Effects of ethanol on recombinant glycine receptors expressed in mammalian cell lines.
We examined the effects of acute ethanol exposure on recombinant human glycine receptors transiently transfected into HEK 293 cells and stably transfected into Ltk- fibroblast-like cells. In our study of the effects of ethanol, we used the whole-cell patch-clamp configuration. Relatively low concentrations of ethanol (25 mM and 50 mM) did not affect glycine-gated currents in any of the cell lines studied. Higher concentrations of ethanol (100 mM and 200 mM) significantly potentiated glycine responses only in stably transfected Ltk- cells expressing alpha1 and alpha2 subunits and in HEK 293 cells transiently expressing alpha2 subunits. Cells stably expressing alpha1 versus alpha2 glycine receptors were modulated equally by ethanol. Both glycine alpha1 and glycine alpha1beta receptors transiently expressed in HEK 293 cells were insensitive to all concentrations of ethanol tested; however, there was a trend toward potentiation at 100 and 200 mM ethanol concentrations. A population of cells (41-87%) that was sensitive to the potentiating effects of 100 and 200 mM ethanol (defined as more than 10% potentiation) was identified in both cell lines tested. In these sensitive cells, ethanol (100 and 200 mM) produced significant potentiation, independent of the cell line and the glycine receptor subunit tested. Together with published results from studies with Xenopus oocytes, these data indicate that the sensitivity of recombinant glycine receptors to ethanol depends upon the expression system.
Valenzuela
C F
CF
Department of Pharmacology, University of Colorado Health Sciences Center, Denver 80262, USA.
Cardoso
R A
RA
Wick
M J
MJ
Weiner
J L
JL
Dunwiddie
T V
TV
Harris
R A
RA
D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D000431:Ethanol / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007473:Ion Channels / Q000187:drug effects; D018408:Patch-Clamp Techniques; D010766:Phosphorylation / Q000187:drug effects; D018009:Receptors, Glycine / Q000235:genetics*; D011994:Recombinant Proteins
D007473:Ion Channels; D018009:Receptors, Glycine; D011994:Recombinant Proteins; D000431:Ethanol