Muscarinic acetylcholine receptors in the brain of the zebrafish (Danio rerio) measured by radioligand binding techniques
Muscarinic acetylcholine receptors in the brain of the zebrafish (Danio rerio) measured by radioligand binding techniques
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DOI：
10.1016/j.cca.2004.03.002
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发表时间：
2004-04-01
影响因子：
3.9
通讯作者：
Messer, WS
中科院分区：
文献类型：
--
作者：
Williams, FE;Messer, WS
毒蕈碱乙酰胆碱受体 (mAChR) 在脊椎动物的学习、记忆和行为中发挥作用。我们通过放射性配体结合技术测量了斑马鱼（Danio rerio）大脑提取物中的毒蕈碱胆碱能受体水平。使用放射性配体[H-3]-苯苯甲酸奎宁环酯（QNB）进行饱和结合实验来确定受体数量以及几种激动剂和拮抗剂的相对亲和力。斑马鱼大脑受体的亲和力相对较高，K-d 为 40 +/- 5 pM。以 B-max 表示的受体数量为 63 +/- 16 fmol/mg 蛋白质。氧化震颤素和卡巴胆碱是毒蕈碱乙酰胆碱受体的激动剂，结合位移曲线表明有多个结合位点。此外，氧化震颤素的结合亲和力高于卡巴胆碱。斑马鱼脑内受体的拮抗剂效力分布被确定为阿托品远大于哌仑西平>对氟六氢西拉非尼多远大于奥替帕德。用斑马鱼大脑获得的结果优于在昆虫、鱼类和哺乳动物物种中发现的结果。综上所述，结合结果以及与哺乳动物系统的有利比较表明，斑马鱼可能为评估胆碱能系统在学习、记忆和行为中的作用提供有用的模型生物。 (C) 2004 Elsevier Inc. 保留所有权利。
Muscarinic acetylcholine receptors (mAChRs) play a role in learning, memory and behavior in vertebrate animals. We measured the muscarinic cholinergic receptor levels in extracts from zebrafish (Danio rerio) brain by radioligand binding techniques. Saturation binding experiments with the radioligand [H-3]-quinuclidinyl benzilate (QNB) were used to determine receptor number and relative affinity for several agonists and antagonists. Affinity at zebrafish brain receptors was relatively high with a K-d of 40 +/- 5 pM. The number of receptors, represented by B-max, was 63 +/- 16 fmol/mg protein. Oxotremorine and carbachol, agonists at muscarinic acetylcholine receptors, bound with displacement curves indicating multiple binding sites. In addition, oxotremorine bound with a higher affinity than did carbachol. The antagonist potency profile at zebrafish receptors in brain was determined to be atropine much greater than pirenzipine>p-fluoro-hexahydro-siladifenidol much greater than otenzepad. The results obtained with zebrafish brain compare favorably to those found in insect, fish and mammalian species. Taken together, the binding results and favorable comparisons to mammalian systems indicate that zebrafish may provide a useful model organism for evaluating the role of cholinergic systems in learning, memory and behavior. (C) 2004 Elsevier Inc. All rights reserved.