Stereospecific interaction between narcotic analgesics and a synaptic plasm a membrane fraction of rat cerebral cortex.
Stereospecific interaction between narcotic analgesics and a synaptic plasm a membrane fraction of rat cerebral cortex.
复制标题
麻醉镇痛药与大鼠大脑皮层膜部分突触质之间的立体特异性相互作用。
DOI：
10.1111/j.1600-0773.1973.tb01477.x
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
L. Terenius
中科院分区：
文献类型：
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作者：
L. Terenius
了解麻醉镇痛药受体的亚细胞定位对于理解其作用机制非常重要。二氢吗啡-3H (DHM) 的特异性摄取已在豚鼠回肠的某些亚细胞颗粒中得到证实（TERENIUS 1972 和未发表的结果）。类似的方法现已应用于大鼠大脑皮层的各种亚细胞部分。这些实验证明 DHM 与突触质膜 (SPM) 部分（见下文）特异性结合，但不与其他部分（例如细胞核或线粒体）结合。 SPM级分的制备在0-4英寸下进行。将重125-150g的Sprague-Dawley品系的雌性大鼠放血，取出全脑并置于0.32M蔗糖中。分离大脑皮层并基本上按照GRAY.&WHITTAKER(1962)所述获得粗制线粒体级分。将粗制线粒体级分在30℃下进行渗透压休克和离心。 12,000 X g,,,,, 20 分钟。（WHITTAKER 等，1964）将上清液分层在由等体积的 0.4、1.0 和 1.2 M 蔗糖组成的不连续蔗糖梯度上，并离心 100,000 X g,,, 80 分钟。0.4 和 1.0 M 之间的条带。蔗糖被指定为SPM级分并取样用于培养。该级分的标记酶（乙酰胆碱酯酶和5'-核苷酸酶）的水平与预期一致，在4ml HEPES缓冲液（TF~RENIUS，未公开的结果）中振荡培养，预培养10分钟后。将DHM-7、8JH(比活性50mci/mg，放射化学纯度2·99yo)添加到0.2ml缓冲液中至终浓度0.6×10-9M并且
Knowledge of the subcellular localization of the receptor (s) for narcotic analgesics is important for understanding their mechanisms of action. A specific uptake of dihydromorphine-3H (DHM) has been demonstrated in certain subcellular particles of the guinea-pig ileum (TERENIUS 1972 and unpublished results). Similar methods have now been applied to various subcellular fractions of the rat cerebral cortex. These experiments demonstrated a specific binding of DHM to the synaptic plasma membrane (SPM) fraction (see below) but not to other fractions such as nuclei or mitochondria. The preparation of the SPM fraction was performed at 0-4". Female rats of the Sprague-Dawley strain weighing 125-150 g were bled and the whole brains were removed and placed in 0.32 M sucrose. The cerebral cortex was isolated and a crude mitochondrial fraction obtained essentially as described by GRAY. & WHITTAKER (1962). The crude mitochondrial fractions were subjected to osmotic shock and centrifugation at 12,000 X g,,,,, for 20 min.(WHITTAKER et ul. 1964). The supernatant was layered over a discontinuous sucrose gradient, consisting of equal volumes of 0.4, 1.0 and 1.2 M sucrose, and centrifuged for 100,000 X g,,, for 80 min. The band between 0.4 and 1.0 M sucrose was designated the SPM fraction and was sampled for incubations. Electron micrographs and levels of marker enzymes (acetylcholinesterase and 5'-nucleotidase) of this fraction were as expected. Aliquots of the SPM fraction were incubated at 25O with shaking in 4 mI of a HEPES buffer (TF~ RENIUS, unpublished results) which, when indicated contained non-labelled competitors. After pre-incubation for 10 min., DHM-7, 8JH (specific activity 50 mci/mg, radiochemical purity 2 99 yo) was added in 0.2 ml buffer to a final concentration of 0.6 X lO-9M and