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目的:研究P2X3受体(P2X3 receptor,P2X3R)在海马CA1区的定位及对CA1区锥体细胞兴奋性的影响。方法:免疫荧光技术对P2X3R在海马CA1区表达进行定位,SD鼠麻醉后断头取脑,修块后应用振动切片机切出350μm脑片并用正常人工脑脊液26℃恒温孵育1 h,通过膜片钳全细胞记录技术,给予P2X3R选择性拮抗剂AF-353和非选择性激动剂α,β-Me ATP干预,记录锥体细胞动作电位发放频率的改变。结果:P2X3R在海马区主要表达于神经元上,在星形胶质细胞不表达。P2X3R拮抗剂AF-353减慢锥体细胞动作电位发放频率减小其兴奋性[(0.788±0.163)Hz vs.(0.352±0.079)Hz,t=7.32,P=0.002],α,β-Me ATP则加快动作电位发放频率[(0.715±0.186)Hz vs.(1.610±0.454)Hz,t=-4.97,P=0.008],增加兴奋性。结论 :P2X3R在海马区主要表达于神经元树突及胞质并且影响神经元兴奋性,可能参与了兴奋性增高有关疾病的发生发展过程。
Objective: To investigate the localization of P2X3 receptor (P2X3R) in hippocampal CA1 region and its effect on the excitability of pyramidal cells in CA1 area. Methods: The expression of P2X3R in CA1 region of hippocampus was detected by immunofluorescence technique. After anesthesia, SD rats were decapitated and brains were harvested. The slices were excised and excised with 350μm slices and incubated with normal artificial cerebrospinal fluid at 26 ℃ for 1 h. Clamp whole cell recording technique. The P2X3R selective antagonist AF-353 and the non-selective agonist α, β-Me ATP were intervened to record the change of the distribution frequency of the action potential of pyramidal cells. Results: P2X3R mainly expressed in neurons in hippocampus, but not in astrocytes. The P2X3R antagonist AF-353 slowed the release of action potential of pyramidal cells and decreased its excitability [(0.788 ± 0.163) Hz vs. (0.352 ± 0.079) Hz, t = 7.32, P = 0.002] ATP increased the frequency of action potential [(0.715 ± 0.186) Hz vs. (1.610 ± 0.454) Hz, t = -4.97, P = 0.008] and increased excitability. CONCLUSION: P2X3R is mainly expressed in the dendrites and cytoplasm of neurons in the hippocampus and affects the excitability of neurons, which may be involved in the development and progression of diseases with increased excitability.