Functional GABA(A) receptors on human glioma cells.
Glioma cells in acute slices and in primary culture, and glioma-derived human cell lines were screened for the presence of functional GABA(A) receptors. Currents were measured in whole-cell voltage clamp in response to gamma-aminobutyric acid (GABA). While cells from the most malignant glioma, the glioblastoma multiforme, did not respond to GABA, an inward current (under our experimental conditions with high Cl- concentration in the pipette) was induced in gliomas of lower grades, namely in 71% of oligodendroglioma cells and in 62% of the astrocytoma cells. Glioma cell lines did not express functional GABA(A) receptors, irrespective of the malignancy of the tumour they originate from. The currents elicited by application of GABA were due to activation of GABA(A) receptors; the specific agonist muscimol mimicked the response, the antagonists bicuculline and picrotoxin blocked the GABA-activated current and the benzodiazepine receptor agonist flunitrazepam augmented the GABA-induced current and the benzodiazepine inverse agonist DMCM decreased the GABA current. Cells were heterogeneous with respect to the direction of the current flow as tested in gramicidin perforated patches: in some cells GABA hyperpolarized the membrane, while in the majority it triggered a depolarization. Moreover, GABA triggered an increase in [Ca2+]i in the majority of the tumour cells due to the activation of Ca2+ channels. Our results suggest a link between the expression of GABA receptors and the growth of glioma cells as the disappearance of functional GABA(A) receptors parallels unlimited growth typical for malignant tumours and immortal cell lines.
Labrakakis
C
C
Department of Cellular Neurosciences, Max Delbrück Center for Molecular Medicine (MDC), Berlin, Germany.
Patt
S
S
Hartmann
J
J
Kettenmann
H
H
D000900:Anti-Bacterial Agents; D002118:Calcium / Q000493:pharmacokinetics; D002121:Calcium Channel Blockers / Q000494:pharmacology; D005445:Flunitrazepam / Q000494:pharmacology; D018755:GABA Agonists / Q000494:pharmacology; D018757:GABA Modulators / Q000494:pharmacology; D005909:Glioblastoma / Q000737:chemistry*; D006096:Gramicidin; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D009118:Muscimol / Q000494:pharmacology; D009837:Oligodendroglioma / Q000737:chemistry*; D018408:Patch-Clamp Techniques; D010852:Picrotoxin / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology; D011963:Receptors, GABA-A / Q000032:analysis* / Q000502:physiology*; D014407:Tumor Cells, Cultured / Q000737:chemistry; D014700:Verapamil / Q000494:pharmacology; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D002118:Calcium; D002121:Calcium Channel Blockers; D005445:Flunitrazepam; D018755:GABA Agonists; D018757:GABA Modulators; D006096:Gramicidin; D009118:Muscimol; D010852:Picrotoxin; D011188:Potassium; D011963:Receptors, GABA-A; D014700:Verapamil; D005680:gamma-Aminobutyric Acid