背景：许多离子通道研究需要单个分散的神经元。人工原代培养的神经细胞受环境的影响，自身特性改变较大，而急性分离的神经元却能相对保持完好的生理特性。目的：建立急性分离尾核神经元的方法，用膜片钳技术研究尾核神经元离子通道及...背景：许多离子通道研究需要单个分散的神经元。人工原代培养的神经细胞受环境的影响，自身特性改变较大，而急性分离的神经元却能相对保持完好的生理特性。目的：建立急性分离尾核神经元的方法，用膜片钳技术研究尾核神经元离子通道及其信号转到机制，利于深入了解尾核的功能。设计、时间及地点：以细胞为对象的观察实验，于2008-02/12在三峡大学医学院实验中心完成。材料：新生7~10 d Wistar乳鼠12只，雌雄不限。方法：取7~10 d的大鼠，采用酶和机械分离法制备分散的单个尾核神经元，利用全细胞膜片钳技术记录电压依赖性钙电流。主要观察指标：用全细胞膜片钳急性分离大鼠尾核神经元的形态学视察和电生理特性。结果：用链蛋白酶（Protease）消化及机械分离法，急性分离的新生大鼠尾核神经元，表面光洁,胞膜完整，有较长的突起，形态和生理特性良好。利用急性分离的新生大鼠尾核神经元观察L-钙离子通道电流，所记录的电学参数在正常生理范围内，较好地保存电压依赖性钙离子通道活性。结论：分离出的神经元形态正常，有较长的轴突；保存了主要的离子通道活性，成功建立了一种适用于膜片钳技术的大鼠尾核神经元急性分离方法。展开更多
Objective: To study the influence of intra cerebroventricular injection of cholecystokinin octapeptide (CCK 8) on the effect of electroacupuncture (EA) i n antagonizing the electrical activity of pain related neurons ...Objective: To study the influence of intra cerebroventricular injection of cholecystokinin octapeptide (CCK 8) on the effect of electroacupuncture (EA) i n antagonizing the electrical activity of pain related neurons in caudate nucle us (Cd) and raising pain threshold.Methods: 30 male Wistar rats were used. Operations were performed under general anesthesia with 20% urethane (1.0 g/kg of body weight). The radiant heat irradiation (nociceptive stimulus) induce d rat tail flick reaction was used as the pain index. Extracellular discharges o f pain related neurons pain excitatory neurons (PEN) and pain inhibitory ne urons (PIN) in Cd and tail flick latency (TFL) before and after cerebroventri cular microinjection of CCK 8 (15 ng) were recorded. Electroacupuncture (EA) wa s applied to bilateral "Zusanli" (ST 36).Results: ① The radiant heat focused on the tail of rats caused increase of the pain evoked discharging frequency and shortening of the evoked discharging latency of PEN or reduction of the pain evoked discharging frequency and prolongation of the inhibitory du ration of the evoked discharges of PIN and generated tail flick reflex simultaneously. ② EA of bilateral "Zusanli" (ST 36) for 15 min resulted in inh ibition of the electric activity of PEN as well as potentiation of the electrica l activity of PIN and a prolongation of tail flick latency (TFL), i.e. exhibitin g the analgesic effect of EA. The effects peaked immediately after EA, the net i ncrease value (NIV) of the pain evoked discharges of 19 PENs was reduced from 1 6.17±2.30 Hz to 5.45±2.96 Hz and TFL was prolonged from 5.03±0.31 sec to 8.89 ±0.58 sec simultaneously, the inhibitory duration of the pain evoked discharge s of 12 PINs was shortened from 5.19±0.24 sec to 2.52±0.33 sec and TFL was pro longed from 4.57±0.23 sec to 8.12±0.29 sec simultaneously. These changes recov ered gradually 10 min after EA. ③ The inhibitory effect of EA on the pain evok ed discharges of PEN and the potentiated effect of EA on the electric activities of PIN, and the prolonged effect of EA on TFL were antagonized by intra cerebr oventricular injection of 15 ng CCK 8, i.e. CCK 8 could antagonize the analges ic effect of EA. About 4 min after injection of CCK 8, the effects were most ap parent. Very soon after EA, the NIV of 13 PENs was reduced from 9.36±2.10 Hz to 2.34±0.46 Hz and TFL was prolonged from 5. 38±0.18 sec to 8.60±0.49 sec simu ltaneously, the inhibitory duration of 10 PINs was shortened from 5.54±0.32 sec to 2.09±0.79 sec and TFL was prolonged from 4.92±0.17 sec to 9.44±0.21 sec s imultaneously. About 4 min after injection of CCK 8, the NIV of PENs was recov ered to 7.44± 1.38 Hz and TFL to 5.53±0.19 sec, the inhibitory duration of PINs was recovered to 6.20± 0.61 sec and TFL to 4.54±0.16 sec. About 14 min after injection of CCK 8, it gradually recovered. Conclusion: T he results demonstrate that the antagonism of CCK 8 on analgesic effect of EA s hows a coordinated and consistent action at the levels of electrical activity of central neurons and the whole behavior reaction.展开更多
文摘背景：许多离子通道研究需要单个分散的神经元。人工原代培养的神经细胞受环境的影响，自身特性改变较大，而急性分离的神经元却能相对保持完好的生理特性。目的：建立急性分离尾核神经元的方法，用膜片钳技术研究尾核神经元离子通道及其信号转到机制，利于深入了解尾核的功能。设计、时间及地点：以细胞为对象的观察实验，于2008-02/12在三峡大学医学院实验中心完成。材料：新生7~10 d Wistar乳鼠12只，雌雄不限。方法：取7~10 d的大鼠，采用酶和机械分离法制备分散的单个尾核神经元，利用全细胞膜片钳技术记录电压依赖性钙电流。主要观察指标：用全细胞膜片钳急性分离大鼠尾核神经元的形态学视察和电生理特性。结果：用链蛋白酶（Protease）消化及机械分离法，急性分离的新生大鼠尾核神经元，表面光洁,胞膜完整，有较长的突起，形态和生理特性良好。利用急性分离的新生大鼠尾核神经元观察L-钙离子通道电流，所记录的电学参数在正常生理范围内，较好地保存电压依赖性钙离子通道活性。结论：分离出的神经元形态正常，有较长的轴突；保存了主要的离子通道活性，成功建立了一种适用于膜片钳技术的大鼠尾核神经元急性分离方法。
文摘Objective: To study the influence of intra cerebroventricular injection of cholecystokinin octapeptide (CCK 8) on the effect of electroacupuncture (EA) i n antagonizing the electrical activity of pain related neurons in caudate nucle us (Cd) and raising pain threshold.Methods: 30 male Wistar rats were used. Operations were performed under general anesthesia with 20% urethane (1.0 g/kg of body weight). The radiant heat irradiation (nociceptive stimulus) induce d rat tail flick reaction was used as the pain index. Extracellular discharges o f pain related neurons pain excitatory neurons (PEN) and pain inhibitory ne urons (PIN) in Cd and tail flick latency (TFL) before and after cerebroventri cular microinjection of CCK 8 (15 ng) were recorded. Electroacupuncture (EA) wa s applied to bilateral "Zusanli" (ST 36).Results: ① The radiant heat focused on the tail of rats caused increase of the pain evoked discharging frequency and shortening of the evoked discharging latency of PEN or reduction of the pain evoked discharging frequency and prolongation of the inhibitory du ration of the evoked discharges of PIN and generated tail flick reflex simultaneously. ② EA of bilateral "Zusanli" (ST 36) for 15 min resulted in inh ibition of the electric activity of PEN as well as potentiation of the electrica l activity of PIN and a prolongation of tail flick latency (TFL), i.e. exhibitin g the analgesic effect of EA. The effects peaked immediately after EA, the net i ncrease value (NIV) of the pain evoked discharges of 19 PENs was reduced from 1 6.17±2.30 Hz to 5.45±2.96 Hz and TFL was prolonged from 5.03±0.31 sec to 8.89 ±0.58 sec simultaneously, the inhibitory duration of the pain evoked discharge s of 12 PINs was shortened from 5.19±0.24 sec to 2.52±0.33 sec and TFL was pro longed from 4.57±0.23 sec to 8.12±0.29 sec simultaneously. These changes recov ered gradually 10 min after EA. ③ The inhibitory effect of EA on the pain evok ed discharges of PEN and the potentiated effect of EA on the electric activities of PIN, and the prolonged effect of EA on TFL were antagonized by intra cerebr oventricular injection of 15 ng CCK 8, i.e. CCK 8 could antagonize the analges ic effect of EA. About 4 min after injection of CCK 8, the effects were most ap parent. Very soon after EA, the NIV of 13 PENs was reduced from 9.36±2.10 Hz to 2.34±0.46 Hz and TFL was prolonged from 5. 38±0.18 sec to 8.60±0.49 sec simu ltaneously, the inhibitory duration of 10 PINs was shortened from 5.54±0.32 sec to 2.09±0.79 sec and TFL was prolonged from 4.92±0.17 sec to 9.44±0.21 sec s imultaneously. About 4 min after injection of CCK 8, the NIV of PENs was recov ered to 7.44± 1.38 Hz and TFL to 5.53±0.19 sec, the inhibitory duration of PINs was recovered to 6.20± 0.61 sec and TFL to 4.54±0.16 sec. About 14 min after injection of CCK 8, it gradually recovered. Conclusion: T he results demonstrate that the antagonism of CCK 8 on analgesic effect of EA s hows a coordinated and consistent action at the levels of electrical activity of central neurons and the whole behavior reaction.
