Autoradiography of receptor-activated G-proteins in post mortem human brain

被引:35
|
作者
Rodríguez-Puertas, R [1 ]
González-Maeso, J
Meana, JJ
Pazos, A
机构
[1] Univ Basque Country, Dept Pharmacol, Leioa, Bizkaia, Spain
[2] Univ Cantabria, Fac Med, Pharmacol Unit, Dept Physiol & Pharmacol, Santander 39011, Spain
关键词
S-35]GTP gamma S binding; alpha(2)-adrenoceptors; 5-HT1; receptors; mu-opioid receptors; cholinergic muscarinic receptors; cannabinoid receptors;
D O I
10.1016/S0306-4522(99)00527-8
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
The agonist-stimulated guanosine 5'-(gamma-[S-35]thio)triphosphate binding assay was used to anatomically localize receptor-activated G-proteins by autoradiography in post mortem human brain. The optimal conditions for guanosine 5'-(gamma-S-35]thio)triphosphate binding to human brain sections were established in post mortem samples of the prefrontal cortex, hippocampus, basal ganglia, brainstem and cerebellar cortex. An excess of GDP (2 mM) was required to decrease basal activity and obtain effective stimulation by specific agonists. guanosine 5'-(gamma-[S-35]Thio)triphosphate binding was increased after stimulation with specific agonists of different G-protein-coupled receptors. They include cannabinoid (WIN55212-2), mu-opioid ([D-Ala(2)N-MePh4,Gly(5)-ol]enkephalin), serotonin-1A [(+/-)-8-hydroxy-2-(di-n-propylamino)tetralin and serotonin-1B/1D (sumatriptan), cholinergic muscarinic receptors (carbachol) and alpha(2)-adrenoceptors (UK14304). Such stimulation reached 1458%, 440%, 188%, 219%, 61% and 339%, respectively, over the basal levels. In tissue sections, the use of the above-mentioned agonists (10(-4) M) showed patterns of anatomical distribution similar to those already described by receptor autoradiography, with high densities over the hippocampus (serotonin-1A receptors), cortex (alpha(2)-adrenoceptors) and striatum (mu-opioid receptors). The highest binding levels were reached with the cannabinoid receptor agonist in most of the analysed brain regions. Carbachol produced only moderate stimulation of those same regions. The blockage of agonist-stimulated guanosine 5'-(mu-[S-35]thio)triphosphate binding by selective antagonists verified that the effect was receptor mediated. This technique provides a method to identify modifications of the receptor-mediated activation of G-proteins in post mortem human brain with anatomical resolution. It also provides valuable information on the level of drug efficacy in the human species. (C) 2000 IBRO. Published by Elsevier Science Ltd.
引用
收藏
页码:169 / 180
页数:12
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