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001-es BibID:BIBFORM054229
035-os BibID:(PMID)25112418 (WOS)000344875700028 (Scopus)84939883199
Első szerző:Juhász Tamás (biológus, orvosbiológus)
Cím:Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) Signalling Enhances Osteogenesis in UMR-106 Cell Line / Tamás Juhász, Csaba Matta, Éva Katona, Csilla Somogyi, Roland Takács, Tibor Hajdú, Solveig Lind Helgadottir, János Fodor, László Csernoch, Gábor Tóth, Éva Bakó, Dóra Reglődi, Andrea Tamás, Róza Zákány
Dátum:2014
ISSN:0895-8696
Megjegyzések:Presence of the pituitary adenylate cyclase-activating polypeptide (PACAP) signalling has been proved in various peripheral tissues. PACAP can activate protein kinase A (PKA) signalling via binding to pituitary adenylate cyclase-activating polypeptide type I receptor (PAC1), vasoactive intestinal polypeptide receptor (VPAC) 1 or VPAC2 receptor. Since little is known about the role of this regulatory mechanism in bone formation, we aimed to investigate the effect of PACAP on osteogenesis of UMR-106 cells. PACAP 1-38 as an agonist and PACAP 6-38 as an antagonist of PAC1 were added to the culture medium. Surprisingly, both substances enhanced protein expressions of collagen type I, osterix and alkaline phosphatase, along with higher cell proliferation rate and an augmented mineralisation. Although expression of PKA was elevated, no alterations were detected in the expression, phosphorylation and nuclear presence of CREB, but increased nuclear appearance of Runx2, the key transcription factor of osteoblast differentiation, was shown. Both PACAPs increased the expressions of bone morphogenetic proteins (BMPs) 2, 4, 6, 7 and Smad1 proteins, as well as that of Sonic hedgehog, PATCH1 and Gli1. Data of our experiments indicate that activation of PACAP pathway enhances bone formation of UMR-106 cells and PKA, BMP and Hedgehog signalling pathways became activated. We also found that PACAP 6-38 did not act as an antagonist of PACAP signalling in UMR-106 cells.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Megjelenés:Journal of Molecular Neuroscience. - 54 : 3 (2014), p. 555-573. -
További szerzők:Matta Csaba (1980-) (molekuláris biológus, genetikus, angol szakfordító) Katona Éva (1986-) (molekuláris biológus) Somogyi Csilla (1983-) (biológus, angol-magyar szakfordító) Takács Roland Ádám (1985-) (molekuláris biológus, biokémikus) Hajdú Tibor (1988-) (általános orvos) Helgadottir, Solveig Lind Fodor János (1973-) (élettanász, biotechnológus) Csernoch László (1961-) (élettanász) Tóth Gábor Bakó Éva (1958-) (biokémikus) Reglődi Dóra (Idegtudományok) Tamás Andrea (Idegtudomány) (Pécs) Zákány Róza (1963-) (anatómus-, kötőszövetbiológus)
Pályázati támogatás:K 104984
OTKA
CNK80709
OTKA
TÁMOP-4.2.1.B-10/2KONV-2010-002
TÁMOP
TÁMOP-4.2.2.A-11/1/KONV-2012-0053
TÁMOP
TÁMOP-4.2.2.A-11/1/KONV-2012-0024
TÁMOP
TÁMOP-4.2.4.A/2-11-1-2012-0001
TÁMOP
Mecenatura Mec-9/2011
Egyéb
Internet cím:Szerző által megadott URL
DOI
Intézményi repozitóriumban (DEA) tárolt változat
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2.

001-es BibID:BIBFORM049124
035-os BibID:PMID:24333667
Első szerző:Juhász Tamás (biológus, orvosbiológus)
Cím:Mechanical loading stimulates chondrogenesis via the PKA/CREB-Sox9 and PP2A pathways in chicken micromass cultures / Tamás Juhász, Csaba Matta, Csilla Somogyi, Éva Katona, Roland Takács, Rudolf Ferenc Soha, István A. Szabó, Csaba Cserháti, Róbert Sződy, Zoltán Karácsonyi, Éva Bakó, Pál Gergely, Róza Zákány
Dátum:2014
ISSN:0898-6568
Megjegyzések:Biomechanical stimuli play important roles in the formation of articular cartilage during early foetal life, and optimal mechanical load is a crucial regulatory factor of adult chondrocyte metabolism and function. In this study, we undertook to analyse mechanotransduction pathways during in vitro chondrogenesis. Chondroprogenitor cells isolated from limb buds of 4-day-old chicken embryos were cultivated as high density cell cultures for 6days. Mechanical stimulation was carried out by a self-designed bioreactor that exerted uniaxial intermittent cyclic load transmitted by the culture medium as hydrostatic pressure and fluid shear to differentiating cells. The loading scheme (0.05Hz, 600Pa; for 30min) was applied on culturing days 2 and 3, when final commitment and differentiation of chondroprogenitor cells occurred in this model. The applied mechanical load significantly augmented cartilage matrix production and elevated mRNA expression of several cartilage matrix constituents, including collagen type II and aggrecan core protein, as well as matrix-producing hyaluronan synthases through enhanced expression, phosphorylation and nuclear signals of the main chondrogenic transcription factor Sox9. Along with increased cAMP levels, a significantly enhanced protein kinase A (PKA) activity was also detected and CREB, the archetypal downstream transcription factor of PKA signalling, exhibited elevated phosphorylation levels and stronger nuclear signals in response to mechanical stimuli. All the above effects were diminished by the PKA-inhibitor H89. Inhibition of the PKA-independent cAMP-mediators Epac1 and Epac2 with HJC0197 resulted in enhanced cartilage formation, which was additive to that of the mechanical stimulation, implying that the chondrogenesis-promoting effect of mechanical load was independent of Epac. At the same time, PP2A activity was reduced following mechanical load and treatments with the PP2A-inhibitor okadaic acid were able to mimic the effects of the intervention. Our results indicate that proper mechanical stimuli augment in vitro cartilage formation via promoting both differentiation and matrix production of chondrogenic cells, and the opposing regulation of the PKA/CREB-Sox9 and the PP2A signalling pathways is crucial in this phenomenon.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Doktori iskola
Mechanotransduction
Chondrocyte differentiation
Extracellular matrix
H89
Okadaic acid
Epac
Megjelenés:Cellular Signalling. - 26 : 3 (2014), p. 468-482. -
További szerzők:Matta Csaba (1980-) (molekuláris biológus, genetikus, angol szakfordító) Somogyi Csilla (1983-) (biológus, angol-magyar szakfordító) Katona Éva (1986-) (molekuláris biológus) Takács Roland Ádám (1985-) (molekuláris biológus, biokémikus) Soha Rudolf Ferenc (1986-) (fizikus) Szabó István András (1956-) (fizikus) Cserháti Csaba (1963-) (fizikus) Sződy Róbert Karácsonyi Zoltán (1975-) (ortopéd és baleseti sebész) Bakó Éva (1958-) (biokémikus) Gergely Pál (1947-) (biokémikus) Zákány Róza (1963-) (anatómus-, kötőszövetbiológus)
Pályázati támogatás:TÁMOP-4.2.2.A-11/1/KONV-2012-0025
TÁMOP
TÁMOP-4.2.2/B-10/1-2010-0024
TÁMOP
Molekuláris Orvostudomány Doktori Iskola
TÁMOP-4.2.2.A-11/1/KONV-2012-0036
TÁMOP
TÁMOP-4.2.4.A/2-11-1-2012-0001
TÁMOP
CNK80709
OTKA
Internet cím:Szerző által megadott URL
DOI
Intézményi repozitóriumban (DEA) tárolt változat
Borító:

3.

001-es BibID:BIBFORM013301
Első szerző:Juhász Tamás (biológus, orvosbiológus)
Cím:Optimalized transient transfection of chondrogenic primary cell cultures / Juhász Tamás, Matta Csaba, Mészár Zoltán, Nagy Georgina, Szíjgyártó Zsolt, Molnár Zsanett, Kolozsvári Bernadett, Bakó Éva, Zákány Róza
Dátum:2010
ISSN:1895-104X
Megjegyzések:We aimed to find a transfection method which provides high efficiency with minimal cytotoxic and/or apoptotic effects for gene transfer into multilayer primary chondrogenic cell cultures. The pEGFP-C1 plasmid was introduced into the cell culture and the efficiency of transformation quantified by GFP fluorescence; the resulting nucleofection was effective but resulted in severe apoptosis. Two liposomal reagents designed to allow transfection into adherent cells did not deliver the plasmids sufficiently and cartilage formation did not occur.In addition, a third liposomal compound, recommended for transfection into either adherent or suspension cell cultures, lead to acceptable transfection efficiency but no cartilage formation. When an amphiphilic reagent was used however, there was acceptable transfection efficiency as well as cartilage formation. The viability of the cells which were transfected using the amphiphilic reagent remained unaffected but proliferation was severely diminished, particularly in the presence of GFP. In addition, the amount of cartilage decreased when GFP was expressed, despite unchanged levels of mRNAs of sox9 and aggrecan core protein, factors reflecting on the efficiency of chondrogenesis. Overexpression of both the constitutively active delta and gamma isoforms of catalytic subunit of calcineurin, a protein phosphatase described as a positive regulator of chondrogenesis, decreased protein level of Sox9 and subsequent cartilage formation. In conclusion, we found that amphiphilic reagent applied prior to the adhesion of cells provides a useful means to transfer plasmids to primary differentiating chondrogenic cells.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Cartilage differentiation
Mesenchymal cell culture
Chondrifying primary cells
Transfection efficiency
Calcineurin overexpression
Megjelenés:Central European Journal of Biology. - 5 : 5 (2010), p. 572-584. -
További szerzők:Matta Csaba (1980-) (molekuláris biológus, genetikus, angol szakfordító) Mészár Zoltán Mihály (1977-) (agrármérnök) Nagy Georgina (1980-) (orvosbiológus) Szíjgyártó Zsolt (1978-) (vegyész) Molnár Zsanett (1982-) (analitikus) Kolozsvári Bernadett (1982-) (molekuláris biológus, genetikus) Bakó Éva (1958-) (biokémikus) Zákány Róza (1963-) (anatómus-, kötőszövetbiológus)
Internet cím:Szerző által megadott URL
DOI
Intézményi repozitóriumban (DEA) tárolt változat
Borító:
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