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An optimized recording method to characterize biophysical and pharmacological properties of acid-sensing ion channel
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作者 李爱 司文 +2 位作者 胡新武 刘长金 曹晓华 《Neuroscience Bulletin》 SCIE CAS CSCD 2008年第3期160-165,共6页
Objective To re-confirm and characterize the biophysical and pharmacological properties of endogenously expressed human acid-sensing ion channel 1a (hASIC1a) current in HEK293 cells with a modified perfusion methods... Objective To re-confirm and characterize the biophysical and pharmacological properties of endogenously expressed human acid-sensing ion channel 1a (hASIC1a) current in HEK293 cells with a modified perfusion methods. Methods With cell floating method, which is separating the cultured cell from coverslip and putting the cell in front of perfusion tubing, whole cell patch clamp technique was used to record hASICla currents evoked by low pH external solution. Results Using cell floating method, the amplitude of hASICla currents activated by pH 5.0 in HEK293 cells is twice as large as that by the conventional method where the cells remain attached to coverslip. The time to reach peak at two different recording conditions is (21±5) ms and (270±25) ms, respectively. Inactivation time constants are (496±23) ms and (2284±120) ms, respectively. The cell floating method significantly increases the amiloride potency of block on hASIC 1 a [IC50 is (3.4± 1.1 ) μmol/L and (2.4± 0.9) μmol/L, respectively]. Both recording methods have similar pH activation ECs0 (6.6±0.6, 6.6±0.7, respectively). Conclusion ASICs channel activation requires fast exchange of extracellular solution with the different pH values. With cell floating method, the presence of hASIC la current was re-confirmed and the biophysical and pharmacological properties of hASIC la channel in HEK293 cells was precisely characterized. This method could be used to study all ASICs and other ligand-gated channels that require fast extracellular solution exchange. 展开更多
关键词 acid-sensing ion channel patch-clamp recording PH
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Action Mechanism Research of Lanthanons to Slow Vacuolar Ion Channels in Raphanus Satirus L.(Xinlimei) Radish by Patch-Clamp 被引量:1
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作者 Pin YANG Yan Fang SHANG Zhen Ming PEI 《Chinese Chemical Letters》 SCIE CAS CSCD 2001年第9期813-814,共2页
We used whole-vacuolar patch-clamp recording mode to study the action mechanism of La3+ to Slow Vacuolar (SV) channels for the first time. We recorded SV channel currents of Xinlimei (Raphanus satirus L.) vacuolars. T... We used whole-vacuolar patch-clamp recording mode to study the action mechanism of La3+ to Slow Vacuolar (SV) channels for the first time. We recorded SV channel currents of Xinlimei (Raphanus satirus L.) vacuolars. The minimum activation potentials of voltage-dependent SV channels tied in 25+/-5 mV. The increase in cytoplasmic Ca2+ led to enhancement of SV-type currents. It was found that the threshold potential of activation shifted towards more depolarized values whenever cytoplasmic Ca2+ was increased. When 10(-10) mol/L free La3+ was added to the bath, SV-type current was suppressed by 60 similar to 75%. These data showed La3+ reduced ion permeabilities of Xinlimei root vacuolar membrane. 展开更多
关键词 Whole-vacuolar recording SV-type current cytoplasmic Ca2+ La3+ patch-clamp
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单通道和全细胞记录技术 被引量:11
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作者 娄雪林 周专 康华光 《中国医疗器械杂志》 CAS 2000年第4期221-226,205,共7页
离子通道的活动是可兴奋细胞传递信息的基础 ,膜片钳技术的发明导致了生命科学的一场革命。对膜片钳的工作模式、记录模式。
关键词 离子通道 膜片钳 单通道记录 全细胞记录
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5-HT增强大鼠骶髓后连合核神经元牛磺酸激活的全细胞电流 被引量:5
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作者 王殿仕 徐天乐 +1 位作者 庞志平 李继硕 《生理学报》 CAS CSCD 北大核心 1998年第6期656-662,共7页
应用制霉菌素穿孔全细胞记录方法,研究了5-羟色胺对急性分离的大鼠髓后连合核神经元磺酸激活的全细胞电流的调控。
关键词 5-羟色胺 牛磺酸 骶髓后连合核 电流
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穿支皮瓣床旁护理记录单的设计与应用 被引量:3
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作者 徐晓燕 钟玲 +4 位作者 董凯旋 李霞 徐兰 白艳 周智 《护理管理杂志》 2015年第7期526-527,共2页
目的探讨穿支皮瓣术后患者床旁护理记录单的使用效果。方法将穿支皮瓣床旁护理记录单应用于108例穿支皮瓣术后患者,观察使用记录单的书写质量、书写时间及患者、医生的满意度。结果术后发现动脉危象4例,静脉危象8例,皮瓣感染2例,经及时... 目的探讨穿支皮瓣术后患者床旁护理记录单的使用效果。方法将穿支皮瓣床旁护理记录单应用于108例穿支皮瓣术后患者,观察使用记录单的书写质量、书写时间及患者、医生的满意度。结果术后发现动脉危象4例,静脉危象8例,皮瓣感染2例,经及时对症治疗后皮瓣均存活;记录单书写合格率为97.22%,护士单次书写时间由(4.70±0.62)min缩短到1.51 min(P<0.01),患者对护理服务满意度为98.56%,医生满意度为96.37%。结论床旁护理记录单记录方法科学,不仅能客观、及时地反映皮瓣的动态信息,还能提高护士的工作效率。 展开更多
关键词 护理记录单 护理 穿支皮瓣
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用β-escin穿孔膜片技术研究粉防己碱对豚鼠心室肌钙电流的作用 被引量:1
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作者 傅丽英 李泱 +3 位作者 曾玉杰 姚伟星 夏国瑾 江明性 《药学学报》 CAS CSCD 北大核心 2002年第11期853-857,共5页
目的 用β escin穿孔膜片 (PPR)技术研究粉防己碱 (tetrandrine ,Tet)对ICa ,L 和ICa ,T的作用。方法 用PPR和全细胞记录 (WCR)模式记录心室肌细胞ICa ,L和ICa,T。结果  2 5 μmol·L- 1 β escin可在心室肌细胞形成稳定的PPR模... 目的 用β escin穿孔膜片 (PPR)技术研究粉防己碱 (tetrandrine ,Tet)对ICa ,L 和ICa ,T的作用。方法 用PPR和全细胞记录 (WCR)模式记录心室肌细胞ICa ,L和ICa,T。结果  2 5 μmol·L- 1 β escin可在心室肌细胞形成稳定的PPR模式 ,用此模式记录的ICa ,L衰减明显减慢。在PPR模式下 1~ 30 0 μmol·L- 1 Tet浓度依赖性地减小ICa ,L幅值。 3,30和30 0 μmol·L- 1 Tet对ICa ,T的抑制率分别为 (16± 5 ) % ,(40± 7) %和 (75± 11) %。结论 用 2 5 μmol·L- 1 β escin在豚鼠心室肌细胞能得到较稳定的PPR模式 ,在此模式下Tet浓度依赖性地抑制ICa ,L和ICa ,T。 展开更多
关键词 β-escin 穿孔膜片技术 粉防己碱 豚鼠 心室肌钙电流
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5-羟色胺增强甘氨酸激活的大鼠脊髓背角浅层神经元的全细胞电流 被引量:6
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作者 康杰芳 庞志平 李云庆 《神经解剖学杂志》 CSCD 北大核心 1998年第4期333-339,共7页
应用制霉菌素穿孔膜片钳全细胞记录技术研究了5-HT对急性分离的大鼠脊髓背角浅层(Ⅰ、Ⅱ层)神经元甘氨酸激活的全细胞电流的调控作用。实验结果表明:(1)在脊髓背角浅层神经元,甘氨酸作用于士的宁敏感型的甘氨酸受体,在钳制电位... 应用制霉菌素穿孔膜片钳全细胞记录技术研究了5-HT对急性分离的大鼠脊髓背角浅层(Ⅰ、Ⅱ层)神经元甘氨酸激活的全细胞电流的调控作用。实验结果表明:(1)在脊髓背角浅层神经元,甘氨酸作用于士的宁敏感型的甘氨酸受体,在钳制电位为-40mV时,可引起内向电流(IGly);(2)5-HT呈浓度依赖性的增强Gly反应;(3)5-HT的增强作用既不改变IGly的平衡电位,也不影响Gly与受体的亲和力;(4)5-HT2受体激动剂α-甲基-5-HT能模拟5-HT的增强效应;而5-HT2受体的拮抗剂ketanserine可阻断5-HT对IGly的增强作用;(5)蛋白激酶C(PKC)的抑制剂chelerythrine可抑制5-HT对IGly的增强作用。本研究的结果说明:5-HT可呈浓度依赖性地增强大鼠脊髓背角浅层神经元的IGly,5-HT经5-HT2受体介导其对IGly的调控,脊髓背角浅层神经元内经PKC的信号转导途径在5-HT对IGly的增强效应中起重要的作用。本研究的结果提示5-HT和Gly的相互作用在脊髓水平的伤害性信息传递和调控过程中可能具有重要的意义。 展开更多
关键词 5-HT 甘氨酸 脊髓背角浅层 大鼠 全细胞电流
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应用穿孔全细胞膜片钳记录技术研究培养大鼠耳蜗螺旋神经节神经元的基本电生理特性 被引量:1
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作者 查定军 王智明 +3 位作者 薛涛 林颖 邱建华 李云庆 《神经解剖学杂志》 CAS CSCD 北大核心 2008年第2期157-160,共4页
为了深入了解耳蜗螺旋神经节(SG)神经元的基本电生理特点,本研究首先成功培养了急性分离的大鼠SG神经元,然后采用穿孔全细胞膜片钳记录技术观察了培养SG神经元的基本电生理特性。结果表明,可从培养的SG神经元记录到动作电位,且不同的SG... 为了深入了解耳蜗螺旋神经节(SG)神经元的基本电生理特点,本研究首先成功培养了急性分离的大鼠SG神经元,然后采用穿孔全细胞膜片钳记录技术观察了培养SG神经元的基本电生理特性。结果表明,可从培养的SG神经元记录到动作电位,且不同的SG神经元具有异质性的放电特征。上述结果提示穿孔全细胞膜片钳记录技术可用于体外培养耳蜗SG神经元的电生理特征研究,为开展听觉信息传导和调控的功能研究开辟了新途径。 展开更多
关键词 螺旋神经节神经元 细胞培养 穿孔全细胞膜片钳记录 大鼠
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锌离子对急性分离的大鼠骶髓后连合核神经元GABA_A受体介导电流的抑制作用(英文) 被引量:1
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作者 庞志平 王殿仕 +1 位作者 徐天乐 李继硕 《生理学报》 CAS CSCD 北大核心 1998年第6期649-655,共7页
采用制霉菌素穿孔膜片箝技术,研究了锌离子(Zn2+)对急性分离的大鼠骶髓后连合核神经元GABAA受体介导电流的作用。结果表明:(1)在箝制电压为-40mV时,GABA可通过GABAA受体介导产生内向电流;(2)此电流可被Zn2+呈非电压依赖性可... 采用制霉菌素穿孔膜片箝技术,研究了锌离子(Zn2+)对急性分离的大鼠骶髓后连合核神经元GABAA受体介导电流的作用。结果表明:(1)在箝制电压为-40mV时,GABA可通过GABAA受体介导产生内向电流;(2)此电流可被Zn2+呈非电压依赖性可逆地阻断;(3)在Zn2+存在的情况下,GABA的浓度-效应曲线平行右移。上述结果提示,Zn2+可能通过变构调控机制对GANAA介导的反应产生抑制作用。 展开更多
关键词 ZN^2+ GABAA受体 骶髓后连合核 电流 神经元
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依托咪酯降低新生大鼠离体脊髓腹角神经元的兴奋性及抑制nAChR的功能 被引量:5
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作者 朱苏月 黄艳 +5 位作者 靳娜 杨鑫宇 张环环 徐爱萍 汪萌芽 郑超 《南方医科大学学报》 CAS CSCD 北大核心 2020年第5期676-682,共7页
目的研究依托咪酯(ET)对脊髓腹角神经元电生理特性及烟碱型乙酰胆碱受体(nAChR)的影响。方法选用19只7~12 d新生SD大鼠,麻醉后,将含有腰骶膨大的脊髓分离并切片,用木瓜蛋白酶(0.18 g/30 mL人工脑脊液)消化切片并孵育40 min,显微镜下选... 目的研究依托咪酯(ET)对脊髓腹角神经元电生理特性及烟碱型乙酰胆碱受体(nAChR)的影响。方法选用19只7~12 d新生SD大鼠,麻醉后,将含有腰骶膨大的脊髓分离并切片,用木瓜蛋白酶(0.18 g/30 mL人工脑脊液)消化切片并孵育40 min,显微镜下选取腹角,用抛光的巴斯德吸管进行急性机械分离神经元,对贴壁的健康神经元结合药理学方法进行穿孔膜片钳记录实验。在电流钳模式下,先记录脊髓腹角神经元自发动作电位(AP),然后结合预处理给药方式,分别记录不同浓度的ET对脊髓腹角神经元自发AP影响。在电压钳模式下,先应用尼古丁在脊髓腹角神经元诱导出内向电流,然后结合预处理给药方式,记录在不同浓度的ET、不同钳制电位以及不同使用时间的情况下,ET对尼古丁在脊髓腹角神经元诱导的内向电流的影响。结果急性分离的脊髓腹角神经元状态良好,具有形状多样的胞体和完整的突起;共记录到21例脊髓腹角神经元有自发AP,经0.3、3.0、30.0μmol/L(3.0μmol/L相当于临床浓度)不同浓度的ET持续灌流2 min后,与给药前比较,结果12例神经元的AP幅度、锋电位幅度及超射分别被浓度依赖性抑制(P<0.01);自发放电频率降低(P<0.01)。另外9例神经元的AP被3.0或30.0μmol/L的ET完全取消;在相同钳制电位下(VH=-70 mV),分别经0.3、3.0和30.0μmol/L的ET预处理2 min后,对0.4 mmol/L尼古丁诱导的电流幅度显示出浓度依赖性压抑作用(P<0.01,n=7)。将钳制电位分别设定为-30、-50、-70 mV,应用30.0μmol/L的ET预处理2 min后,对0.4 mmol/L尼古丁诱导的电流幅度呈电压依赖性压抑作用(P<0.01,每个钳制电位下n=6)。在30.0μmol/L的ET预处理6 min过程中,分别于0、2、4、6 min时先后4次暴露于0.4 mmol/L的尼古丁(每次暴露时间为2 s),随着暴露次数增多,尼古丁电流幅度逐渐减小;但若在6 min的预处理过程中,仅在开始(0 min)和结束(6 min)时两次暴露于相同浓度尼古丁,则6 min时,电流幅度抑制率较4次暴露尼古丁时的抑制率明显降低(P<0.01,n=6)。结论ET以浓度依赖的方式降低脊髓腹角神经元的兴奋性,并且以浓度依赖、电压依赖和使用依赖的方式压抑nAChR功能。 展开更多
关键词 依托咪酯 脊髓腹角神经元 N型乙酰胆碱受体 穿孔膜片钳记录
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Orexin-A对脊髓腹角神经元基本电生理参数及尼古丁电流的作用 被引量:1
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作者 黄艳 高凌云 +3 位作者 朱苏月 张环环 汪萌芽 郑超 《皖南医学院学报》 CAS 2019年第3期205-210,共6页
目的:研究orexin-A对脊髓腹角神经元基本电生理参数及烟碱型乙酰胆碱受体(nAChR)的作用。方法:使用7~12d的新生SD大鼠。麻醉后,将含有腰骶膨大的脊髓分离并切片。用木瓜蛋白酶(papain,0.18g/30mL人工脑脊液消化切片并孵育40min。显微镜... 目的:研究orexin-A对脊髓腹角神经元基本电生理参数及烟碱型乙酰胆碱受体(nAChR)的作用。方法:使用7~12d的新生SD大鼠。麻醉后,将含有腰骶膨大的脊髓分离并切片。用木瓜蛋白酶(papain,0.18g/30mL人工脑脊液消化切片并孵育40min。显微镜下选取腹角,使用抛光的巴斯德吸管进行神经元的急性机械分离。对贴壁的健康神经元进行穿孔膜片钳记录。结果:①急性分离的脊髓腹角神经元状态良好,具有形状多样的胞体和完整的突起;②9个急性分离的脊髓腹角神经元有自发动作电位;③持续灌流orexin-A(OXA)2min后,在5例细胞记录到4个脊髓腹角神经元的自发动作电位放电频率增加了(63.64±5.25)%,幅度等参数无明显变化;④在15个神经元,有12例细胞给予0.3mmol/L尼古丁诱导出内向电流,幅度为(142.97±75.02)pA,用100nmol/LOXA进行2min的预处理,可显著抑制电流幅度至(69.07±61.07)pA(P<0.001),抑制率为(54.75±22.62)%;⑤在9个神经元中有7例细胞给予尼古丁诱导的电流幅度为(129.31±69.38)pA,将100nmol/LOXA共同施用于神经元,尼古丁诱导的电流幅度降为(68.61±34.98)pA,在此基础上,通过施用orexin-1受体(OX1R)拮抗剂SB334867(10μmol/L)2min后可阻断OXA对尼古丁诱导电流的抑制作用,电流幅值为(93.46±56.45)pA。因此,SB334867可完全取消OXA对尼古丁电流的抑制作用。但对于另两例神经元,SB334867并未阻断OXA的抑制作用。结论:Orexin-A可能通过OX1R对脊髓大部分腹角神经元的尼古丁电流具有抑制作用,但不能排除orexin-2受体(OX2R)也参与OXA作用的可能性。 展开更多
关键词 OREXIN-A 脊髓腹角神经元 N型乙酰胆碱受体(nAChR) 穿孔膜片钳记录
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Effects of Calmodulin-dependent Protein Kinase Ⅱ Inhibitor,KN-93,on Electrophysiological Features of Rabbit Hypertrophic Cardiac Myocytes 被引量:2
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作者 柯俊 陈锋 +6 位作者 张存泰 肖幸 涂晶 戴木森 王晓萍 陈兵 陈敏 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2012年第4期485-489,共5页
Cardiac hypertrophy is an independent risk factor for sudden cardiac death in clinical settings and the incidence of sudden cardiac death and ventricular arrhythmias are closely related.The aim of this study was to de... Cardiac hypertrophy is an independent risk factor for sudden cardiac death in clinical settings and the incidence of sudden cardiac death and ventricular arrhythmias are closely related.The aim of this study was to determine the effects of the calmodulin-dependent protein kinase(CaMK) Ⅱ inhibitor,KN-93,on L-type calcium current(I Ca,L) and early after-depolarizations(EADs) in hypertrophic cardiomyocytes.A rabbit model of myocardial hypertrophy was constructed through abdominal aortic coarctation(LVH group).The control group(sham group) received a sham operation,in which the abdominal aortic was dissected but not coarcted.Eight weeks later,the degree of left ventricular hypertrophy(LVH) was evaluated using echocardiography.Individual cardiomyocyte was isolated through collagenase digestion.Action potentials(APs) and I Ca,L were recorded using the perforated patch clamp technique.APs were recorded under current clamp conditions and I Ca,L was recorded under voltage clamp conditions.The incidence of EADs and I ca,L in the hypertrophic cardiomyocytes were observed under the conditions of low potassium(2 mmol/L),low magnesium(0.25 mmol/L) Tyrode’s solution perfusion,and slow frequency(0.25-0.5 Hz) electrical stimulation.The incidence of EADs and I ca,L in the hypertrophic cardiomyocytes were also evaluated after treatment with different concentrations of KN-92(KN-92 group) and KN-93(KN-93 group).Eight weeks later,the model was successfully established.Under the conditions of low potassium,low magnesium Tyrode’s solution perfusion,and slow frequency electrical stimulation,the incidence of EADs was 0/12,11/12,10/12,and 5/12 in sham group,LVH group,KN-92 group(0.5 μmol/L),and KN-93 group(0.5 μmol/L),respectively.When the drug concentration was increased to 1 μmol/L in KN-92 group and KN-93 group,the incidence of EADs was 10/12 and 2/12,respectively.At 0 mV,the current density was 6.7±1.0 and 6.3±0.7 PA·PF-1 in LVH group and sham group,respectively(P>0.05,n=12).When the drug concentration was 0.5 μmol/L in KN-92 and KN-93 groups,the peak I Ca,L at 0 mV was decreased by(9.4±2.8)% and(10.5±3.0)% in the hypertrophic cardiomyocytes of the two groups,respectively(P>0.05,n=12).When the drug concentration was increased to 1 μmol/L,the peak I Ca,L values were lowered by(13.4±3.7)% and(40±4.9)%,respectively(P<0.01,n=12).KN-93,a specific inhibitor of CaMKII,can effectively inhibit the occurrence of EADs in hypertrophic cardiomyocytes partially by suppressing I Ca,L,which may be the main action mechanism of KN-93 antagonizing the occurrence of ventricular arrhythmias in hypertrophic myocardium. 展开更多
关键词 calmodulin-dependent protein kinase KN-93 myocardial hypertrophy ELECTROPHYSIOLOGY perforated patch recording techniques
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Effects of antigliomatin from the scorpion venom of Buthus martensii Karsch on chloride channels on C6 glioma cells 被引量:1
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作者 Zan Wang Mingxian Li +4 位作者 Hongmei Meng Min Huang Weihong Lin Li Cui Shao Wang 《Neural Regeneration Research》 SCIE CAS CSCD 2011年第18期1365-1369,共5页
Using whole-cell patch-clamp recordings, the effects of antigliomatin were observed on chloride channels on C6 glioma cells cultured in vitro. Antigliomatin was extracted from the venom of the scorpion Buthus martensi... Using whole-cell patch-clamp recordings, the effects of antigliomatin were observed on chloride channels on C6 glioma cells cultured in vitro. Antigliomatin was extracted from the venom of the scorpion Buthus martensii Karsch. Chloride channels are closed under normal osmotic pressure. When osmotic pressure was reduced to 120, 110 and 100 mV, the cell volume enlarged, chloride channels opened, and the chloride channel current increased. Three minutes after antigliomatin treatment, the chloride channel current decreased in a dose-dependent manner. These results show that antigliomatin extracted from the venom of the scorpion Buthus martensii Karsch diminishes chloride channel currents on C6 glioma cells. 展开更多
关键词 ANTIGLIOMATIN C6 glioma cells chloride channels osmotic pressure whole-cell patch-clamp recording
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A Persistent Na<sup>+</sup>Current and Its Contribution to Burst-Like Firing in Ventral Tegmental Area Dopamine Neurons
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作者 Susumu Koyama Munechika Enjoji +1 位作者 Mark S. Brodie Sarah B. Appel 《Journal of Biomedical Science and Engineering》 2015年第7期429-440,共12页
The ventral tegmental area dopamine (DA VTA) neurons have the spontaneous tonic activity and an alteration of firing pattern from tonic to burst accelerates dopamine transmission more effectively in the mesoaccumbal d... The ventral tegmental area dopamine (DA VTA) neurons have the spontaneous tonic activity and an alteration of firing pattern from tonic to burst accelerates dopamine transmission more effectively in the mesoaccumbal dopaminergic system, leading to the reinforcing process of drugs of abuse such as alcohol and nicotine. In the present study, we examined whether a persistent Na+ current would contribute to burst firing in DA VTA neuronsusing nystatin-perforated recording. Tetrodotoxin (TTX) (1 μM) or riluzole (10 μM) hyperpolarized the membrane potential and stopped spontaneous firing of DA VTA neurons. In voltage-clamp analysis, a TTX and riluzole-sensitive and persistent Na+ current was activated at ?60 mV and reached maximal amplitude at ?40 mV. This persistent Na+ current was potentiated by a negative shift of the voltage of activation by eliminating Ca2+ from the extracellular solution. The Ca2+-free extracellular solution depolarized the membrane potential and increased the firing frequency of DA VTA neurons. When a continuous hyperpolarizing current was injected, the firing pattern of the DA VTA neurons transformed into burst-like firing;with average spike number of 4.9, average inter-spike interval of 221 ms, and an average plateau potential, on which the train of spikes generated, was 11 mV. The burst-like firing of DA VTA neurons was abolished by 10 μM riluzole. The concurrent blockade of both T-type Ca2+ current and small conductance Ca2+-activated K+(SK) currents by 100 μM nickel did not induce burst-like firing with or without continuous hyperpolarizing current injection in DA VTA neurons. In conclusion, increases in a persistent Na+ current that mediates a depolarizing driving force by removing extracellular Ca2+ contributes to burst-like firing in DA VTA neurons. 展开更多
关键词 APAMIN Acutely Dissociated Neurons EXTRACELLULAR Ca2+ Nickel NYSTATIN perforated Patch recording
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Electrophysiology of Sodium Receptors in Taste Cells
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作者 Albertino Bigiani 《Journal of Biomedical Science and Engineering》 2016年第8期367-383,共17页
Sodium intake is important to maintain proper osmolarity and volume of extracellular fluid in vertebrates. The ability to find sources of sodium ions for managing electrolyte homeostasis relies on the activity of the ... Sodium intake is important to maintain proper osmolarity and volume of extracellular fluid in vertebrates. The ability to find sources of sodium ions for managing electrolyte homeostasis relies on the activity of the taste system to sense salt. Several studies have been performed to understand the mechanisms underlying Na+ reception in taste cells, the peripheral detectors for food chemicals. It is now generally accepted that Na+ interacts with specific ion channels in taste cell membrane, called sodium receptors. As ion channels, these proteins mediate transmembrane ion fluxes (that is, electrical currents) during their operation. Thus, a lot of information on the functional properties of sodium receptors has been obtained by using electrophysiological techniques. Here, I review our current knowledge on the biophysical and physiological features of these receptors obtained by applying the patch-clamp recording techniques to single taste cells. 展开更多
关键词 Sodium Taste Epithelial Sodium Channel patch-clamp recording
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The Effects of Zinc and Other Divalent Cations on M-Current in Ventral Tegmental Area Dopamine Neurons
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作者 Susumu Koyama Munechika Enjoji +1 位作者 Mark S. Brodie Sarah B. Appel 《Journal of Biomedical Science and Engineering》 2014年第14期1075-1087,共13页
Ventral tegmental area dopamine (DA VTA) neurons are important for the reinforcing effects of drugs of abuse such as ethanol and nicotine. We have previously shown that M-current (IM) regulates the excitability of DA ... Ventral tegmental area dopamine (DA VTA) neurons are important for the reinforcing effects of drugs of abuse such as ethanol and nicotine. We have previously shown that M-current (IM) regulates the excitability of DA VTA neurons. Zinc (Zn2+) contributes to the regulation of neuronal excitation as a neuromodulator. In the present study, we investigated zinc effect on the properties of IM and the spontaneous firing frequency of DA VTA neurons. The standard deactivation protocol was used to measure IM during voltage-clamp recording with a hyperpolarizing voltage step to ﹣40 mV from a holding potential (VH) of ﹣25 mV. Zn2+ (100 μM) inhibited IM amplitude and IM recovered completely from the inhibition after the washout of Zn2+. Zn2+ inhibited IM in a concentration-dependent manner (IC50: 5.8 μM). When hyperpolarizing voltage steps were given to ﹣65 mV (in 10 mV increments) from a VH of ﹣25 mV, Zn2+ (100 μM) reduced IM amplitude at each voltage and zinc inhibition of IM was not voltage-dependent. Zn2+ increased the spontaneous firing frequency of DA VTA neurons in a concentration-dependent manner, suggesting that Zn2+ causes excitation of DA VTA neurons through an action on IM. IM of DA VTA neurons was inhibited by 100 μM divalent cations in increasing order of potency: Ba2+ (16%) 2+ (25%) 2+ (40%) 2+ (59%) 2+ (67%). These results suggest that Zn2+ may exert physiologically significant regulation of neuronal excitability in DA VTA neurons. 展开更多
关键词 DIVALENT CATION DOPAMINERGIC Nystatin-perforated PATCH recording ZINC
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Axonal bleb recording 被引量:4
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作者 Wenqin Hu Yousheng Shu 《Neuroscience Bulletin》 SCIE CAS CSCD 2012年第4期342-350,共9页
Patch-clamp recording requires direct accessibility of the cell membrane to patch pipettes and allows the investigation of ion channel properties and functions in specific cellular compartments.The cell body and relat... Patch-clamp recording requires direct accessibility of the cell membrane to patch pipettes and allows the investigation of ion channel properties and functions in specific cellular compartments.The cell body and relatively thick dendrites are the most accessible compartments of a neuron,due to their large diameters and therefore great membrane surface areas.However,axons are normally inaccessible to patch pipettes because of their thin structure;thus studies of axon physiology have long been hampered by the lack of axon recording methods.Recently,a new method of patchclamp recording has been developed,enabling direct and tight-seal recording from cortical axons.These recordings are performed at the enlarged structure(axonal bleb) formed at the cut end of an axon after slicing procedures.This method has facilitated studies of the mechanisms underlying the generation and propagation of the main output signal,the action potential,and led to the finding that cortical neurons communicate not only in action potential-mediated digital mode but also in membrane potential-dependent analog mode. 展开更多
关键词 AXON patch-clamp recording ion channel action potential
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Study of transmembrane La^(3+) movement in rat ventricular myocytes by the patch-clamp technique 被引量:3
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作者 YANG Pin, DU Huizhi & XUE ShaowuInstitute of Molecular Science, Shanxi University, Taiyuan 030006, China 《Chinese Science Bulletin》 SCIE EI CAS 2002年第18期1518-1522,共5页
We have studied transmembrane La3+ movement in rat ventricular myocytes for the first time by using the whole-cell patch-clamp recording mode. La3+ (0.01-5.0 mmol/L) could not bring out inward currents through the L-t... We have studied transmembrane La3+ movement in rat ventricular myocytes for the first time by using the whole-cell patch-clamp recording mode. La3+ (0.01-5.0 mmol/L) could not bring out inward currents through the L-type calcium channel in rat ventricular myocytes, while it could enter the cells by the same way carried by 1μmo1/L ionomycin. When the outward Na+ concentration gradient is formed, La3+ can enter the cells via Na-Ca exchange, and the exchange currents increase with the increase of external La3+ concentrations. But compared with Na-Ca exchange currents in the same concentration, the former is only 14%-38% of the latter. The patch-clamp experiment indicates that La3+ normally can not enter ventricular myocytes through L-type calcium channel, but it can enter the cells via Na-Ca exchange. 展开更多
关键词 WHOLE-CELL patch-clamp recording ventricular MYOCYTE L-TYPE calcium channel Na-Ca exchange La3+ Ca2+.
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Effects of BaCl_2 on slow vacuolar ion channels on radish by patch-clamp
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作者 杨频 张丽平 《Science China Chemistry》 SCIE EI CAS 2003年第1期84-89,共6页
The effects of BaCl_2 on slow vacuolar (SV) currents of radish are studied byusing the whole-vacuolar patch-clamp recording mode. The Ca^(2+)-dependent SV channel can beactivated by cytosolic Ca^(2+). When 1 mmol/L Ba... The effects of BaCl_2 on slow vacuolar (SV) currents of radish are studied byusing the whole-vacuolar patch-clamp recording mode. The Ca^(2+)-dependent SV channel can beactivated by cytosolic Ca^(2+). When 1 mmol/L BaCI_2 is added into pipette solution, SV currents aresuppressed remarkably. Then adding BaCI_2 of different concentrations into the bath solution, SVcurrents reflect different effects. The results show that BaCl_2 with a lower concentration (< 3mmol/L) promotes the channel currents and the currents are saturated when BaCl_2 concentrations arebetween 1 μmol/L and 1 mmol/L, but BaCl_2 with higher concentration (≥ 3 mmol/L) inhibits SVcurrents. 展开更多
关键词 patch-clamp whole-vacuolar recording SLOW VACUOLAR channel CYTOSOLIC Ca2+ Ba2+.
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钙调蛋白激酶Ⅱ抑制剂对肥厚心肌细胞的影响 被引量:2
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作者 柯俊 陈锋 +5 位作者 肖幸 戴木森 王晓萍 陈兵 陈敏 张存泰 《中华急诊医学杂志》 CAS CSCD 北大核心 2012年第2期151-155,共5页
目的 观察钙调蛋白激酶Ⅱ(CaMKⅡ)抑制剂KN-93对肥厚心肌细胞L型钙电流(ICa,L)及细胞内钙离子浓度([Ca2+]i)的影响.方法 选取雌性新西兰大白兔48只,随机(随机数字法)分为4组:假手术组(sham组)、心肌肥厚组(LVH组)、心肌... 目的 观察钙调蛋白激酶Ⅱ(CaMKⅡ)抑制剂KN-93对肥厚心肌细胞L型钙电流(ICa,L)及细胞内钙离子浓度([Ca2+]i)的影响.方法 选取雌性新西兰大白兔48只,随机(随机数字法)分为4组:假手术组(sham组)、心肌肥厚组(LVH组)、心肌肥厚+KN-93组(KN-93组)、心肌肥厚+KN-92组(KN-92组),每组12只,通过缩窄腹主动脉制备兔心肌肥厚模型,Sham组仅游离腹主动脉未进行缩窄.8周后,采用胶原酶消化法分离单个心肌细胞,应用穿孔膜片钳技术记录L型钙电流(ICa,L);应用钙荧光指示剂Fura-2/AM结合图像分析技术测定各组心肌细胞内[Ca2+]i.结果 8周后,心肌肥厚模型建立成功.在0 mV时LVH组、Sham组的峰值ICa.L分另为(1.38±0.3)nA、(0.87±0.1)nA(P<0.01,n=12),电流密度分别为(6.7±1.0)pA/pF、(6.3 ±0.7)pA/pF(P>0.05,n=12).当KN-92及KN-93在浓度为0.5μmol/L时,可分别使肥厚心肌细胞0 mV时的峰值ICa,L降低(9.4±2.8)%、(10.5±3)%(P>0.05,n=12);当浓度增至1 μmol/L时,其峰值ICa,L降低程度分别为(13.4±3.7)%、(40±4.9)%(P<0.01,n=12).Sham组、LVH组、KN-92组及KN-93组中心肌细胞[Ca2+]i分别为(98.0±12.3)nmol/L、(154.0±26.2)nmol/L、(147.0±29.6)nmol/L和(108.0±21.2)nmol/L.结论 CaMKⅡ特异性抑制剂KN-93可有效抑制肥厚心肌细胞ICa.L,减轻细胞内钙超载,这可能是其抗肥厚心肌室性心律失常发生的主要细胞电生理机制. 展开更多
关键词 钙调蛋白激酶Ⅱ KN-93 心肌肥厚 心肌细胞 电生理 L型钙电流 细胞内钙 离子浓度 穿孔膜片钳技术
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