Intracellular application of TNF-alpha impairs cell to cell communication via gap junctions in glioma cells.
Human gliomas are the most common class of brain neoplasm. In order to better characterize their response to inflammation, we evaluated the influence of tumor necrosis factor alpha (TNF-alpha) on the coupling behaviour and the membrane resting potential (MRP) of glioma cells (F98 glioma cell line) compared to primary astrocytes. In contrast to cultured primary astrocytes which exhibited a profound inhibition of gap junction mediated intercellular communication (GJIC), extracellular exposure of TNF-alpha to F98 glioma cells gained no effect on the functional coupling. Whereas, intracellular application of TNF-alpha into the glioma cells elicited similar effects as those found in primary astrocytes indicating a compromised accessibility of the TNF-alpha receptor in F98 cells. Western blotting, immunocytochemical staining and real time RT PCR analysis revealed a differential expression and distribution of TNF-alpha receptor 1 (TNFR1) in the glioma cells. Connexin 43 (Cx43) is the major astrocytic gap junction protein which when phosphorylated has been shown to reveal altered gating properties. Here we show that TNF-alpha increases the level of phosphorylated Cx43 in primary astrocytes but not in the F98 glioma cells. Our observations could account for the decreased regulatory effects of TNF-alpha on GJIC of F98 glioma cells.
Haghikia
Aiden
A
Department of Neurology, St. Josef-Hospital Bochum, Ruhr-University Bochum, Gudrunstrasse 56, Bochum, Germany. Aiden.Haghikia@rub.de
Ladage
Kerstin
K
Lafênetre
Pauline
P
Haghikia
Arash
A
Hinkerohe
Daniel
D
Smikalla
Dirk
D
Haase
Claus G
CG
Dermietzel
Rolf
R
Faustmann
Pedro M
PM
D000818:Animals; D001253:Astrocytes / Q000378:metabolism* / Q000473:pathology; D001932:Brain Neoplasms / Q000378:metabolism* / Q000473:pathology; D002450:Cell Communication / Q000502:physiology; D018031:Connexin 43 / Q000378:metabolism; D017629:Gap Junctions / Q000378:metabolism*; D005910:Glioma / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007424:Intracellular Fluid / Q000378:metabolism; D051381:Rats; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
D018031:Connexin 43; D047888:Receptors, Tumor Necrosis Factor, Type I; D014409:Tumor Necrosis Factor-alpha
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