Laboratorio de Neurobiología Celular y Molecular, Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres" (INGEBI), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Vuelta de Obligado 2490, Ciudad Autónoma de Buenos Aires CP 1428, Argentina.
Laboratorio de Neurobiología Celular y Molecular, Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres" (INGEBI), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Vuelta de Obligado 2490, Ciudad Autónoma de Buenos Aires CP 1428, Argentina.
Eur J Pharmacol. 2014 Nov 15;743:24-30. doi: 10.1016/j.ejphar.2014.09.017. Epub 2014 Sep 22.
GABA(A) receptors (GABA(A)Rs) are ligand-gated ion channels that mediate inhibitory neurotransmission in the central nervous system (CNS). They are members of the Cys-loop receptor family and display marked structural and functional heterogeneity. Many GABA(A)Rs receptor subtypes are allosterically modulated by benzodiazepines (BDZs), which are drugs extensively used as anxiolytics, sedative-hypnotics and anticonvulsants. One high-affinity site and at least three additional low-affinity sites for BDZ recognition have been identified in several heteromeric and homomeric variants of the GABA(A)Rs (e.g.: α1β2γ2, α1β2/3, β3, etc.). However, the modulation of homomeric GABA(A)ρRs by BDZs was not previously revealed, and these receptors, for a long a time, were assumed to be fully insensitive to the actions of these drugs. In the present study, human homomeric GABA(A)ρ1 receptors were expressed in Xenopus oocytes and GABA-evoked responses electrophysiologically recorded in the presence or absence of BDZs. GABA(A)ρ1 receptor-mediated responses were modulated by diazepam and 4'-chlorodiazepam in the micromolar range, in a concentration-dependent, voltage-independent and reversible manner. Diazepam produced potentiating effects on GABA-evoked Cl(-) currents and 4'-Cl diazepam induced biphasic effects depending on the GABA concentration, whereas Ro15-4513 and alprazolam were negative modulators. BDZ actions were insensitive to flumazenil. Other BDZs showed negligible activity at equivalent experimental conditions. Our results suggest that GABA(A)ρ1 receptor function can be selectively and differentially modulated by BDZs.
GABA(A) 受体 (GABA(A)Rs) 是配体门控离子通道,在中枢神经系统 (CNS) 中介导抑制性神经传递。它们是 Cys 环受体家族的成员,表现出显著的结构和功能异质性。许多 GABA(A)Rs 受体亚型都被苯二氮䓬类药物 (BDZs) 变构调节,BDZs 是广泛用作抗焦虑药、镇静催眠药和抗惊厥药的药物。在几种 GABA(A)Rs 的异源和同源变体中(例如:α1β2γ2、α1β2/3、β3 等),已经鉴定出一个高亲和力位点和至少三个额外的低亲和力位点用于 BDZ 识别。然而,BDZs 对同源 GABA(A)ρRs 的调制以前没有被揭示,并且这些受体很长一段时间以来被认为对这些药物的作用完全不敏感。在本研究中,在非洲爪蟾卵母细胞中表达了人同源 GABA(A)ρ1 受体,并在存在或不存在 BDZs 的情况下用电生理学记录 GABA 诱发的反应。GABA(A)ρ1 受体介导的反应在微摩尔范围内被地西泮和 4'-氯地西泮以浓度依赖性、电压非依赖性和可逆转的方式调节。地西泮对 GABA 诱发的 Cl(-)电流产生增效作用,而 4'-Cl 地西泮则根据 GABA 浓度产生双相作用,而 Ro15-4513 和阿普唑仑则是负调节剂。BDZ 作用对地西泮不敏感。在等效实验条件下,其他 BDZs 表现出微不足道的活性。我们的结果表明,BDZs 可以选择性和差异化地调节 GABA(A)ρ1 受体功能。