Benzodiazepines block alpha2-containing inhibitory glycine receptors in embryonic mouse hippocampal neurons.
Inhibitory glycine receptors (GlyRs) in the mammalian cortex probably contribute to brain development and to maintaining tonic inhibition. Given their presence throughout the cortex, their modulation likely has important physiological consequences. Although benzodiazepines potentiate gamma-aminobutyric acidA receptors (GABAARs), they may also modulate GlyRs because binding studies initially suggested that they act at GlyRs. Furthermore, their diminished ability to potentiate neonatal GABAARs suggests that they may exert their beneficial clinical effects at another site in the developing brain. Therefore we examined the effect of benzodiazepines on whole cell currents mediated by GlyRs in cultured embryonic mouse hippocampal neurons. First, we determined the GlyR subunit composition in this preparation. Glycine, beta-alanine, and taurine activate strychnine-sensitive chloride currents in a dose-dependent manner. Maximal concentrations of the three agonists produce equal, nonadditive responses as expected of full agonists. The pharmacological properties of the GlyR currents including their pattern of modulation by picrotoxinin, picrotin, and tropisetron indicate that GlyRs consist of alpha2beta heteromers and alpha2 homomers. Reverse transcriptase polymerase chain reaction (RTPCR) studies confirmed the presence of alpha2 and beta subunits. Second, we found that micromolar concentrations of some benzodiazepines, including chlordiazepoxide and nitrazepam, inhibit GlyR currents. Nitrazepam inhibition of GlyRs is noncompetitive, is not voltage dependent, and does not reflect enhanced desensitization. Thus benzodiazepines allosterically inhibit alpha2-containing GlyRs in embryonic mouse hippocampal neurons via a "low"-affinity site.
Thio
Liu Lin
LL
Department of Neurology, Washington University School of Medicine, St. Louis, Missouri, USA. thio@kids.wustl.edu
Shanmugam
Ananth
A
Isenberg
Keith
K
Yamada
Kelvin
K
D000818:Animals; D001569:Benzodiazepines / Q000494:pharmacology*; D018929:Cell Culture Techniques; D005998:Glycine / Q000494:pharmacology; D006624:Hippocampus / Q000737:chemistry / Q000187:drug effects* / Q000196:embryology / Q000502:physiology; D051379:Mice; D009474:Neurons / Q000737:chemistry / Q000187:drug effects* / Q000502:physiology; D018408:Patch-Clamp Techniques; D018009:Receptors, Glycine / Q000737:chemistry* / Q000187:drug effects*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013654:Taurine / Q000494:pharmacology; D015091:beta-Alanine / Q000494:pharmacology
D018009:Receptors, Glycine; D015091:beta-Alanine; D001569:Benzodiazepines; D013654:Taurine; D005998:Glycine