Expression of Transient Receptor Potential Vanilloid (TRPV) channels in different passages of articular chondrocytes

Ion channels play important roles in chondrocyte mechanotransduction. The transient receptor potential vanilloid (TRPV) subfamily of ion channels consists of six members. TRPV1-4 are temperature sensitive calcium-permeable, relatively non-selective cation channels whereas TRPV5 and TRPV6 show high s...

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Published in:International journal of molecular sciences 2012-04, Vol.13 (4), p.4433-4445
Main Authors: Hdud, Ismail M, El-Shafei, Abdelrafea A, Loughna, Paul, Barrett-Jolley, Richard, Mobasheri, Ali
Format: Article
Language:English
Subjects:
Animals
Biomechanics
Calcium - metabolism
Calcium Signaling - physiology
Cartilage
Cartilage, Articular - cytology
Cartilage, Articular - metabolism
Cation Transport Proteins - biosynthesis
Cell Dedifferentiation
Cell Membrane - physiology
Cells, Cultured
Chondrocytes - cytology
Chondrocytes - metabolism
Gene Expression Regulation
Horses
Load
Localization
Mechanotransduction, Cellular - physiology
Metabolism
Metabolites
Polyclonal antibodies
Signal transduction
TRPV Cation Channels - biosynthesis
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Summary:Ion channels play important roles in chondrocyte mechanotransduction. The transient receptor potential vanilloid (TRPV) subfamily of ion channels consists of six members. TRPV1-4 are temperature sensitive calcium-permeable, relatively non-selective cation channels whereas TRPV5 and TRPV6 show high selectivity for calcium over other cations. In this study we investigated the effect of time in culture and passage number on the expression of TRPV4, TRPV5 and TRPV6 in articular chondrocytes isolated from equine metacarpophalangeal joints. Polyclonal antibodies raised against TRPV4, TRPV5 and TRPV6 were used to compare the expression of these channels in lysates from first expansion chondrocytes (P0) and cells from passages 1-3 (P1, P2 and P3) by western blotting. TRPV4, TRPV5 and TRPV6 were expressed in all passages examined. Immunohistochemistry and immunofluorescence confirmed the presence of these channels in sections of formalin fixed articular cartilage and monolayer cultures of methanol fixed P2 chondrocytes. TRPV5 and TRPV6 were upregulated with time and passage in culture suggesting that a shift in the phenotype of the cells in monolayer culture alters the expression of these channels. In conclusion, several TRPV channels are likely to be involved in calcium signaling and homeostasis in chondrocytes.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms13044433