NMDA receptor expression and activity in osteoarthritic human articular chondrocytes

Ramage, L; Martel, M-A; Hardingham, G E; Salter, D M

Abstract

Objective: Classical neuronal signalling molecules such as substance P and glutamate have been identified in cartilage and have roles in
regulation of chondrocyte function. This study looks at expression and activity of the ionotropic glutamate NMDA (N-methyl-D-aspartic
acid) receptor (NMDAR) in human osteoarthritic (OA) chondrocytes.
Method: Chondrocytes were obtained from human knee joint arthroplasty specimens. NMDAR subunits and PSD-95 (postsynaptic density protein 95) expression were analysed by reverse transcription-polymerase chain reaction and Western blotting. Activity of NMDAR was assayed by radioactive calcium45 uptake and changes in membrane potential in the presence and absence of NMDA and NMDAR antagonists and blockade of cell membrane ion channels.
Results: NMDAR 1, 2A, 2B and PSD-95 were detected in human OA chondrocytes whereas NR2B was absent from normal chondrocytes.
NMDA induced calcium flux into OA chondrocytes and cell membrane depolarisation. These responses were blocked by NMDAR antagonists,
removal of extracellular calcium, inhibition of nNOS (neuronal nitric oxide synthase) activity and uncoupling of NMDAR from PSD-95. Blockade of sodium channels by tetrodotoxin resulted in NMDA-induced membrane hyperpolarisation which was, in turn inhibited by apamin, a blocker of SK channels. NMDA-induced changes in cell membrane potential were not affected by L-type and stretch activated calcium channel inhibitors.
Conclusions: Human OA and normal articular chondrocytes differ in the expression of NMDAR subunits. In OA chondrocytes NMDAR
signalling requires extracellular calcium, association with PSD-95, and nNOS activity. Downstream signalling results in activation of tetrodotoxin sensitive sodium channels and SK channels, a response that differs from that of normal chondrocytes suggesting altered activity of NMDAR in OA

• Journal Article Type: Article
• Acceptance Date: Apr 27, 2008
• Online Publication Date: Jun 13, 2008
• Publication Date: 2008-12
• Deposit Date: Mar 25, 2017
• Journal: Osteoarthritis and Cartilage
• Print ISSN: 1063-4584
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 16
• Issue: 12
• Pages: 1576-1584
• DOI: https://doi.org/10.1016/j.joca.2008.04.023
• Keywords: Osteoarthritis, Chondrocyte, NMDA receptor, Cell signalling,
• Public URL: http://researchrepository.napier.ac.uk/Output/819806