姓名施美琪.pptVIP

名詞解釋: Dopamine- and cAMP- regulated phosphoryprotein of 32 kDa 由dopamine 及 cAMP 所活化之磷酸蛋白(DARPP-32) Projection neuronA neuron that links one neuropil to one or more others Dopamine neurontransmitter CB1 receptor (cannabinoid receptor 1) 存在: basal ganglia(由putamen、caudate nucleus 、 globus pallidus組成 ),調控 motor behavior 實驗方向 1.利用不同的藥劑來測試DARPP-32磷酸化的程度 2.將Thr34 Ala34及Thr75 Ala75,並測試catalepsy情形 3.將Dopamine D2 receptor及Adenosine A2 receptor做基因剔除,比較其缺失對phospho-Thr34-DARPP-32的影響 Dopamine Receptor dopamine D2 receptor 活化後 adenylate cyclase cAMP PKA DARPP-32磷酸化 Materials Method Animal: Wild-type mice DARPP-32 knock-out mice (DARPP-32+/+) (DARPP-32-/-) Wild-type mice Dopamine D2 receptor knock-out mice Wild-type mice A2A receptor knock-out mice Drug: CP55,940 CB1 receptor agonist SR141716A CB1 receptor antagonist AM404 A inhibition of anandamide reuptake KW6002 A2A receptor antagonist 以上藥劑溶在含8% tween80 的食鹽水中 Measurnment of catalepsy Determination of phosphorylated DARPP-32 將不同的藥劑施打在老鼠身上 並在不同作用時間內將老鼠的頭剪下來 立即放入液態氮中(6s) 將striata分離出來 溶在 750μl SDS(1%) 煮10min 取3μl出來做protein 定量 跑SDS(10%, load30μg) 做western blot Result Activation of CB1 receptors stimulates phosphorylation of DARPP-32 at Thr34 Examined the ability of endogenous cannabioids (anandamide) to regulate DARPP-32 phosphorylation use AM404 PhosphoThr34-DARPP-32 is involved in CB1 receptor-mediated catalepsy Intact dopamine D2 and adenosine A2A receptor transmission are regulated for full-cannabinoid