Distinct kinetic binding propeties of N‐[3H]‐methylscopolamine afford differential labeling and localization of M1, M2, and M3 muscarinic receptor subtypes in primate brain
Distinct kinetic binding propeties of N‐[3H]‐methylscopolamine afford differential labeling and localization of M1, M2, and M3 muscarinic receptor subtypes in primate brain
复制标题
N-[3H]-甲基东莨菪碱的独特动力学结合特性为灵长类动物脑中 M1、M2 和 M3 毒蕈碱受体亚型提供了差异标记和定位
作者：
D. Flynn;D. Mash
根据平衡结合和动力学结合数据，已假定哺乳动物脑中存在三类毒蕈碱受体。然而，单独的平衡结合试验并不能清楚地证明脑中假定的M1、M2和M3受体亚型的定位，因为几乎所有的毒蕈碱拮抗剂都具有亲和力。在本研究中，恒河猴脑中M1、M2和M3受体亚型选择性占据的条件是基于N-[3 H]-甲基东莨菪碱（NMS）与A9 L转染细胞中表达的克隆m1-m4毒蕈碱受体亚型的不同动力学和平衡结合特性。根据以下策略对灵长类动物脑中的M1、M2和M3毒蕈碱受体亚型进行定量放射自显影。用非饱和浓度的[3 H]-哌仑西平直接标记M1（m1）受体亚型。在与0.3 μM哌仑西平预孵育以封闭m1、m3和m4位点后，通过孵育（由短的2分钟[3 H]-NMS脉冲组成）标记M2（m2）亚型。通过与0.5 nM未标记NMS预孵育以部分封闭m1、m2和m4位点，与0.5 nM [3 H]-NMS平衡，然后在1 μM阿托品存在下进行60分钟示踪剂解离，实现M3（m3）受体（亚型）的选择性占据。
Three classes of muscarinic receptors in mammalian brain have been postulated on the basis of equilibrium and kinetic binding data. However, equilibrium binding assays alone have not permitted a clear demonstration of the localization of putative M1, M2, and M3 receptor subtypes in the brain because of the overlaping affinities of virtually all muscarinic antagonists. In the present study, the conditions for selective occupancy of the M1, M2, and M3 receptor subtypes in the brain of the rhesus monkey were based on the distinct kinetic and equilibrium binding properties of N‐[3H]‐methylscopolamine (NMS) at cloned m1–m4 muscarinic receptor subtypes expressed in A9L transfected cells. Quantitative autoradiography of the M1, M2, and M3 muscarinic receptor subtypes in the primate brain was performed according to the following strategy. The M1 (m1) receptor subtype was labeled directly with a non‐saturating concentration of [3H]‐pirenzepine. The M2 (m2) subtype was labeled by incubations consisting of short, two minute pulses of [3H]‐NMS after a preincubation with 0.3 μM pirenzepine to occlude m1, m3, and m4 sites. Selective occupancy of the M3 (m3) receptor (subtype) was achieved by pre‐incubation with 0.5 nM unlabeled NMS to partially occlude the m1, m2, and m4 sites, equilibrium with 0.5 nM [3H]‐NMS, followed by a 60 minute tracer dissociation in the presence of 1 μM atropine.
登录
查看更多内容
针对 m3 毒蕈碱受体的抗血清的开发：m3 受体在大鼠组织和克隆细胞系中的分布。
影响因子：
3.6
作者：
Wall,SJ;Yasuda,RP;Li,M;Wolfe,BB
通讯作者：
Wolfe,BB
使用 m2 受体选择性抗血清观察大鼠脑中 m2 毒蕈碱受体的分布。
影响因子：
3.6
作者：
Li,M;Yasuda,RP;Wall,SJ;Wellstein,A;Wolfe,BB
通讯作者：
Wolfe,BB
使用融合蛋白生产针对 m1 毒蕈碱受体的选择性抗血清：大鼠脑中 m1 受体的分布。
影响因子：
3.6
作者：
Wall,SJ;Yasuda,RP;Hory,F;Flagg,S;Martin,BM;Ginns,EI;Wolfe,BB
通讯作者：
Wolfe,BB
大鼠脑中的毒蕈碱 M2 受体用 [3H] AF-DX 384 标记。
DOI：
--
发表时间：
1991
期刊：
Research communications in chemical pathology and pharmacology
影响因子：
--
作者：
Castoldi,AF;Fitzgerald,B;Manzo,L;Tonini,M;Costa,LG
通讯作者：
Costa,LG
影响因子：
56.9
作者：
MASH, DC;FLYNN, DD;POTTER, LT
通讯作者：
POTTER, LT