Equine laminitis: Membrane type matrix metalloproteinase-1 (MMP-14) is involved in acute phase onset

• Published in: Equine veterinary journal 2008-07, Vol.40 (5), p.482-487
• Main Authors: Kyaw-Tanner, M. T., Wattle, O., Van Eps, A. W., Pollitt, C. C.
• Format: Article
• Language: English
• Subjects: Acute-Phase Reaction - enzymology; Acute-Phase Reaction - pathology; Acute-Phase Reaction - veterinary; Animals; DNA, Complementary - chemistry; DNA, Complementary - genetics; Foot Diseases - enzymology; Foot Diseases - pathology; Foot Diseases - veterinary; Gene Expression Regulation, Developmental; Gene Expression Regulation, Enzymologic; Hoof and Claw - enzymology; Hoof and Claw - pathology; horse; Horse Diseases - enzymology; Horse Diseases - pathology; Horses; Immunohistochemistry - veterinary; Inflammation - enzymology; Inflammation - pathology; Inflammation - veterinary; laminitis; Matrix Metalloproteinase 1 - genetics; Matrix Metalloproteinase 1 - metabolism; Matrix Metalloproteinase 14 - genetics; Matrix Metalloproteinase 14 - metabolism; matrix metalloproteinases; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - veterinary; RT-PCR TIMPs; Tissue Inhibitor of Metalloproteinase-1 - genetics; Tissue Inhibitor of Metalloproteinase-1 - metabolism; Tissue Inhibitor of Metalloproteinase-2 - genetics; Tissue Inhibitor of Metalloproteinase-2 - metabolism; Transcription, Genetic
• ISSN: 0425-1644; 2042-3306
• DOI: 10.2746/042516408X270353

Summary Reasons for performing study: Enzymatic separation at the hoof lamellar dermal‐epidermal interface may play a role in the development of laminitis and characterising and locating matrix metalloproteinases (MMPs) and their inhibitors (tissue inhibitors of MMPs or TIMPs) in lamellar tissues may further understanding of pathogenesis. Objectives: To clone and sequence the cDNA encoding lamellar MMP‐14 and TIMP‐2, and quantify their transcription in normal and laminitic tissue; and to develop antibody to locate MMP‐14 in lamellar tissues. Methods: Tissue samples were obtained from an oligofructose induced model of laminitis. Total RNA was isolated, amplified by RT‐PCR, cloned into a vector and sequenced. Real‐time PCR was used to quantify MMP‐14 and TIMP‐2 expression. Rabbit anti‐equine MMP‐14 antibody was developed to analyse MMP‐14 proteins from hoof tissues. Results: Immunohistochemistry detected MMP‐14 in the cytoplasm of normal lamellar basal and parabasal cells in close proximity to the lamellar basement membrane. In laminitis affected tissue MMP‐14 immunostaining was depleted in lamellar basal cells. Quantitative real‐time PCR showed MMP‐14 and TIMP‐2 expression significantly (P