DETERMINATION OF ENDOGENOUS ACETYLCHOLINE-RELEASE IN FREELY MOVING RATS BY TRANSSTRIATAL DIALYSIS COUPLED TO A RADIOENZYMATIC ASSAY - EFFECT OF DRUGS
DETERMINATION OF ENDOGENOUS ACETYLCHOLINE-RELEASE IN FREELY MOVING RATS BY TRANSSTRIATAL DIALYSIS COUPLED TO A RADIOENZYMATIC ASSAY - EFFECT OF DRUGS
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DOI：
10.1111/j.1471-4159.1987.tb05686.x
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发表时间：
1987-05-01
影响因子：
4.7
通讯作者：
VEZZANI, A
中科院分区：
文献类型：
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作者：
CONSOLO, S;WU, CF;VEZZANI, A
脑内透析技术与敏感且特异的放射酶法相结合，用于从自由活动的大鼠纹状体中回收和定量内源性细胞外乙酰胆碱。将细透析管横向插入尾状核，并用pH 6.1的林格溶液以2μl min-1的恒定速率灌注该管。每隔 10 分钟收集一次灌注液。在1和10μM毒扁豆碱存在下，乙酰胆碱释放为4.5±。 0.02 和 7.3 .+-。分别为 0.3 pmol/10 分钟（未针对回收率进行校正）。所有实验中均使用后一浓度的乙酰胆碱酯酶抑制剂。在基础条件下，乙酰胆碱输出至少在 4 小时内保持稳定。去极化 K+ 浓度使乙酰胆碱输出急剧、可逆地增加 87%。基础和 K+ 刺激的释放均依赖于 Ca2+。胆碱摄取抑制剂 hemicholinium-3（20μg 脑室内）在 90 分钟内将纹状体乙酰胆碱输出减少至基础值的 35％。东莨菪碱 (0.34 mg/kg s.c.) 引起乙酰胆碱释放的急剧增强。比基础值减少 63%，而氧化震颤素 (0.53 mg/kg i.p.) 短暂地将乙酰胆碱释放减少 54%。这些结果表明经纹状体透析监测内源性乙酰胆碱释放的生理和药理学适用性。
The technique of intracerebral dialysis in combination with a sensitive and specific radioenzymatic method was used for recovery and quantification of endogenous extracellular acetylcholine from the striata of freely moving rats. A thin dialysis tube was inserted transversally through the caudate nuclei, and the tube was perfused with Ringer solution, pH 6.1, at a constant rate of 2 .mu.l min-1. The perfusates were collected at 10-min intervals. In the presence of 1 and 10 .mu.M physostigmine, acetylcholine release was 4.5 .+-. 0.02 and 7.3 .+-. 0.3 pmol/10 min, respectively (not corrected for recovery). The latter concentration of the acetylcholinesterase inhibitor was used in all experiments. Under basal conditions, acetylcholine output was stable over at least 4 h. A depolarizing K+ concentration produced a sharp, reversible 87% increase in acetylcholine output. Both the basal and K+-stimulated release were Ca2+ dependent. The choline uptake inhibitor hemicholinium-3 (20 .mu.g intracerebroventricularly) reduced striatal acetylcholine output to 35% of the basal value within 90 min. Scopolamine (0.34 mg/kg s.c.) provoked a sharp enhancement of acetylcholine release of .apprx. 63% over basal values, whereas oxotremorine (0.53 mg/kg i.p.) transiently reduced acetylcholine release by 54%. These results indicate the physiological and pharmacological suitability of transstriatal dialysis for monitoring endogenous acetylcholine release.