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001-es BibID:BIBFORM082763
035-os BibID:(cikkazonosító)166 (WOS)000513236100002 (Scopus)85076716316
Első szerző:Matta Csaba (molekuláris biológus, genetikus, angol szakfordító)
Cím:N-methyl-D-aspartate (NMDA) receptor expression and function is required for early chondrogenesis / Csaba Matta, Tamás Juhász, János Fodor, Tibor Hajdú, Éva Katona, Csilla Szűcs-Somogyi, Roland Takács, Judit Vágó, Tamás Oláh, Ádám Bartók, Zoltan Varga, Gyorgy Panyi, László Csernoch, Róza Zákány
Megjegyzések:Background In vitro chondrogenesis depends on the concerted action of numerous signalling pathways, many of which are sensitive to the changes of intracellular Ca2+ concentration. N-methyl-D-aspartate (NMDA) glutamate receptor is a cation channel with high permeability for Ca2+. Whilst there is now accumulating evidence for the expression and function of NMDA receptors in non-neural tissues including mature cartilage and bone, the contribution of glutamate signalling to the regulation of chondrogenesis is yet to be elucidated. Methods We studied the role of glutamatergic signalling during the course of in vitro chondrogenesis in high density chondrifying cell cultures using single cell fluorescent calcium imaging, patch clamp, transient gene silencing, and western blotting. Results Here we show that key components of the glutamatergic signalling pathways are functional during in vitro chondrogenesis in a primary chicken chondrogenic model system. We also present the full glutamate receptor subunit mRNA and protein expression profile of these cultures. This is the first study to report that NMDA-mediated signalling may act as a key factor in embryonic limb bud-derived chondrogenic cultures as it evokes intracellular Ca2+ transients, which are abolished by the GluN2B subunit-specific inhibitor ifenprodil. The function of NMDARs is essential for chondrogenesis as their functional knock-down using either ifenprodil or GRIN1 siRNA temporarily blocks the differentiation of chondroprogenitor cells. Cartilage formation was fully restored with the re-expression of the GluN1 protein. Conclusions We propose a key role for NMDARs during the transition of chondroprogenitor cells to cartilage matrix-producing chondroblasts.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Glutamate signalling
N-methyl-D-aspartate receptor
Single cell calcium imaging
Megjelenés:Cell Communication and Signaling. - 17 : 1 (2019), p. 166. -
További szerzők:Juhász Tamás (1976-) (biológus, orvosbiológus) Fodor János (1973-) (élettanász, biotechnológus) Hajdú Tibor (1988-) (általános orvos) Katona Éva (1986-) (molekuláris biológus) Somogyi Csilla (1983-) (biológus, angol-magyar szakfordító) Takács Roland Ádám (1985-) (molekuláris biológus, biokémikus) Vágó Judit (1990-) (molekuláris biológus) Oláh Tamás (1983-) (élettanász) Bartók Ádám (1984-) (biotechnológus) Varga Zoltán (1969-) (biofizikus, szakfordító) Panyi György (1966-) (biofizikus) Csernoch László (1961-) (élettanász) Zákány Róza (1963-) (anatómus-, kötőszövetbiológus)
Pályázati támogatás:EFOP-3.6.2-16-2017-00006
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001-es BibID:BIBFORM028213
Első szerző:Varga Zoltán (biofizikus, szakfordító)
Cím:Switch of voltage-gated k channel expression in the plasma membrane of chondrogenic cells affects cytosolic ca-oscillations and cartilage formation / Varga Zoltán, Juhász Tamás, Matta Csaba, Fodor János, Katona Eva, Bartok Adam, Oláh Tamás, Sebe Attila, Csernoch László, Panyi György, Zákány Róza
Megjegyzések:Understanding the key elements of signaling of chondroprogenitor cells at the earliest steps of differentiation may substantially improve our opportunities for the application of mesenchymal stem cells in cartilage tissue engineering, which is a promising approach of regenerative therapy of joint diseases. Ion channels, membrane potential and Ca(2+)-signaling are important regulators of cell proliferation and differentiation. Our aim was to identify such plasma membrane ion channels involved in signaling during chondrogenesis, which may serve as specific molecular targets for influencing chondrogenic differentiation and ultimately cartilage formation. METHODOLOGY/PRINCIPAL FINDINGS:Using patch-clamp, RT-PCR and Western-blot experiments, we found that chondrogenic cells in primary micromass cell cultures obtained from embryonic chicken limb buds expressed voltage-gated Na(V)1.4, K(V)1.1, K(V)1.3 and K(V)4.1 channels, although K(V)1.3 was not detectable in the plasma membrane. Tetrodotoxin (TTX), the inhibitor of Na(V)1.4 channels, had no effect on cartilage formation. In contrast, presence of 20 mM of the K(+) channel blocker tetraethyl-ammonium (TEA) during the time-window of the final commitment of chondrogenic cells reduced K(V) currents (to 27?3% of control), cell proliferation (thymidine incorporation: to 39?4.4% of control), expression of cartilage-specific genes and consequently, cartilage formation (metachromasia: to 18.0?6.4% of control) and also depolarized the membrane potential (by 9.3?2.1 mV). High-frequency Ca(2+)-oscillations were also suppressed by 10 mM TEA (confocal microscopy: frequency to 8.5?2.6% of the control). Peak expression of TEA-sensitive K(V)1.1 in the plasma membrane overlapped with this period. Application of TEA to differentiated chondrocytes, mainly expressing the TEA-insensitive K(V)4.1 did not affect cartilage formation.CONCLUSIONS/SIGNIFICANCE:These data demonstrate that the differentiation and proliferation of chondrogenic cells depend on rapid Ca(2+)-oscillations, which are modulated by K(V)-driven membrane potential changes. K(V)1.1 function seems especially critical during the final commitment period. We show the critical role of voltage-gated cation channels in the differentiation of non-excitable cells with potential therapeutic use.
Tárgyszavak:Természettudományok Biológiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
Megjelenés:PLoS One. - 6 : 11 (2011), p. e27957. -
További szerzők:Juhász Tamás (1976-) (biológus, orvosbiológus) Matta Csaba (1980-) (molekuláris biológus, genetikus, angol szakfordító) Fodor János (1973-) (élettanász, biotechnológus) Katona Éva (1986-) (molekuláris biológus) Bartók Ádám (1984-) (biotechnológus) Oláh Tamás (1983-) (élettanász) Sebe Attila Csernoch László (1961-) (élettanász) Panyi György (1966-) (biofizikus) Zákány Róza (1963-) (anatómus-, kötőszövetbiológus)
Internet cím:Szerző által megadott URL
Intézményi repozitóriumban (DEA) tárolt változat
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