, D
          Black, S J

            MeSH Terms

            • Animals
            • Blotting, Western / veterinary
            • Extracellular Matrix / enzymology
            • Extracellular Matrix / metabolism
            • Foot Diseases / enzymology
            • Foot Diseases / metabolism
            • Foot Diseases / veterinary
            • Gene Expression Regulation, Enzymologic / physiology
            • Hoof and Claw / enzymology
            • Hoof and Claw / metabolism
            • Horse Diseases / enzymology
            • Horse Diseases / metabolism
            • Horses
            • Immunohistochemistry / veterinary
            • Matrix Metalloproteinases / genetics
            • Matrix Metalloproteinases / metabolism
            • Microscopy, Fluorescence / veterinary
            • RNA / chemistry
            • RNA / genetics
            • Real-Time Polymerase Chain Reaction / veterinary
            • Statistics, Nonparametric

            Grant Funding

            • R01 DE016289 / NIDCR NIH HHS
            • DE016289 / NIDCR NIH HHS

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            Citations

            This article has been cited 8 times.
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