中文摘要：

目的：从海马神经元谷氨酸离子型受体--AMPA受体亚基GluA1的831位丝氨酸（GluA1Ser831）磷酸化角度,探讨M1乙酰胆碱受体对AMPA受体GluA1亚基的调控作用及作用机制。方法：本研究以成熟的原代海马神经元为实验对象,用不易被降解的卡巴胆碱（Carbachol,CCh）作为胆碱受体激动剂,以免疫印迹法作为蛋白和磷酸化蛋白的主要检测手段,结合不同蛋白抑制剂研究M1受体调控AMPA受体GluA1亚基的关键信号分子及其机制。结果：1与对照组相比,CCh组Ser831的磷酸化水平显著升高。2CCh促进Ser831磷酸化的现象在M1受体选择性拮抗剂哌仑西平（Pirenzepine）＋CCh组消失,CCh升高GluA1-Ser831磷酸化水平的作用由M1受体介导。3蛋白激酶C（ProteinkinaseC,PKC）抑制剂白屈菜红碱（Chelerythrinechloride,CHCL）能对抗CCh促进GluA1-Ser831位点磷酸化的作用,而钙/钙调素依赖性蛋白激酶II（Calcium/calmodulin-dependentkinaseII,CaMKII）抑制剂KN62不能对抗CCh的作用。4为检测体内GluA1-Ser831的磷酸化情况,用小鼠海马组织定位注射CCh和CHCL,CCh组小鼠海马组织GluA1-Ser831位点的磷酸化水平升高,CHCL能对抗这种作用,PKC介导了M1受体激活所导致的GluA1-Ser831磷酸化水平的升高。结论：M1受体通过激活PKC促进GluA1-Ser831的磷酸化。

英文摘要：

Objective： By monitoring the phosphorylation of GluAl-Ser831 in hippocampal neurons, we explored the underlying mechanism of modulation of AMPA receptor by M1 muscarinic acetylcholine receptor. Methods： This study made use of the maturely cultured primary hippoeampal neurons and carbachol （CCh）, an acetylcholine receptor agonist which was resistant to degradation. West- ern Blot was used as the main method for the detection of protein and protein phosphorylation, and a variety of different protein inhibitors were utilized to study the signaling molecules of Ml receptor regulating phosphorylation ofGluA1 subunit. Results： ①CCh increased the phosphorylation of GluAl at Ser831 compared with control group. ②The up-regulated phosphorylation of GluA1-Ser831 by CCh was at- tenuated by added Pirenzepine in CCh, suggesting that CCh acted through M1 receptor to increase phosphorylation of GluA1-Ser831. ③ PKC inhibitor CHCL totally blocked the phosphorylation of Ser831 induced by CCh, whereas CAMKII inhibitor KN62 did not block the effects of CCh. ④ The hippocampus of mice were injected with CCh or CHCL to examine the effects in vivo. CCh increased the phos- phorylation of GIuA1 at Ser831. CHCL totally attenuated the increase of GluA1-Ser831 phosphorylation induced by CCh. Conclusion： The phosphorylation of GluA1 at Ser831 was promoted by M1 receptor through PKC.