Chieng B, Bekkers J M
Division of Neuroscience, John Curtin School of Medical Research, Australian National University, Canberra ACT 0200, Australia.
Br J Pharmacol. 2001 Jun;133(3):337-44. doi: 10.1038/sj.bjp.0704072.
The pharmacological effects of opioid- and adenosine-receptor agonists on neural signalling were investigated by measuring drug actions on barium current flowing through calcium channels in acutely-dissociated neurons of the rat nucleus accumbens (NAc). Under whole-cell voltage clamp, opioids acted via mu, but not delta or kappa, receptors to partially inhibit barium current. Mean inhibition was 35+/-2% (+/-s.e.mean, n = 33) for methionine-enkephalin and 37+/-1% (n = 65) for the selective mu receptor agonist DAMGO, both measured at saturating agonist concentrations in neurons with diameter > or = 20 microm. EC(50) for DAMGO was 100 nM. Perfusion of naloxone reversed the current inhibition by DAMGO. Adenosine also partially inhibited barium current in these neurons. Mean inhibition was 28+/-2% (n = 29) for adenosine and 33+/-3% (n = 27) for the selective A1 receptor agonist N(6)CPA, both at saturating concentrations in neurons with diameter > or = 20 microm. EC(50) for N(6)CPA was 34 nM. Adenosine inhibition was reversed by perfusion of an A1 receptor antagonist, 8-cyclopentyl-1,3-dipropylxanthine, while the selective A2A receptor agonist, CGS 21680, had no effect. Inhibition by opioids and adenosine was mutually occlusive, suggesting a converging pathway onto calcium channels. These actions involved a G-protein-coupled mechanism, as demonstrated by the partial relief of inhibition by strong depolarization and by the application of N-ethylmaleimide or GTP-gamma-S. Inhibition of barium current by opioids had their greatest effect in large neurons, that is, in presumed interneurons. In contrast, opioid inhibition in neurons with diameter < or = 15 microm was 11+/-2% (n = 26) for methionine-enkephalin and 11+/-4% (n = 17) for DAMGO, both measured at saturating agonist concentrations. Adenosine inhibition in neurons with diameter < or = 15 microm was 22+/-5% (n = 9). These results implicate the interneurons as a locus for the modulation of the excitability of projection neurons in the NAc during the processes of addiction and withdrawal.
通过测量阿片类和腺苷受体激动剂对大鼠伏隔核(NAc)急性解离神经元中流经钙通道的钡电流的作用,研究了它们对神经信号传导的药理作用。在全细胞电压钳制下,阿片类药物通过μ受体而非δ或κ受体发挥作用,部分抑制钡电流。在直径≥20微米的神经元中,以饱和激动剂浓度测量时,甲硫氨酸脑啡肽的平均抑制率为35±2%(±标准误均值,n = 33),选择性μ受体激动剂DAMGO的平均抑制率为37±1%(n = 65)。DAMGO的EC50为100 nM。纳洛酮灌注可逆转DAMGO对电流的抑制作用。腺苷也部分抑制这些神经元中的钡电流。在直径≥20微米的神经元中,以饱和浓度测量时,腺苷的平均抑制率为28±2%(n = 29),选择性A1受体激动剂N6CPA的平均抑制率为33±3%(n = 27)。N6CPA的EC50为34 nM。A1受体拮抗剂8-环戊基-1,3-二丙基黄嘌呤灌注可逆转腺苷的抑制作用,而选择性A2A受体激动剂CGS 21680则无作用。阿片类药物和腺苷的抑制作用相互重叠,表明存在一条汇聚到钙通道的途径。这些作用涉及G蛋白偶联机制,强去极化以及应用N-乙基马来酰亚胺或GTP-γ-S可部分缓解抑制作用,证明了这一点。阿片类药物对钡电流的抑制作用在大型神经元中效果最为显著,即推测为中间神经元。相比之下,在直径≤15微米的神经元中,以饱和激动剂浓度测量时,甲硫氨酸脑啡肽的抑制率为11±2%(n = 26),DAMGO的抑制率为11±4%(n = 17)。在直径≤15微米的神经元中,腺苷的抑制率为22±5%(n = 9)。这些结果表明,在成瘾和戒断过程中,中间神经元是调节NAc中投射神经元兴奋性的一个位点。