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001-es BibID:BIBFORM015613
035-os BibID:11195086
Első szerző:Zákány Róza (anatómus-, kötőszövetbiológus)
Cím:Okadaic acid-induced inhibition of protein phosphatase 2A enhances chondrogenesisin chicken limb bud micromass cell cultures / Róza Zákány, Éva Bakó, Szabolcs Felszeghy, Krisztina Holló, Margit Balázs, Helga Bárdos, Pál Gergely, László Módis
Dátum:2001
ISSN:0340-2061 (Linking)
Megjegyzések:The role of major cellular serine/threonine-specific protein phosphatases,protein phosphatase 1 and 2A, was investigated during chicken cartilagedifferentiation under in vitro conditions. Activity of protein phosphatase 2Adecreased parallel to differentiation of chondrogenic cells, whereas activity ofprotein phosphatase 1 remained unchanged as assayed in the supernatants of thehomogenised chicken limb bud micromass cell cultures. When okadaic acid, a potentinhibitor of protein phosphatase 1 and 2A was applied in 20 nM concentration for4 h during the second and third culturing days, it significantly increased thesize of metachromatic cartilage areas measured in 6-day-old colonies. Followingokadaic acid treatments, a significant inhibition in the activity of proteinphosphatase 2A was found, while the activity of protein phosphatase 1 wasunaffected as measured an days 2 and 3. TRITC-phalloidin labelling demonstratedthat okadaic acid disorganised actin filaments and induced rounding ofchondrogenic cells. This deterioration of actin filaments was reversible.Electron microscopy and biochemical analysis of colonies revealed that theultrastructure and major components of cartilage matrix remained unchanged underthe effect of okadaic acid. Okadaic acid-treatment applied to cultures containingpredominantly differentiated chondrocytes (after day 4) did not influence thecartilage formation. 3H-thymidine and bromodeoxyuridine incorporation-assaysdemonstrated enhanced cell proliferation in the okadaic acid-treated coloniescompared to that of the untreated ones. Our results indicate, for the first time,that protein phosphatase 2A is involved in the regulation of chondrogenesis.Inhibition of protein phosphatase 2A with okadaic acid may result in increasedchondrogenesis via modulation of proliferation and cytoskeletal organisation, aswell as via alteration of protein kinase A-signaling pathway of the chondrogeniccells.
Tárgyszavak:Orvostudományok Klinikai orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
78111-17-8 (Okadaic Acid)
EC 3.1.3.16 (Phosphoprotein Phosphatases)
EC 3.1.3.16 (Protein Phosphatase 1)
EC 3.1.3.16 (Protein Phosphatase 2)
Animals
Cartilage/*embryology/metabolism/ultrastructure
Cell Differentiation/drug effects/physiology
Cell Division/drug effects/physiology
Cells, Cultured/drug effects/metabolism/ultrastructure
Chick Embryo
Chondrocytes/drug effects/*metabolism/ultrastructure
Chondrogenesis/drug effects/*physiology
Cytoskeleton/drug effects/metabolism/ultrastructure
Dose-Response Relationship, Drug
Limb Buds/*embryology/metabolism/ultrastructure
Microfilaments/drug effects/metabolism/ultrastructure
Okadaic Acid/*pharmacology
Phosphoprotein Phosphatases/drug effects/*metabolism
Phosphorylation/drug effects
Protein Phosphatase 1
Protein Phosphatase 2
Signal Transduction/drug effects/physiology
egyetemen (Magyarországon) készült közlemény
Megjelenés:Anatomy and Embryology. - 203 : 1 (2001), p. 23-34. -
További szerzők:Bakó Éva (1958-) (biokémikus) Felszeghy Szabolcs Béla (1972-) (fogorvos, anatómus, kötőszövetbiológus) Holló Krisztina (1967-) (vegyész) Balázs Margit (1952-) (sejtbiológus, molekuláris genetikus) Bárdos Helga (1969-) (megelőző orvostan és népegészségtan szakorvos) Gergely Pál (1947-) (biokémikus) Módis László (1939-) (anatómus, kötőszövetbiológus)
Internet cím:Intézményi repozitóriumban (DEA) tárolt változat
elektronikus változat
DOI
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2.

001-es BibID:BIBFORM014363
Első szerző:Zákány Róza (anatómus-, kötőszövetbiológus)
Cím:Protein Phosphatase 2A Is Involved in the Regulation of Protein Kinase A Signaling Pathway during in Vitro Chondrogenesis / Róza Zákány, Kornélia Szűcs, Éva Bakó, Szabolcs Felszeghy, Gabriella Czifra, Tamás Bíró, László Módis, Pál Gergely
Dátum:2002
ISSN:0014-4827
Megjegyzések:aWe have evaluated the importance of the Ser/Thr protein phosphorylation anddephosphorylation for chondrogenesis in high-density chicken limb bud mesenchymalcell cultures (HDCs) by using H89, a cell-permeable protein kinase inhibitor, andokadaic acid (OA), a phosphoprotein phosphatase (PP)-specific inhibitor molecule.When 20 nM OA was applied to the HDCs on Days 2 and 3 of culturing, itsignificantly inhibited protein phosphatase 2A (PP2A), enhanced cartilageformation, and elevated the activity of cAMP-dependent protein kinase (PKA).Application of 20 microM H89 significantly decreased the activity of PKA andblocked the chondrogenesis in HDCs. Furthermore, OA enhanced cartilage formationand elevated the suppressed activity of PKA even in the H89-pretreated HDCs.cGMP-dependent protein kinase was not detected in HDCs, while protein kinase Cmu(PKCmu), which is also inhibited by nanomolar concentrations of H89, was presentthroughout the culturing period. Neither OA nor H89 influenced the expression ofthe catalytic subunit of PKA or the cAMP response element binding protein, CREB.However, a significantly elevated amount of Ser-133-phosphorylated-CREB (P-CREB)was detected following addition of OA, while H89 treatment resulted in a decreaseof the amount of P-CREB. Our results demonstrate that PP2A plays a role in theregulation of the PKA signaling pathway and that the phosphorylation level ofCREB is influenced by the activity of both enzymes during in vitrochondrogenesis.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
a0 (Cyclic AMP Response Element-Binding Protein)
0 (Enzyme Inhibitors)
0 (Isoquinolines)
0 (Sulfonamides)
127243-85-0 (N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide)
56-45-1 (Serine)
72-19-5 (Threonine)
78111-17-8 (Okadaic Acid)
EC 2.7.11.11 (Cyclic AMP-Dependent Protein Kinases)
EC 3.1.3.16 (Phosphoprotein Phosphatases)
EC 3.1.3.16 (Protein Phosphatase 2)
Animals
Cartilage/drug effects/embryology/metabolism
Cell Differentiation/drug effects/physiology
Cell Division/drug effects/physiology
Cells, Cultured
Chick Embryo
Chondrocytes/drug effects/metabolism
Chondrogenesis/drug effects/*physiology
Cyclic AMP Response Element-Binding Protein/metabolism
Cyclic AMP-Dependent Protein Kinases/antagonists & inhibitors/*metabolism
Dose-Response Relationship, Drug
Enzyme Inhibitors/*pharmacology
Isoquinolines/pharmacology
Limb Buds/embryology/metabolism
Okadaic Acid/pharmacology
Phosphoprotein Phosphatases/drug effects/*metabolism
Phosphorylation
Protein Phosphatase
Serine/chemistrySignal Transduction
Sulfonamides
Threonine/chemistry
Time Factors
egyetemen (Magyarországon) készült közlemény
Megjelenés:Experimental Cell Research. - 275 : 1 (2002), p. 1-8. -
További szerzők:Szűcs Kornélia (1945-) (biokémikus) Bakó Éva (1958-) (biokémikus) Felszeghy Szabolcs Béla (1972-) (fogorvos, anatómus, kötőszövetbiológus) Czifra Gabriella (1975-) (élettanász) Bíró Tamás (1968-) (élettanász) Módis László (1939-) (anatómus, kötőszövetbiológus) Gergely Pál (1947-) (biokémikus)
Internet cím:Szerző által megadott URL
DOI
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