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Home / Equine Research Bank / Am J Vet Res / Distribution and processing of a disintegrin and metalloprot...
American journal of veterinary research2012; 73(7); 1035-1046; doi: 10.2460/ajvr.73.7.1035

Distribution and processing of a disintegrin and metalloproteinase with thrombospondin motifs-4, aggrecan, versican, and hyaluronan in equine digital laminae.

Abstract: To determine the expression and distribution of a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), its substrates aggrecan and versican, and their binding partner hyaluronan in laminae of healthy horses. Methods: Laminae from the forelimb hooves of 8 healthy horses. Methods: Real-time quantitative PCR assay was used for gene expression analysis. Hyaluronidase, chondroitinase, and keratanase digestion of lamina extracts combined with SDS-PAGE and western blotting were used for protein and proteoglycan analysis. Immunofluorescent and immunohistochemical staining of tissue sections were used for protein and hyaluronan localization. Results: Genes encoding ADAMTS-4, aggrecan, versican, and hyaluronan synthase II were expressed in laminae. The ADAMTS-4 was predominantly evident as a 51-kDa protein bearing a catalytic site neoepitope indicative of active enzyme and in situ activity, which was confirmed by the presence of aggrecan and versican fragments bearing ADAMTS-4 cleavage neoepitopes in laminar protein extracts. Aggrecan, versican, and hyaluronan were localized to basal epithelial cells within the secondary epidermal laminae. The ADAMTS-4 localized to these cells but was also present in some cells in the dermal laminae. Conclusions: Within digital laminae, versican exclusively and aggrecan primarily localized within basal epithelial cells and both were constitutively cleaved by ADAMTS-4, which therefore contributed to their turnover. On the basis of known properties of these proteoglycans, it is possible that they can protect the basal epithelial cells of horses from biomechanical and concussive stress.
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Publication Date: 2012-06-29 PubMed ID: 22738056PubMed Central: PMC3535468DOI: 10.2460/ajvr.73.7.1035Google 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 research explores the expression and distribution of certain peptides and proteins – ADAMTS-4, aggrecan, versican, and hyaluronan – in the digital laminae of healthy horses. The study indicates that these substances, particularly ADAMTS-4, are active within these cells and could potentially offer protection against stress.

Research Background and Methods

  • The study focused on understanding the presence and distribution of the peptides ADAMTS-4, aggrecan, versican, and their binding partner hyaluronan in the digital laminae of healthy horses.
  • The research materials were sourced from the forelimb hooves of eight horses considered healthy.
  • To detect and quantify the expression of these substances, a real-time quantitative PCR assay was used, which is a test that determines the copy number of a particular DNA or RNA sequence in a sample.
  • A combination of hyaluronidase, chondroitinase, and keratanase digestion of lamina extracts, along with SDS-PAGE and western blotting procedures were used to analyze the protein and proteoglycans.
  • Furthermore, to localize the proteins and hyaluronan in tissue sections, immunofluorescent and immunohistochemical staining techniques were applied.

Research Findings

  • The research results showed that genes encoding ADAMTS-4, aggrecan, versican, and hyaluronan synthase II were found in laminae.
  • ADAMTS-4 was mainly seen as a 51-kDa protein bearing a catalytic site neoepitope, an indicator of an active enzyme, and its in situ activity was confirmed by the presence of aggrecan and versican fragments bearing ADAMTS-4 cleavage neoepitopes in laminar protein extracts.
  • The proteins aggrecan, versican, and hyaluronan were localized to basal epithelial cells within the secondary epidermal laminae.
  • ADAMTS-4 was present in these cells but was also found in some cells in the dermal laminae.

Conclusions and Implications

  • The study concluded that within digital laminae, versican exclusively and aggrecan principally localize within basal epithelial cells. Both are perpetually cleaved by ADAMTS-4, which contributes to their turnover.
  • Based on known properties of these proteoglycans, researchers hypothesized that versican and aggrecan may help protect the basal epithelial cells of horses from mechanical and concussive stress.
  • This research has implications for understanding the animal’s responses toward stress at the cellular level and might provide insights into treating related conditions in horses.

Cite This Article

APA
Pawlak E, Wang L, Johnson PJ, Nuovo G, Taye A, Belknap JK, Alfandari D, Black SJ. (2012). Distribution and processing of a disintegrin and metalloproteinase with thrombospondin motifs-4, aggrecan, versican, and hyaluronan in equine digital laminae. Am J Vet Res, 73(7), 1035-1046. https://doi.org/10.2460/ajvr.73.7.1035

Publication

American journal of veterinary research
ISSN: 1943-5681
NlmUniqueID: 0375011
Country: United States
Language: English
Volume: 73
Issue: 7
Pages: 1035-1046

Researcher Affiliations

Pawlak, Erica
  • Department of Veterinary and Animal Sciences, College of Natural Sciences, University of Massachusetts, Amherst, MA 01003, USA.
Wang, Le
    Johnson, Philip J
      Nuovo, Gerard
        Taye, Almaz
          Belknap, James K
            Alfandari, Dominique
              Black, Samuel J

                MeSH Terms

                • ADAM Proteins / genetics
                • ADAM Proteins / metabolism
                • ADAMTS4 Protein
                • Aggrecans / genetics
                • Aggrecans / metabolism
                • Animals
                • Blotting, Western / veterinary
                • Gene Expression Profiling / veterinary
                • Hoof and Claw / metabolism
                • Horses / metabolism
                • Hyaluronic Acid / genetics
                • Hyaluronic Acid / metabolism
                • Immunohistochemistry / veterinary
                • Procollagen N-Endopeptidase / genetics
                • Procollagen N-Endopeptidase / metabolism
                • RNA / chemistry
                • RNA / genetics
                • Reverse Transcriptase Polymerase Chain Reaction / veterinary
                • Versicans / genetics
                • Versicans / metabolism

                Grant Funding

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

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                Citations

                This article has been cited 5 times.
                1. James S, Daffy J, Cook J, Samiric T. Short-Term Exposure to Ciprofloxacin Reduces Proteoglycan Loss in Tendon Explants.. Genes (Basel) 2022 Nov 25;13(12).
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                5. Wang L, Pawlak E, Johnson PJ, Belknap JK, Alfandari D, Black SJ. Effects of cleavage by a disintegrin and metalloproteinase with thrombospondin motifs-4 on gene expression and protein content of versican and aggrecan in the digital laminae of horses with starch gruel-induced laminitis.. Am J Vet Res 2012 Jul;73(7):1047-56.
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