贾炜姣^1,2,代广斌^2,3,耿国帅^2,3,杨忠波²,汤明杰²,修鹏⁴,张峰¹,王化斌^2,5

• 1:内蒙古农业大学 生命科学学院，内蒙古 呼和浩特 010018
• 2:中国科学院重庆绿色智能技术研究院 重庆市高分辨三维动态成像检测工程技术研究中心，重庆 400714
• 3:吉林大学 仪器科学与电气工程学院，吉林 长春 130000
• 4:浙江大学 工程力学系，浙江 杭州 310027

摘要(Abstract)：

细胞的生物学、化学及物理学等变化与细胞的生理功能密切相关。膜片钳技术是一种通过检测全细胞、单个或几个离子通道的电信号来研究细胞生理功能的重要技术。论文首先介绍了膜片钳技术，包括膜片钳的基本工作原理、细胞电信号的检测及分析方法；接着总结了膜片钳技术在揭示细胞电生理性质的化学及生物学机制方面的最新进展；然后概述了细胞电生理研究中膜片钳技术与其他生物检测技术的结合应用情况；最后对膜片钳技术的现状进行了总结并对其发展趋势进行了展望。

关键词(KeyWords)： 膜片钳技术;细胞;电生理;离子通道

Abstract:

Keywords:

基金项目(Foundation): 国家科技部重点研发专项（2016YFC0101002）；中央级大型科学仪器设备专项（YXGYQ201700136）；国家自然科学基金（11604332，11574268）；中国科学院“西部青年学者（重点）”计划（R52A500Z10）；重庆市基础前沿项目（cstc2015jcyjA10057）；重庆市应用开发项目（Y500061LH）。

作者(Author): 贾炜姣^1,2,代广斌^2,3,耿国帅^2,3,杨忠波²,汤明杰²,修鹏⁴,张峰¹,王化斌^2,5

DOI: 10.3969/j.issn.1003-9015.2018.04.004

参考文献(References)：

• [1] Chen P H, Zhang W, Zhou J, et al. Development of planar patch clamp technology and its application in the analysis of cellular electrophysiology[J]. Progress in Natural Science, 2009, 19(2):153-160.
  [2] Neher E, Sakmann B. Single-chanel currents recorded from membrane of denervated frog muscle-fibers[J]. Nature, 1976, 260(5554):799-802.
  [3] Farre C, Stoelzle S, Haarmann C, et al. Automated ion channel screening:patch clamping made easy[J]. Expert Opinion Therapeutic Targets, 2007, 11(4):557-565.
  [4] Shieh C C. Automated high-throughput patch-clamp techniques[J]. Drug Discovery Today, 2004, 9(13):551-552.
  [5] Yue J F, Qiao G H, Liu N, et al. Novel KCNQ2 channel activators discovered using fluorescence-based and automated patch-clamp-based high-throughput screening techniques[J]. Acta Pharmacologica Sinica, 2016, 37(1):105-110.
  [6] Almers W, Neher E. The Ca signal from fura-2 loaded mast-cells depends strongly on the method of dye-loading[J]. FEBS Letters, 1985, 192(1):13-18.
  [7] LIN Yan-fei (林燕飞), OUYANG-Shou (欧阳守). Recent progress and aplication of pathch clamp techniques (膜片钳技术研究进展及其应用)[J]. Strait Pharmaceutical Journal (海峡药学), 2008, 20(9):8-11.
  [8] Zhang D. Patch clamp:a powerful technique for studying the mechanism of acupuncture[J/OL]. Journal of Evidence-Based Complementary & Alternative Medicine, 2012[2018-01-05]. http://dx.doi.org/10.1155/2012/534219.
  [9] Wilson J R, Clark R B, Banderali U, et al. Measurement of the membrane potential in small cells using patch clamp methods[J]. Channels, 2011, 5(6):530-537.
  [10] Janve V S, Hernandez C C, Verdier K M, et al. Epileptic encephalopathy de novo GABRB mutations impair gamma-aminobutyric acid type a receptor function[J]. Annals of Neurology, 2016, 79(5):806-825.
  [11] Sherman A J, Shrier A, Cooper E. Series resistance compensation for whole-cell patch-clamp studies using a membrane state estimator[J]. Biophysical Journal, 1999, 77(5):2590-2601.
  [12] Xiong J, Hu G, Qu A L. Automatic calibration of frequency compensation system in computer-controlled patch-clamp amplifier[J]. Journal of Computer Science, 2007, 3(9):765-772.
  [13] Machtens J P, Kovermann P, Fahlke C. Substrate-dependent gating of anion channels associated with excitatory amino acid transporter 4[J]. Journal of Biological Chemistry, 2011, 286(27):23780-23788.
  [14] Adrian R H, Costantin L L, Peachey L D. Radial spread of contraction in frog muscle fibres[J]. Journal of Physiology-London, 1969, 204(1):231-257.
  [15] Ursu D, Schuhmeier R P, Melzer W. Voltage-controlled Ca²⁺ release and entry flux in isolated adult muscle fibres of the mouse[J]. Journal of Physiology-London, 2005, 562(2):347-365.
  [16] Karmazinova M, Lacinova L. Measurement of cellular excitability by whole cell patch clamp technique[J]. Physiological Research, 2010, 59:S1-S7.
  [17] Jurkat-Rott K, Lehmann-Hom F. The patch clamp technique in ion channel research[J]. Current Pharmaceutical Biotechnology, 2004, 5(4):387-395.
  [18] Nikolaev Y A, Dosen P J, Laver D R, et al. Single mechanically-gated cation channel currents can trigger action potentials in neocortical and hippocampal pyramidal neurons[J]. Brain Research, 2015, 1608:1-13.
  [19] QIAO Xiao-yan(乔晓艳), WU Jin-zhi(吴晋芝), GENG Xiao-yong(耿晓勇), et al. Computer simulation of cunent restoration for ionic sengle-channel (细胞膜离子单通道电流重构的计算机仿真)[J]. Computer Engineering and Appalication (计算机工程与应用), 2011, 47(16):218-220.
  [20] Lee C S K, Tong B C K, Cheng C W H, et al. Characterization of two-pore channel 2 by nuclear membrane electrophysiology[J/OL]. Scientific Reports, 2016[2018-01-05]. https://doi.org/10.1038/srep20282.
  [21] LIU Zhen-wei (刘振伟). Pratical patch clamp techniques (实用膜片钳技术)[M]. Beijing (北京):Military Medical Science Press (军事医学科学出版社), 2006:90-119.
  [22] Neher E. Ion channels for communication between and within cells[J]. Science, 1992, 256(5056):498-502.
  [23] Ridone P, Nakayama Y, Martinac B, et al. Patch clamp characterization of the effect of cardiolipin on MscS of E-coli[J]. European Biophysics Journalwith Biophysics Letters, 2015, 44(7):567-576.
  [24] LOU Xue-lin (娄雪林), ZHOU Zhuan (周专), KANG Hua-huang (康华光). Single channel and whole cell recordings using patch clamptechnique (单通道和全细胞记录技术)[J]. Chinese Journal of Medical Instrumentation (中国医疗器械杂志), 2000, 24(4):221-226.
  [25] Malboubi M, Jiang K. Gigaseal formation in patch clamping with applications of nanotechnology[M]. Berlin, Heidelberg:Springer, 2014:3-7.
  [26] Wu M J, Kee K H, Na J, et al. Pituitary adenylate cyclase-activating polypeptide inhibits pacemaker activity of colonic interstitial cells of cajal[J]. Korean Journal of Physiology and Pharmacology, 2015, 19(5):435-440.
  [27] Lolicato M, Arrigoni C, Mori T, et al. K2P2.1(TREK-1)-activator complexes reveal a cryptic selectivity filter binding site[J]. Nature, 2017, 547(7663):364-368.
  [28] Hamilton N B, Kolodziejczyk K, Kougioumtzidou E, et al. Proton-gated Ca²⁺-permeable TRP channels damage myelin in conditions mimicking ischaemia[J]. Nature, 2016, 529(7587):523-527.
  [29] Li M F, Toombes G E S, Silberberg S D, et al. Physical basis of apparent pore dilation of ATP-activated P2X receptor channels[J]. Nature Neuroscience, 2015, 18(11):1577-1583.
  [30] Azimi I, Roberts-Thomson S J, Monteith G R. Calcium influx pathways in breast cancer:opportunities for pharmacological intervention[J]. British Journal of Pharmacology, 2014, 171(4):945-960.
  [31] Berzingi S, Newman M, Yu H G. Altering bioelectricity on inhibition of human breast cancer cells[J/OL]. Cancer Cell International, 2016[2018-01-05]. https://doi.org/10.1186/s12935-016-0348-8.
  [32] Oehler B, Kistner K, Martin C, et al. Inflammatory pain control by blocking oxidized phospholipid-mediated TRP channel activation[J/OL]. Scientific Reports, 2017[2018-01-05]. https://doi.org/10.1038/s41598-017-05348-3.
  [33] Kheradpezhouh E, Ma L L, Morphett A, et al. TRPM2 channels mediate acetaminophen-induced liver damage[J]. Proceeding of the National Academy Sciences of the United States of America, 2014, 111(8):3176-3181.
  [34] Kheradpezhouh E, Panjehshahin M R, Miri R, et al. Curcumin protects rats against acetaminophen-induced hepatorenal damages and shows synergistic activity with N-acetyl cysteine[J]. European Journal of Pharmacology, 2010, 628(1-3):274-281.
  [35] Kheradpezhouh E, Barritt G J, Rychkov G Y. Curcumin inhibits activation of TRPM2 channels in rat hepatocytes[J]. Redox Biology, 2016, 7:1-7.
  [36] GE Man-ling (葛曼玲), ZHAO Quan-ming (赵全明), GUO Hong-yong (郭鸿涌), et al. The present patch-clamp technique in neuron electrical signal test (膜片钳技术在神经元电信号测试中的进展)[J]. Chinese Journal of Scientific Instrument (仪器仪表学报), 2002, 23(3):70-71.
  [37] Lu H C, Mackie K. An introduction to the endogenous cannabinoid system[J]. Biological Psychiatry, 2016, 79(7):516-525.
  [38] Gantz S C, Bean B P. Cell-autonomous excitation of midbrain dopamine neurons by endocannabinoid-dependent lipid signaling[J]. Neuron, 2017, 93(6):1375-1387.
  [39] Zeilhofer H U, Wildner H, Yevenes G E. Fast synaptic inhibition in spinal sensory processing and pain control[J]. Physiological Reviews, 2012, 92(1):193-235.
  [40] Lynch J W, Callister R J. Glycine receptors:A new therapeutic target in pain pathways[J]. Current Opinion in Investigational Drugs, 2006, 7(1):48-53.
  [41] Acuna M A, Yevenes G E, Ralvenius W T, et al. Phosphorylation state-dependent modulation of spinal glycine receptors alleviates inflammatory pain[J]. Journal of Clinical Investigation, 2016, 126(7):2547-2560.
  [42] Xiong W, Cui T X, Cheng K J, et al. Cannabinoids suppress inflammatory and neuropathic pain by targeting alpha 3 glycine receptors[J]. The Journal of Experimental Medicine, 2012, 209(6):1121-1134.
  [43] Islam R, Zhang Y, Xu L, et al. A chemogeneticreceptor that enhances the magnitude and frequency of glycinergic inhibitory postsynaptic currents without inducing a tonic chloride flux[J]. ACS Chemical Neuroscience, 2017, 8(3):460-467.
  [44] Yun I A, Wakabayashi K T, Fields H L, et al. The ventral tegmental area is required for the behavioral and nucleus accumbens neuronal firing responses to incentive cues[J]. Journal of Neuroscience, 2004, 24(12):2923-2933.
  [45] Zhang S L, Qi J, Li X P, et al. Dopaminergic and glutamatergic microdomains in a subset of rodent mesoaccumbens axons[J]. Nature Neuroscience, 2015, 18(3):386-392.
  [46] Berrios J, Stamatakis A M, Kantak P A, et al. Loss of UBE3A from TH-expressing neurons suppresses GABA co-release and enhances VTA-NAc optical self-stimulation[J/OL]. Nature Communications, 2016[2018-01-05]. https://doi.org/10.1038/ncomms10702.
  [47] Ossola D, Amarouch M Y, Behr P, et al. Force-controlled patch clamp of beating cardiac cells[J]. Nano Letters, 2015, 15(3):1743-1750.
  [48] Zhang C L, Li P, Liu L Q, et al. Development of mechanostimulated patch-clamp system for cellular physiological study[J]. IEEE/ASME Transactions on Mechatronics, 2014, 19(4):1138-1147.
  [49] Fertig N, Blick R H, Behrends J C. Whole cell patch clamp recording performed on a planar glass chip[J]. Biophysical Journal, 2002, 82(6):3056-3062.
  [50] Schmidt C, Mayer M, Vogel H. A chip-based biosensor for the functional analysis of single ion channels[J]. Angewandte Chemie International Edition, 2000, 39(17):3137-3140.
  [51] SHEN Ya-feng (沈亚峰), LIANG Wen (梁文), LEI Chang-hai (雷长海), et al. Synchronous observation of Ca²⁺ release in cardiac myocytes by patch clamp and confocal microscope (利用膜片钳-激光扫描共聚焦显微镜同步实时系统观察心肌细胞浆内钙离子的释放)[J]. Journal of Chinese Electron Microscopy Society (电子显微镜学报), 2008, 27(2):127-130.
  [52] Zheng J, Zagotta W N. Gating rearrangements in cyclic nucleotide-gated channels revealed by patch-clamp fluorometry[J]. Neuron, 2000, 28(2):369-374.
  [53] Kusch J, Biskup C, Thon S, et al. Interdependence of receptor activation and ligand binding in HCN2 pacemaker channels[J]. Neuron, 2010, 67(1):75-85.
  [54] Mony L, Berger T K, Isacoff E Y. A specialized molecular motion opens the Hv1 voltage-gated proton channel[J]. Nature Structural & Molecular Biology, 2015, 22(4):283-290.
  [55] CHENG Wei (程为), LI Mei (李玫), DU Sha (杜莎), et al. Development and application of patch-clamp fluorometryin life scienceresearch (荧光膜片钳新技术的应用及进展)[J]. Progress in Biochemistry and Biophysics (生物化学与生物物理进展), 2016, 43(2):187-192.
  [56] Ebina T, Sohya K, Imayoshi I, et al. 3D clustering of GABAergic neurons enhances inhibitory actions on excitatory neurons in the mouse visual cortex[J]. Cell Reports, 2014, 9(5):1896-1907.
  [57] Tao C, Zhang G W, Xiong Y, et al. Functional dissection of synaptic circuits:in vivo patch-clamp recording in neuroscience[J/OL]. Frontiers in Neural Circuits, 2015[2018-01-05]. https://doi.org/10.3389/fncir.2015.00023.