European Journal of Pharmacology: Molecular Pharmacology
Endogenous [3H]flunitrazepam binding in human embryonic kidney cell line 293
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Cited by (38)
Novel positive allosteric modulators of GABAA receptors: Do subtle differences in activity at α1 plus α5 versus α2 plus α3 subunits account for dissimilarities in behavioral effects in rats?
2010, Progress in Neuro-Psychopharmacology and Biological PsychiatryDifferential effects of diazepam treatment and withdrawal on recombinant GABA<inf>A</inf> receptor expression and functional coupling
2008, Brain ResearchCitation Excerpt :In withdrawal studies, the medium containing diazepam was after 48 h replaced with drug-free medium for the next 24 h. Cell membranes were prepared mainly as described by Fuchs et al. (1995). The cells were washed, harvested by scraping into ice-cold phosphate-buffer saline (PBSA) and centrifuged at 12,000 ×g for 10 min, at 4 °C.
PWZ-029, a compound with moderate inverse agonist functional selectivity at GABA<inf>A</inf> receptors containing α5 subunits, improves passive, but not active, avoidance learning in rats
2008, Brain ResearchCitation Excerpt :The non-selective antagonist flumazenil was donated from F. Hoffman-La Roche (Basel, Switzerland) and the non-selective inverse agonist DMCM was purchased from Research Biochemicals Incorporated (Natick, MA, USA). Cloning of GABAA receptor subunits α1, β3 and γ2 into pCDM8 expression vectors (Invitrogen, CA) has been described elsewhere (Fuchs et al., 1995). cDNAs for subunits α2, α3 and α5 were gifts from P. Malherbe and were subcloned into pCI-vector.
Channel opening by anesthetics and GABA induces similar changes in the GABA<inf>A</inf> receptor M2 segment
2007, Biophysical JournalCitation Excerpt :Of note, Lynch and co-workers reported that in HEK293 cells they did not observe Cu:phen-induced increases in holding current (56). It is possible that the difference may arise because some HEK cells lines have been shown to express significant levels of endogenous wild-type β-subunit (57,58). Given the β-subunit dependence that we observed at the 6′ level, the coexpression of endogenous wild-type β-subunits would have a major impact on the results.
Chronic treatment with flumazenil enhances binding sites for convulsants at recombinant α<inf>1</inf>β<inf>2</inf>γ<inf>2S</inf> GABA <inf>A</inf> receptors
2005, Biomedicine and PharmacotherapyCitation Excerpt :In addition, the potency of drugs to enhance or inhibit [3H]flunitrazepam binding mainly corresponded to that observed for the modulation of the binding of [3H]flunitrazepam to the native type 1 benzodiazepine binding sites [30]. Fuchs et al. [8] identified the presence of low amounts of α1-, β3- and γ2-subunit mRNAs as well as endogenous [3H]flunitrazepam binding in untransfected HEK 293 cells. However, when the binding to the membranes of untransfected cells was performed under conditions used for transfected cells, no specific binding of either [3H]TBOB or [3H]flunitrazepam could be detected (our unpublished results), suggesting that endogenous subunits of GABAA receptor do not interfere with the binding of these labeled ligands to HEK 293 cells stably expressing recombinant α1β2γ2S GABAA receptor subunits.
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