Sapp D W, Yeh H H
Department of Pharmacology, University of Connecticut Health Center, Farmington 06030, USA.
J Neurosci Res. 2000 May 15;60(4):504-10. doi: 10.1002/(SICI)1097-4547(20000515)60:4<504::AID-JNR9>3.0.CO;2-Y.
The gamma-aminobutyric acid (GABA) response profiles of IMR-32 human neuroblastoma cells were examined using whole-cell patch clamp and RT-PCR techniques. GABA activated a concentration-dependent and bicuculline-sensitive current, and RT-PCR revealed the expression of multiple GABA(A) receptor subunit mRNAs (alpha(1), alpha(3), alpha(4), beta(1), beta(3), gamma(2), and delta). A pharmacological profile of the GABA-induced current was derived using several subunit-selective agents. Diazepam, which requires the presence of a gamma subunit in order to modulate GABA(A) receptor-mediated responses, potentiated GABA-induced currents in a subset of IMR-32 cells. Two populations of GABA-activated currents were also evident based on sensitivity to modulation by zinc. Comparison of zinc- and diazepam-induced modulation of GABA-induced current responses in the same cells revealed an inverse correlation between these two modulators. No differences, however, were observed with the GABA(A) receptor modulators loreclezole, allopregnanolone, and pentobarbital. Thus, IMR-32 cells maintained in culture are heterogeneous in terms of expression of GABA(A) receptor isoforms.
采用全细胞膜片钳和逆转录聚合酶链反应(RT-PCR)技术检测了IMR-32人神经母细胞瘤细胞的γ-氨基丁酸(GABA)反应谱。GABA激活了浓度依赖性且对荷包牡丹碱敏感的电流,RT-PCR显示多种GABA(A)受体亚基mRNA(α(1)、α(3)、α(4)、β(1)、β(3)、γ(2)和δ)的表达。使用几种亚基选择性试剂得出了GABA诱导电流的药理学特征。地西泮需要γ亚基的存在才能调节GABA(A)受体介导的反应,它在一部分IMR-32细胞中增强了GABA诱导的电流。基于对锌调节的敏感性,还可明显看出两种GABA激活电流。在同一细胞中比较锌和地西泮对GABA诱导电流反应的调节作用,发现这两种调节剂之间呈负相关。然而,在GABA(A)受体调节剂氯雷唑、别孕烯醇酮和戊巴比妥方面未观察到差异。因此,培养的IMR-32细胞在GABA(A)受体亚型的表达方面是异质性的。