Halothane disruption of alpha 2-adrenergic receptor-mediated inhibition of adenylate cyclase and receptor G-protein coupling in rat brain.
Halothane disruption of alpha 2-adrenergic receptor-mediated inhibition of adenylate cyclase and receptor G-protein coupling in rat brain.
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氟烷破坏大鼠脑中 α2-肾上腺素能受体介导的腺苷酸环化酶和受体 G 蛋白偶联的抑制作用。
DOI：
10.1016/0006-2952(90)90672-8
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发表时间：
1990
影响因子：
5.8
通讯作者：
Aronstam,RS
中科院分区：
文献类型：
--
作者：
Baumgartner,MK;Dennison,RL;Narayanan,TK;Aronstam,RS
方法将雄性Wistar大鼠（125-150 g;哈兰Sprague-Dawley，Indianapolis，IN）断头，取出其大脑皮质并在10体积的50 mM Tris-HCl，nH 7.4中匀浆。含有2 mM M&I和1 mM二硫代噻吩（DD？）*使用一个？聚四氟乙烯玻璃组织研磨机。将匀浆在4”下以17，000 g离心20 min。将沉淀重悬，用原始缓冲液洗涤两次，按照Salomon等人的方法测定腺苷酸环化酶活性。[lo]做了一些小的改动每个测定管含有最终体积为250 μ L的以下试剂：25 mM HEPES，pH 7.5; 1.33 mM EGTA，1 mM DTT; 2 mM MgS 〇 4，100 mM NaCl; 0.5 mM 3-异丁基-1-甲基黄嘌呤; 0.1 mM ATP; 1 mM CAMP; 1 PM GTP; 20 mM磷酸肌酸; 10单位肌酸磷酸激酶; 0.5 mM [8 * P] ATP。和100-150 pg蛋白质。通过Mao和Guidotti [II]的方法进行CAMP的纯化。
MethodsMale Wistar rats (125-150 g; Harlan Sprague-Dawley, Indianapolis, IN) were decapitated, and their cerebral cortices w&e removed and homogenized in 10 vol. of 50 mM Tris-HCI, nH 7.4. containing 2 mM M&I, and 1 mM dithiothr&bl (DD?)* using a?‘eflon-glass tissue grinder. The homogenate was spun at 17,000 g for 20 min at 4”. The pellet was resuspended and washed twice with the original buffer.Adenylate cyclase activity was measured following the method of Salomon et al.[lo] with minor modifications. Each assay tube contained the following reagents in a final volume of 25OpL: 25 mM HEPES, pH 7.5; 1.33 mM EGTA, 1 mM DTT; 2 mM MgS04, lOO mM NaCI; 0.5 mM 3-isobutyl-1-methylxanthine; 0.1 mM ATP; 1 mM CAMP; 1 PM GTP; 20 mM creatine phosphate; 10 units of creatine phosphokinase; 0.5 &i [&* P] ATP; and 100-150 pg protein. Purification of CAMP was carried out by the method of Mao and Guidotti [ll].