Equine laminitis: increased transcription of matrix metalloproteinase-2 (MMP-2) occurs during the developmental phase


Autoria(s): Kyaw-Tanner, M.; Pollitt, C. C.
Contribuinte(s)

Peter D. Rossdale

Data(s)

01/04/2004

Resumo

Reasons for performing study: The dysadhesion and destruction of lamellar basement membrane of laminitis may be due to increased lamellar metalloproteinase activity. Characterising lamellar metalloproteinase-2 (MMP-2) and locating it in lamellar tissues may help determine if laminitis pathology is associated with increased MMP-2 transcription. Objectives: To clone and sequence the cDNA encoding lamellar MMP-2, develop antibody and in situ hybridisation probes to locate lamellar MMP-2 and quantitate MMP-2 transcription in normal and laminitis tissue. Methods: Total RNA was isolated, fragmented by RT-PCR, cloned into vector and sequenced. Rabbit anti-equine MMP-2 and labelled MMP-2 riboprobe were developed to analyse and quantitate MMP-2 expression. Results: Western immunoblotting with anti-MMP-2 detected 72 kDa MMP-2 in hoof tissue homogenates and cross-reacted with human MMP-2. Immunohistochemistry and in situ hybridisation detected MMP-2 in the cytoplasm of basal and parabasal cells in close proximity to the lamellar basement membrane. Northern analysis and quantitative real-time PCR showed MMP-2 expression significantly (P

Identificador

http://espace.library.uq.edu.au/view/UQ:71176

Idioma(s)

eng

Publicador

Equine Veterinary Journal Ltd

Palavras-Chave #Laminitis #Matrix Metalloproteinase-2 #Veterinary Sciences #Horse #Equine Laminitis #Matrix Metalloproteinases #Real-time Pcr #Cloning #Sequencing #Tissue Inhibitors #Cell-migration #Expression #Collagenase #Activation #C1 #300501 Veterinary Medicine #780105 Biological sciences
Tipo

Journal Article