Autoradiographic localization of delta opioid receptors in the rat brain using a highly selective bis-penicillamine cyclic enkephalin analog.
Autoradiographic localization of delta opioid receptors in the rat brain using a highly selective bis-penicillamine cyclic enkephalin analog.
复制标题
使用高选择性双青霉胺环状脑啡肽类似物对大鼠大脑中 δ 阿片受体进行放射自显影定位。
DOI：
10.1016/0014-2999(85)90770-8
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发表时间：
1985
影响因子：
5
通讯作者：
Yamamura,HI
中科院分区：
文献类型：
--
作者：
Gulya,K;Gehlert,DR;Wamsley,JK;Mosberg,HI;Hruby,VJ;Duckles,SP;Yamamura,HI
Mosberg等人实施了一种设计对8种阿片受体具有选择性的配体的开创性方法。（1983），他证明了环状构象限制性脑啡肽类似物（2-D-青霉胺，5-D-青霉胺）脑啡肽（DPDPE）对8种阿片受体显示出极高的特异性。最近Akiyama et al.（1985）首次提供了氚标记的DPDPE与大鼠脑和神经母细胞瘤-神经胶质瘤杂交（NG 108-15）细胞中8种阿片受体结合的体外表征。在本研究中，我们研究了阿片受体在大鼠中枢神经系统中的光镜放射自显影分布，采用高选择性δ激动剂[3 H] DPDPE（SA= 40 Ci/mmol，阿默舍姆Corp，阿灵顿Heights，IL.）。通过断头处死雄性Sprague-Dawley大鼠（150-200 g），取出脑并用塑料包埋介质（OCT Compound，Lab-Tek Products）包被到切片机卡盘上，并通过浸入液氮中冷冻。切下矢状面和冠状面切片（20 μ m），解冻并固定在铬明矾/明胶包被的载玻片上，并在室温下风干。将载玻片固定的切片在含有5 mMMgCl 2、2 mg/ml BSA、20 μ g/ml杆菌肽和100 mMNaCl的50 mMTris-HCl缓冲液（pH7.4，25 ℃）中于25 ℃预孵育15分钟，以减少任何内源性阿片样物质
A pioneering approach to the design of a ligand selective for the 8 opioid receptors was implemented by Mosberg et al.(1983) who demonstrated that the cyclic conformationally restricted enkephalin analog (2-D-penicillamine, 5-D-penicillamine) enkephalin (DPDPE) displayed extremely high specificity toward the 8 opioid receptors. Recently Akiyama et al.(1985) provided the first in vitro characterization of tritium labeled DPDPE binding to 8 opioid receptors in the rat brain and neuroblastoma-glioma hybrid (NG 108-15) cells. In the present study we examined the light microscopic autoradiographic distribution of opioid receptors in the rat central nervous system employing the highly selective 8 agonist [3H] DPDPE (SA= 40 Ci/mmol, Amersham Corp, Arlington Heights, IL.). Male Sprague-Dawley rats (150-200 g) were killed by decapitation, and the brain was removed and coated with plastic embedding medium (OCT Compound, Lab-Tek Products) onto microtome chucks and frozen by immersion into liquid nitrogen. Saggital and coronal sections (20# m) were cut, thawed and mounted onto chromealum/gelatin coated glass slides and air dried at room temperature. Slide-mounted sections were preincubated for 15 min at 25 C in 50 mM Tris-HCI buffer (pH 7.4 at 25 C) containing 5 mM MgC12, 2 mg/ml BSA, 20 gg/ml bacitracin and 100 mM NaC1, to reduce any endogenous opioids