Jones S B, King L B
National Biological Service, Midwest Science Center, Columbia, MO 65201, USA.
Comp Biochem Physiol C Pharmacol Toxicol Endocrinol. 1995 Sep;112(1):43-50. doi: 10.1016/0742-8413(95)00070-4.
Muscarinic cholinergic receptors were measured by radioligand binding techniques in crude membrane particulate preparations of brain and atrial tissues from laboratory reared brook trout (Salvelinus fontinalis). The radioligand [3H]N-methyl scopolamine was used to determine number and affinity of receptors in saturation experiments. The affinity of the radioligand did not differ in brain and atrial preparations (96 +/- 8 and 60 +/- 4 pM, respectively). However, the number of binding sites was greater in atrium compared with brain (269 +/- 19 and 166 +/- 7 fmol/mg protein, respectively). The rank order of potency of competing drugs in inhibition experiments was similar for antagonists with atropine > or = scopolamine > pirenzepine. Pirenzepine, an M1-selective drug had a 3-fold higher affinity in brain than atrium. The agonists oxotremorine and carbachol each bound to two sites in both tissues. In contrast, pilocarpine bound to only one site in brain and two in atrial tissue. These results are compared with those observed in other nonmammalian species and discussed with reference to conservation of proteins that serve important cellular roles.
采用放射性配体结合技术,对实验室饲养的溪红点鲑(Salvelinus fontinalis)的脑和心房组织粗制膜颗粒制剂中的毒蕈碱胆碱能受体进行了测定。在饱和实验中,使用放射性配体[3H]N-甲基东莨菪碱来确定受体的数量和亲和力。放射性配体在脑制剂和心房制剂中的亲和力无差异(分别为96±8和60±4 pM)。然而,与脑相比,心房中的结合位点数更多(分别为269±19和166±7 fmol/mg蛋白质)。在抑制实验中,竞争性药物的效价排序与拮抗剂相似,阿托品≥东莨菪碱>哌仑西平。哌仑西平是一种M1选择性药物,在脑中的亲和力比在心房中高3倍。激动剂氧化震颤素和卡巴胆碱在两种组织中均与两个位点结合。相比之下,毛果芸香碱仅与脑中的一个位点结合,而与心房组织中的两个位点结合。将这些结果与在其他非哺乳动物物种中观察到的结果进行了比较,并参考了具有重要细胞功能的蛋白质的保守性进行了讨论。