Fatima-Shad K, Barry P H
School of Physiology and Pharmacology, University of New South Wales, Sydney, Australia.
Proc Biol Sci. 1992 Nov 23;250(1328):99-105. doi: 10.1098/rspb.1992.0136.
GABA- and strychnine-sensitive glycine-activated currents in post-natal tissue-cultured hippocampal neurons were studied by using patch-clamp techniques. Current-voltage relations for both agonists in symmetrical Cl- solutions showed outward rectification. Strychnine-sensitive glycine-activated currents have not been studied in detail before in post-natal tissue-cultured hippocampal neurons. Partial desensitization of whole-cell currents was observed in symmetrical Cl- solutions during bath application of GABA- and glycine. In F-/Cl- solutions, both agonists gave a PF/Pcl value of about 0.06. The reversal potentials in mixtures of Cl- and SO4(2-) solutions were close to the equilibrium potentials of Cl- in the presence of both GABA and glycine. Single channels in inside-out excised patches with 2 mumol l-1 GABA and 5 mumol l-1 glycine in the pipette showed outward current rectification. The dose-response curves for GABA and glycine gave Kd values of 52 and 61 mumol l-1, respectively, and Hill coefficients close to 2. GABA and glycine binding were competitively blocked by their respective antagonists (bicuculline and strychnine). The similarities between GABA- and glycine-activated currents and the response in a combination of saturating concentrations of both GABA and glycine implied that the two agonists activated comparable numbers of anatomically distinct channels with very similar permeation properties.
运用膜片钳技术研究了出生后组织培养海马神经元中对γ-氨基丁酸(GABA)和士的宁敏感的甘氨酸激活电流。在对称氯离子溶液中,两种激动剂的电流-电压关系均表现出外向整流。此前尚未对出生后组织培养海马神经元中士的宁敏感的甘氨酸激活电流进行详细研究。在浴加GABA和甘氨酸期间,在对称氯离子溶液中观察到全细胞电流的部分脱敏现象。在氟离子/氯离子溶液中,两种激动剂的PF/Pcl值均约为0.06。在氯离子和硫酸根离子混合溶液中的反转电位接近在同时存在GABA和甘氨酸时氯离子的平衡电位。在移液管中含有2μmol/L GABA和5μmol/L甘氨酸的内面向外切除膜片中的单通道表现出外向电流整流。GABA和甘氨酸的剂量-反应曲线的Kd值分别为52和61μmol/L,希尔系数接近2。GABA和甘氨酸结合分别被其各自的拮抗剂(荷包牡丹碱和士的宁)竞争性阻断。GABA和甘氨酸激活电流之间的相似性以及在GABA和甘氨酸饱和浓度组合下的反应表明,这两种激动剂激活了数量相当的、具有非常相似通透特性的解剖学上不同的通道。