Huba R, Hofmann H D
Max-Planck-Institut für Hirnforschung, Frankfurt, Germany.
Vis Neurosci. 1991 Apr;6(4):303-14. doi: 10.1017/s0952523800006556.
A subpopulation of cells developing in dissociated neuronal cultures prepared from 8-day-old embryonic chick retinae can be identified as putative in vitro counterparts of GABAergic amacrine cells by immunocytochemical and autoradiographic markers and by their electrophysiological responses to transmitter agonists. In the present study, transmitter-gated conductances expressed by these neurons were examined using the whole-cell patch-clamp technique. At negative holding potentials, the excitatory amino acid agonists N-methyl-D-aspartate (NMDA), kainate quisqualate, and glutamate induced inward currents with reversal potentials close to 0 mV in most of the cells selected for recording. NMDA-evoked responses were selectively blocked by the noncompetitive inhibitor MK 801 and by Mg2+ (in a voltage-dependent manner) and were potentiated in the presence of submicromolar concentrations of glycine. Glutamate apparently interacted with both NMDA and non-NMDA type receptors. All cells tested responded to the inhibitory transmitters GABA and glycine. Both inhibitory agonists could be shown to activate chloride conductances. Responses to GABA and glycine were specifically inhibited in the presence of bicuculline and strychnine, respectively. Thus, GABAergic neurons in retinal cultures express at least two different excitatory amino acid receptors--NMDA and non-NMDA--and two different inhibitory amino acid receptors--the GABAA and the glycine receptor. The results demonstrate the ability of the cultured neurons to develop an apparently mature phenotype and contribute to the understanding of the functional properties of GABAergic amacrine cells in the vertebrate retina.
从8日龄鸡胚视网膜制备的解离神经元培养物中发育的一群细胞,可通过免疫细胞化学和放射自显影标记以及它们对递质激动剂的电生理反应,被鉴定为GABA能无长突细胞在体外的对应物。在本研究中,使用全细胞膜片钳技术检测了这些神经元表达的递质门控电导。在负的钳制电位下,兴奋性氨基酸激动剂N-甲基-D-天冬氨酸(NMDA)、海人藻酸、quisqualate和谷氨酸在大多数被选用于记录的细胞中诱导出内向电流,其反转电位接近0 mV。NMDA诱发的反应被非竞争性抑制剂MK 801和Mg2+(以电压依赖性方式)选择性阻断,并在亚微摩尔浓度的甘氨酸存在下增强。谷氨酸显然与NMDA型和非NMDA型受体都相互作用。所有测试的细胞都对抑制性递质GABA和甘氨酸有反应。两种抑制性激动剂都能激活氯离子电导。对GABA和甘氨酸的反应分别在荷包牡丹碱和士的宁存在时被特异性抑制。因此,视网膜培养物中的GABA能神经元表达至少两种不同的兴奋性氨基酸受体——NMDA和非NMDA——以及两种不同的抑制性氨基酸受体——GABAA和甘氨酸受体。这些结果证明了培养神经元形成明显成熟表型的能力,并有助于理解脊椎动物视网膜中GABA能无长突细胞的功能特性。