Lambert D G, Ghataorre A S, Nahorski S R
Department of Pharmacology and Therapeutics, Leicester, U.K.
Eur J Pharmacol. 1989 Jun 8;165(1):71-7. doi: 10.1016/0014-2999(89)90771-1.
The present study examines the muscarinic receptor binding characteristics of parent human neuroblastoma (SK-N-SH) and its neuroblast (SH-SY5Y) and epithelial-like (SH-EP1) clones using [3H]methylscopolamine [( 3H]NMS). Specific [3H]NMS binding to intact SK-N-SH and SH-SY5Y cells was saturable with a Kd of 0.2 nM and Bmax of 100-150 fmol/mg protein. Specific [3H]NMS binding to whole cell preparations of SH-EP 1 could not be detected. Pharmacological analysis of the binding site both in whole cells and membranes of SK-N-SH are indicative of an homogeneous receptor population possessing low affinity for the M1-selective antagonist pirenzepine. The muscarinic receptors expressed by the neuroblast clone, SH-SY5Y were further characterized and shown to have the properties of an homogeneous M3 subtype with low affinity for the M1-selective antagonist pirenzepine and the M2-cardioselective AFDX-116 but high affinity for 4-diphenylacetoxy-N-methyl piperidine methiodide (4-DAMP). In conclusion the SH-SY5Y neuroblastoma should provide an important human neuronal cell model with which to define the regulation of post-receptor events driven by a single receptor population.
本研究使用[3H]甲基东莨菪碱（[3H]NMS）检测了人神经母细胞瘤亲本细胞（SK-N-SH）及其神经母细胞（SH-SY5Y）和上皮样细胞（SH-EP1）克隆的毒蕈碱受体结合特性。[3H]NMS与完整的SK-N-SH和SH-SY5Y细胞的特异性结合具有饱和性，解离常数（Kd）为0.2 nM，最大结合量（Bmax）为100 - 150 fmol/mg蛋白质。未检测到[3H]NMS与SH-EP1全细胞制剂的特异性结合。对SK-N-SH全细胞和细胞膜上结合位点的药理学分析表明，存在一群对M1选择性拮抗剂哌仑西平亲和力较低的同质受体。对神经母细胞克隆SH-SY5Y表达的毒蕈碱受体进行了进一步表征，结果显示其具有同质M3亚型的特性，对M1选择性拮抗剂哌仑西平和M2心脏选择性AFDX-116亲和力较低，但对4-二苯基乙酰氧基-N-甲基哌啶甲碘化物（4-DAMP）亲和力较高。总之，SH-SY5Y神经母细胞瘤应能提供一个重要的人类神经元细胞模型，用以确定由单一受体群体驱动的受体后事件的调节机制。
