Bureau M, Laschet J, Bureau-Heeren M, Hennuy B, Minet A, Wins P, Grisar T
Laboratory of Neurochemistry, University of Liège, Belgium.
J Neurochem. 1995 Nov;65(5):2006-15. doi: 10.1046/j.1471-4159.1995.65052006.x.
GABAA receptors were characterized in cellular fractions isolated from adult bovine brain. The fraction enriched in cortical astrocytes is very rich in high-affinity binding sites for [3H]flunitrazepam and other "central-type" benzodiazepine ligands. The amount of specific [3H]flunitrazepam binding was more than five times higher in the glial fraction than in synaptosomal and perikaryal fractions. [3H]Flunitrazepam was displaced by low concentrations of clonazepam and other specific ligands for central GABAA receptors. Specific binding sites for GABA, flunitrazepam, barbiturates, and picrotoxin-like convulsants were characterized. Allosteric interactions between the different sites were typical of central-type GABAA receptors. The presence of alpha-subunit(s), as revealed by [3H]flunitrazepam photoaffinity labeling, was demonstrated in all brain fractions at molecular mas 51-53 kDa. Photoaffinity labeling was highest in the glial fraction. However, in primary cultured astrocytes from neonate rat cortex, no photoaffinity labeling was detected. Information obtained from astrocytes in culture should thus be taken with caution when extrapolated to differentiated astroglial cells. Our results actually show that, in mature brain, most of the fully pharmacologically active GABAA receptors are extrasynaptic and expressed in astroglia.
在从成年牛脑中分离出的细胞组分中对GABAA受体进行了表征。富含皮质星形胶质细胞的组分中富含[3H]氟硝西泮和其他“中枢型”苯二氮䓬配体的高亲和力结合位点。胶质细胞组分中特异性[3H]氟硝西泮结合量比突触体和核周组分中的高出五倍多。低浓度的氯硝西泮和其他中枢GABAA受体特异性配体可取代[3H]氟硝西泮。对GABA、氟硝西泮、巴比妥类药物和类似印防己毒素的惊厥剂的特异性结合位点进行了表征。不同位点之间的变构相互作用是中枢型GABAA受体的典型特征。通过[3H]氟硝西泮光亲和标记揭示的α亚基的存在,在所有脑组分中均在51-53 kDa的分子质量处得到证实。光亲和标记在胶质细胞组分中最高。然而,在新生大鼠皮质的原代培养星形胶质细胞中,未检测到光亲和标记。因此,当将从培养的星形胶质细胞获得的信息外推到分化的星形胶质细胞时应谨慎。我们的结果实际上表明,在成熟脑中,大多数具有完全药理活性的GABAA受体位于突触外并在星形胶质细胞中表达。