Overview of Electrophysiological Techniques

Alan D. Wickenden1

1 Janssen R&D, San Diego, California
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 11.1
DOI:  10.1002/0471141755.ph1101s64
Online Posting Date:  March, 2014
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Abstract

This unit provides an overview of the principal electrophysiological techniques commonly used for the study of ionic currents and the ion channels that mediate them. These techniques include electroencephalograms (EEGs), electrocardiograms (ECGs), single‐ and multiunit extracellular recording, multielectrode arrays, transepithelial recording, impedance measurements, and current‐clamp, voltage‐clamp, patch‐clamp, and lipid bilayer recording. The unit also discusses recent advances in high‐throughput, automated electrophysiological techniques for drug discovery and the use of stem cells as a tissue source. Curr. Protoc. Pharmacol. 64:11.1.1‐11.1.17. © 2014 by John Wiley & Sons, Inc.

Keywords: extracellular recording; intracellular recording; patch clamp; planar patch clamp; drug discovery; stem cells

     
 
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Table of Contents

  • Introduction
  • Electrophysiological Techniques
  • Electrophysiology and Drug Discovery
  • Tissue Sources for Electrophysiology
  • Outsourcing
  • Acknowledgements
  • Literature Cited
  • Figures
     
 
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Materials

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Figures

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Literature Cited

Literature Cited
  Asai, Y., Tada, M., Otsuji, T.G., and Nakatsuji, N. 2010. Combination of functional cardiomyocytes derived from human stem cells and a highly‐efficient microelectrode array system: An ideal hybrid model assay for drug development. Curr. Stem Cell Res. Therapy 5:227‐232.
  Asmild, M., Oswald, N., Krzywkowski, K.M., Friis, S., Jacobsen, R.B., Reuter, D., Taboryski, R., Kutchinsky, J., Vestergaard, R.K., Schroder, R.L., Sorensen, C.B., Bech, M., Korsgaard, M.P., and Willumsen, N.J. 2003. Upscaling and automation of electrophysiology: Toward high throughput screening in ion channel drug discovery. Receptors Channels 9:49‐58.
  Bell, J.E. and Miller, C. 1984. Effects of phospholipid surface charge on ion conduction in the K+ channel of sarcoplasmic reticulum. Biophys. J. 45:279‐287.
  Bliss, T.V. and Lomo, T. 1973. Long‐lasting potentiation of synaptic transmission in the dentate area of the anaesthetized rabbit following stimulation of the perforant path. J. Physiol. 232:331‐356.
  Bortolotto, Z.A., Amici, M., Anderson, W.W., Isaac, J.T., and Collingridge, G.L. 2011. Synaptic plasticity in the hippocampal slice preparation. Curr. Protoc. Neurosci. 54:6.13.1‐6.13.26.
  Bruggemann, A., George, M., Klau, M., Beckler, M., Steindl, J., Behrends, J.C., and Fertig, N. 2003. High quality ion channel analysis on a chip with the NPC technology. Assay Drug Devel. Technol. 1:665‐673.
  Campbell, I.G. 2009. EEG recording and analysis for sleep research. Curr. Protoc. Neurosci. 49:10.2.1‐10.2.19.
  Caspi, O., Itzhaki, I., Kehat, I., Gepstein, A., Arbel, G., Huber, I., Satin, J., and Gepstein, L. 2009. In vitro electrophysiological drug testing using human embryonic stem cell derived cardiomyocytes. Stem Cells Devel. 18:161‐172.
  Dascal, N. 2001. Voltage clamp recordings from Xenopus oocytes. Curr. Protoc. Neurosci. 10:6.12.1‐6.12.20.
  Daw, M. and Isaac, J. 2007. Electrophysiological recordings from neonatal neocortical brain slices. Curr. Protoc. Neurosci. 38: 6.23.1‐6.23.12.
  del Campo, C.M., Velazquez, J.L., and Freire, M.A. 2009. EEG recording in rodents, with a focus on epilepsy. Curr. Protoc. Neurosci. 49:6.24.1‐6.24.24.
  Dick, E., Rajamohan, D., Ronksley, J., and Denning, C. 2010. Evaluating the utility of cardiomyocytes from human pluripotent stem cells for drug screening. Biochem. Soc. Transact. 38:1037‐1045.
  Dunlop, J., Bowlby, M., Peri, R., Vasilyev, D., and Arias, R. 2008. High‐throughput electrophysiology: An emerging paradigm for ion‐channel screening and physiology. Nat. Rev. Drug Discov. 7:358‐368.
  El‐Arabi, A.M., Salazar, C.S., and Schmidt, J.J. 2012. Ion channel drug potency assay with an artificial bilayer chip. Lab Chip 12:2409‐2413.
  Farre, C., Stoelzle, S., Haarmann, C., George, M., Bruggemann, A., and Fertig, N. 2007. Automated ion channel screening: Patch clamping made easy. Exp. Opin. Therapeut. Targets 11:557‐565.
  Fejtl, M., Czubayko, U., Hümmer, A., Krauter, T., and Lepple‐Wienhues, A. 2007. Flip‐the‐tip: Automated patch clamping based on glass electrodes. Methods Mol. Biol. 403:71‐85.
  Finkel, A., Wittel, A., Yang, N., Handran, S., Hughes, J., and Costantin, J. 2006. Population patch clamp improves data consistency and success rates in the measurement of ionic currents. J. Biomol. Screen. 11:488‐496.
  Gillie, D.J., Novick, S.J., Donovan, B.T., Payne, L.A., and Townsend, C. 2013. Development of a high‐throughput electrophysiological assay for the human ether‐à‐go‐go related potassium channel hERG. J. Pharmacol. Toxicol. Methods 67:33‐44.
  Graef, J.D., Benson, L.C., Sidach, S.S., Wei, H., Lippiello, P.M., Bencherif, M., and Fedorov, N.B. 2013. Validation of a high‐throughput, automated electrophysiology platform for the screening of nicotinic agonists and antagonists. J. Biomol. Screen. 18:116‐127.
  Hamill, O.P., Marty, A., Neher, E., Sakmann, B., and Sigworth, F.J. 1981. Improved patch‐clamp techniques for high‐resolution current recording from cells and cell‐free membrane patches. Pflugers Arch. 391:85‐100.
  Hille, B. 2001. Ion Channels of Excitable Membranes. Sinauer Associates, Sunderland, Mass.
  Hodgkin, A.L. and Huxley, A.F. 1952a. The components of membrane conductance in the giant axon of Loligo. J. Physiol. 116:473‐496.
  Hodgkin, A.L. and Huxley, A.F. 1952b. Currents carried by sodium and potassium ions through the membrane of the giant axon of Loligo. J. Physiol. 116:449‐472.
  Hodgkin, A.L. and Huxley, A.F. 1952c. The dual effect of membrane potential on sodium conductance in the giant axon of Loligo. J. Physiol. 116:497‐506.
  Hodgkin, A.L., Huxley, A.F., and Katz, B. 1952. Measurement of current‐voltage relations in the membrane of the giant axon of Loligo. J. Physiol. 116:424‐448.
  Hogg, R.C. and Bertrand, D. 2004. Nicotinic acetylcholine receptors as drug targets. Curr. Drug Targets CNS Neurol. Disord. 3:123‐130.
  Horn, R. and Marty, A. 1988. Muscarinic activation of ionic currents measured by a new whole‐cell recording method. J. Gen. Physiol. 92:145‐159.
  Hubel, D.H. and Wiesel, T.N. 1965. Receptive fields and functional architecture in two nonstriate visual areas (18 and 19) of the cat. J. Neurophysiol. 28:229‐289.
  Illes, S., Fleischer, W., Siebler, M., Hartung, H.P., and Dihne, M. 2007. Development and pharmacological modulation of embryonic stem cell‐derived neuronal network activity. Exp. Neurol. 207:171‐176.
  International Human Genome Sequencing Consortium. 2004. Finishing the euchromatic sequence of the human genome. Nature 431:931‐945.
  Jackson, M.B. 2001a. Whole‐cell voltage clamp recording. Curr. Protoc. Neurosci. 00:6.6.1‐6.6.30.
  Jackson, M.B. 2001b. Single‐channel recording. Curr. Protoc. Neurosci. 2:6.8.1‐6.8.32.
  Klemic, K.G., Klemic, J.F., Reed, M.A., and Sigworth, F.J. 2002. Micromolded PDMS planar electrode allows patch clamp electrical recordings from cells. Biosens. Bioelectron. 17:597‐604.
  Kodandaramaiah, S.B., Franzesi, G.T., Chow, B.Y., Boyden, E.S., and Forest, C.R. 2012. Automated whole‐cell patch‐clamp electrophysiology of neurons in vivo. Nat. Methods 9:585‐587.
  Kostyuk, P.G., Krishtal, O.A., and Pidoplichko, V.I. 1975. Effect of internal fluoride and phosphate on membrane currents during intracellular dialysis of nerve cells. Nature 257:691‐693.
  Kreir, M., Farre, C., Beckler, M., George, M., and Fertig, N. 2008. Rapid screening of membrane protein activity: Electrophysiological analysis of OmpF reconstituted in proteoliposomes. Lab Chip 8:587‐595.
  Lander, E.S., Linton, L.M., Birren, B., Nusbaum, C., Zody, M.C., Baldwin, J., Devon, K., Dewar, K., Doyle, M., FitzHugh, W., Funke, R., Gage, D., Harris, K., Heaford, A., Howland, J., Kann, L., Lehoczky, J., LeVine, R., McEwan, P., McKernan, K., Meldrim, J., Mesirov, J.P., Miranda, C., Morris, W., Naylor, J., Raymond, C., Rosetti, M., Santos, R., Sheridan, A., Sougnez, C., Stange‐Thomann, N., Stojanovic, N., Subramanian, A., Wyman, D., Rogers, J., Sulston, J., Ainscough, R., Beck, S., Bentley, D., Burton, J., Clee, C., Carter, N., Coulson, A., Deadman, R., Deloukas, P., Dunham, A., Dunham, I., Durbin, R., French, L., Grafham, D., Gregory, S., Hubbard, T., Humphray, S., Hunt, A., Jones, M., Lloyd, C., McMurray, A., Matthews, L., Mercer, S., Milne, S., Mullikin, J.C., Mungall, A., Plumb, R., Ross, M., Shownkeen, R., Sims, S., Waterston, R.H., Wilson, R.K., Hillier, L.W., McPherson, J.D., Marra, M.A., Mardis, E.R., Fulton, L.A., Chinwalla, A.T., Pepin, K.H., Gish, W.R., Chissoe, S.L., Wendl, M.C., Delehaunty, K.D., Miner, T.L., Delehaunty, A., Kramer, J.B., Cook, L.L., Fulton, R.S., Johnson, D.L., Minx, P.J., Clifton, S.W., Hawkins, T., Branscomb, E., Predki, P., Richardson, P., Wenning, S., Slezak, T., Doggett, N., Cheng, J.F., Olsen, A., Lucas, S., Elkin, C., Uberbacher, E., Frazier, M., Gibbs, R.A., Muzny, D.M., Scherer, S.E., Bouck, J.B., Sodergren, E.J., Worley, K.C., Rives, C.M., Gorrell, J.H., Metzker, M.L., Naylor, S.L., Kucherlapati, R.S., Nelson, D.L., Weinstock, G.M., Sakaki, Y., Fujiyama, A., Hattori, M., Yada, T., Toyoda, A., Itoh, T., Kawagoe, C., Watanabe, H., Totoki, Y., Taylor, T., Weissenbach, J., Heilig, R., Saurin, W., Artiguenave, F., Brottier, P., Bruls, T., Pelletier, E., Robert, C., Wincker, P., Smith, D.R., Doucette‐Stamm, L., Rubenfield, M., Weinstock, K., Lee, H.M., Dubois, J., Rosenthal, A., Platzer, M., Nyakatura, G., Taudien, S., Rump, A., Yang, H., Yu, J., Wang, J., Huang, G., Gu, J., Hood, L., Rowen, L., Madan, A., Qin, S., Davis, R.W., Federspiel, N.A., Abola, A.P., Proctor, M.J., Myers, R.M., Schmutz, J., Dickson, M., Grimwood, J., Cox, D.R., Olson, M.V., Kaul, R., Raymond, C., Shimizu, N., Kawasaki, K., Minoshima, S., Evans, G.A., Athanasiou, M., Schultz, R., Roe, B.A., Chen, F., Pan, H., Ramser, J., Lehrach, H., Reinhardt, R., McCombie, W.R., de la Bastide, M., Dedhia, N., Blocker, H., Hornischer, K., Nordsiek, G., Agarwala, R., Aravind, L., Bailey, J.A., Bateman, A., Batzoglou, S., Birney, E., Bork, P., Brown, D.G., Burge, C.B., Cerutti, L., Chen, H.‐C., Church, D., Clamp, M., Copley, R.R., Doerks, T., Eddy, S.R., Eichler, E.E., Furey, T.S., Galagan, J., Gilbert, J.G., Harmon, C., Hayashizaki, Y., Haussler, D., Hermjakob, H., Hokamp, K., Jang, W., Johnson, L.S., Jones, T.A., Kasif, S., Kaspryzk, A., Kennedy, S., Kent, W.J., Kitts, P., Koonin, E.V., Korf, I., Kulp, D., Lancet, D., Lowe, T.M., McLysaght, A., Mikkelsen, T., Moran, J.V., Mulder, N., Pollara, V.J., Ponting, C.P., Schuler, G., Schultz, J., Slater, G., Smit, A.F., Stupka, E., Szustakowski, J., Thierry‐Mieg, D., Thierry‐Mieg, J., Wagner, L., Wallis, J., Wheeler, R., Williams, A., Wolf, Y.I., Wolfe, K.H., Yang, S.‐P., Yeh, R.‐F., Collins, F., Guyer, M.S., Peterson, J., Felsenfeld, A., Wetterstrand, K.A., Patrinos, A., Morgan, M.J., de Jong, P., Catanese, J.J., Osoegawa, K., Shizuya, H., Choi, S., Chen, Y.J., and International Human Genome Sequencing Consortium. 2001. Initial sequencing and analysis of the human genome. Nature 409:860‐921.
  Latorre, R., Vergara, C., and Hidalgo, C. 1982. Reconstitution in planar lipid bilayers of a Ca2+‐dependent K+ channel from transverse tubule membranes isolated from rabbit skeletal muscle. Proc. Natl. Acad. Sci. U.S.A. 79:805‐809.
  Light, G.A., Williams, L.E., Minow, F., Sprock, J., Rissling, A., Sharp, R., Swerdlow, N.R., and Braff, D.L. 2010. Electroencephalography (EEG) and event‐related potentials (ERPs) with human participants. Curr. Protoc. Neurosci. 52:6.25.1‐6.25.24.
  Liu, Q., Huang, H., Cai, H., Xu, Y., Li, Y., Li, R., and Wang, P. 2007. Embryonic stem cells as a novel cell source of cell‐based biosensors. Biosens. Bioelectron. 22:810‐815.
  Madison, D.V. and Edson, E.B. 2001. Preparation of hippocampal brain slices. Curr. Protoc. Neurosci. 00:6.4.1‐6.4.7.
  Mathes, C. 2006. QPatch: The past, present and future of automated patch clamp. Exp. Opin. Therapeut. Targets 10:319‐327.
  McConnell, E.R., McClain, M.A., Ross, J., Lefew, W.R., and Shafer, T.J. 2012. Evaluation of multi‐well microelectrode arrays for neurotoxicity screening using a chemical training set. Neurotoxicology 33:1048‐1057.
  Mueller, P., Rudin, D.O., Tien, H.T., and Wescott, W.C. 1962. Reconstitution of cell membrane structure in vitro and its transformation into an excitable system. Nature 194:979‐980.
  Narahashi, T. 2003. Principles of electrophysiology: An overview. Curr. Protoc. Toxicol. 17:11.10.1‐11.10.11.
  Neher, E. and Sakmann, B. 1976. Single‐channel currents recorded from membrane of denervated frog muscle fibres. Nature 260:799‐802.
  Noda, M., Takahashi, H., Tanabe, T., Toyosato, M., Furutani, Y., Hirose, T., Asai, M., Inayama, S., Miyata, T., and Numa, S. 1982. Primary structure of alpha‐subunit precursor of Torpedo californica acetylcholine receptor deduced from cDNA sequence. Nature 299:793‐797.
  Novak, J.L. and Wheeler, B.C. 1988. Multisite hippocampal slice recording and stimulation using a 32 element microelectrode array. J. Neurosci. Methods 23:149‐159.
  Oka, H., Shimono, K., Ogawa, R., Sugihara, H., and Taketani, M. 1999. A new planar multielectrode array for extracellular recording: Application to hippocampal acute slice. J. Neurosci. Methods 93:61‐67.
  Papke, R.L. and Smith‐Maxwell, C. 2009. High throughput electrophysiology with Xenopus oocytes. Comb. Chem. High Throughput Screen. 12:38‐50.
  Peng, S., Lacerda, A.E., Kirsch, G.E., Brown, A.M., and Bruening‐Wright, A. 2010. The action potential and comparative pharmacology of stem cell‐derived human cardiomyocytes. J. Pharmacol. Toxicol. Methods 61:277‐286.
  Pihl, J., Sinclair, J., Karlsson, M., and Orwar, O. 2005. Microfluidics for cell‐based assays. Materials Today 8:46‐51.
  Sartipy, P., Björquist, P., Strehl, R., and Hyllner, J. 2007. The application of human embryonic stem cell technologies to drug discovery. Drug Discov. Today 12:688‐699.
  Schäfers, M. and Cain, D. 2004. Single‐fiber recording: In vivo and in vitro preparations. Methods Mol. Med. 99:155‐166.
  Schmelz, M. and Schmidt, R. 2010. Microneurographic single‐unit recordings to assess receptive properties of afferent human C‐fibers. Neurosci. Lett. 470:158‐161.
  Schnizler, K., Kuster, M., Methfessel, C., and Fejtl, M. 2003. The roboocyte: Automated cDNA/mRNA injection and subsequent TEVC recording on Xenopus oocytes in 96‐well microtiter plates. Receptors Channels 9:41‐48.
  Schroeder, K., Neagle, B., Trezise, D.J., and Worley, J. 2003. Ionworks HT: A new high‐throughput electrophysiology measurement platform. J. Biomol. Screen. 8:50‐64.
  Serra, J., Bostock, H., and Navarro, X. 2010. Microneurography in rats: A minimally invasive method to record single C‐fiber action potentials from peripheral nerves in vivo. Neurosci. Lett. 470:168‐174.
  Shieh, C.C., Trumbull, J.D., Sarthy, J.F., McKenna, D.G., Parihar, A.S., Zhang, X.F., Faltynek, C.R., and Gopalakrishnan, M. 2003. Automated Parallel Oocyte Electrophysiology Test station (POETs): A screening platform for identification of ligand‐gated ion channel modulators. Assay Drug Devel. Technol. 1:655‐663.
  Spencer, C.I., Li, N., Chen, Q., Johnson, J., Nevill, T., Kammonen, J., and Ionescu‐Zanetti, C. 2012. Ion channel pharmacology under flow: Automation via well‐plate microfluidics. Assay Drug Devel. Technol. 10:313‐324.
  Stastna, M., Abraham, M.R., and Van Eyk, J.E. 2009. Cardiac stem/progenitor cells, secreted proteins, and proteomics. FEBS Lett. 583:1800‐1807.
  Stuart, G. 2001. Patch‐pipet recording in brain slices. Curr. Protoc. Neurosci. 2:6.7.1‐6.7.10.
  Swanson, G.T. and Contractor, A. 2004. Recording in the cerebellar slice. Curr. Protoc. Neurosci. 25:6.18.1‐6.18.14.
  Szymusiak, R. and Nitz, D. 2003. Chronic recording of extracellular neuronal activity in behaving animals. Curr. Protoc. Neurosci. 2:6.16.1‐6.16.24
  Tao, H., Santa Ana, D., Guia, A., Huang, M., Ligutti, J., Walker, G., Sithiphong, K., Chan, F., Guoliang, T., Zozulya, Z., Saya, S., Phimmachack, R., Sie, C., Yuan, J., Wu, L., Xu, J., and Ghetti, A. 2004. Automated tight seal electrophysiology for assessing the potential hERG liability of pharmaceutical compounds. Assay Drug Devel. Technol. 2:497‐506.
  Terstappen, G.C., Roncarati, R., Dunlop, J., and Peri, R. 2010. Screening technologies for ion channel drug discovery. Future Med. Chem. 2:715‐730.
  Ussing, H.H. and Zerahn, K. 1951. Active transport of sodium as the source of electric current in the short‐circuited isolated frog skin. Acta Physiol. Scand. 23:110‐127.
  Venter, J.C., Adams, M.D., Myers, E.W., Li, P.W., Mural, R.J., Sutton, G.G., Smith, H.O., Yandell, M., Evans, C.A., Holt, R.A., Gocayne, J.D., Amanatides, P., Ballew, R.M., Huson, D.H., Wortman, J.R., Zhang, Q., Kodira, C.D., Zheng, X.H., Chen, L., Skupski, M., Subramanian, G., Thomas, P.D., Zhang, J., Gabor Miklos, G.L., Nelson, C., Broder, S., Clark, A.G., Nadeau, J., McKusick, V.A., Zinder, N., Levine, A.J., Roberts, R.J., Simon, M., Slayman, C., Hunkapiller, M., Bolanos, R., Delcher, A., Dew, I., Fasulo, D., Flanigan, M., Florea, L., Halpern, A., Hannenhalli, S., Kravitz, S., Levy, S., Mobarry, C., Reinert, K., Remington, K., Abu‐Threideh, J., Beasley, E., Biddick, K., Bonazzi, V., Brandon, R., Cargill, M., Chandramouliswaran, I., Charlab, R., Chaturvedi, K., Deng, Z., Di Francesco, V., Dunn, P., Eilbeck, K., Evangelista, C., Gabrielian, A.E., Gan, W., Ge, W., Gong, F., Gu, Z., Guan, P., Heiman, T.J., Higgins, M.E., Ji, R.R., Ke, Z., Ketchum, K.A., Lai, Z., Lei, Y., Li, Z., Li, J., Liang, Y., Lin, X., Lu, F., Merkulov, G.V., Milshina, N., Moore, H.M., Naik, A.K., Narayan, V.A., Neelam, B., Nusskern, D., Rusch, D.B., Salzberg, S., Shao, W., Shue, B., Sun, J., Wang, Z., Wang, A., Wang, X., Wang, J., Wei, M.‐H., Wides, R., Xiao, C., Yan, C., Yao, A., Ye, J., Zhan, M., Zhang, W., Zhang, H., Zhao, Q., Zheng, L., Zhong, F., Zhong, W., Zhu, S., Zhao, S., Gilbert, D., Baumhueter, S., Spier, G., Carter, C., Cravchik, A., Woodage, T., Ali, F., An, H., Awe, A., Baldwin, D., Baden, H., Barnstead, M., Barrow, I., Beeson, K., Busam, D., Carver, A., Center, A., Cheng, M.L., Curry, L., Danaher, S., Davenport, L., Desilets, R., Dietz, S., Dodson, K., Doup, L., Ferriera, S., Garg, N., Gluecksmann, A., Hart, B., Haynes, J., Haynes, C., Heiner, C., Hladun, S., Hostin, D., Houck, J., Howland, T., Ibegwam, C., Johnson, J., Kalush, F., Kline, L., Koduru, S., Love, A., Mann, F., May, D., McCawley, S., McIntosh, T., McMullen, I., Moy, M., Moy, L., Murphy, B., Nelson, K., Pfannkoch, C., Pratts, E., Puri, V., Qureshi, H., Reardon, M., Rodriguez, R., Rogers, Y.‐H., Romblad, D., Ruhfel, B., Scott, R., Sitter, C., Smallwood, M., Stewart, E., Strong, R., Suh, E., Thomas, R., Tint, N.N., Tse, S., Vech, C., Wang, G., Wetter, J., Williams, S., Williams, M., Windsor, S., Winn‐Deen, E., Wolfe, K., Zaveri, J., Zaveri, K., Abril, J.F., Guigo, R., Campbell, M.J., Sjolander, K.V., Karlak, B., Kejariwal, A., Mi, H., Lazareva, B., Hatton, T., Narechania, A., Diemer, K., Muruganujan, A., Guo, N., Sato, S., Bafna, V., Istrail, S., Lippert, R., Schwartz, R., Walenz, B., Yooseph, S., Allen, D., Basu, A., Baxendale, J., Blick, L., Caminha, M., Carnes‐Stine, J., Caulk, P., Chiang, Y.‐H., Coyne, M., Dahlke, C., Mays, A., Dombroski, M., Donnelly, M., Ely, D., Esparham, S., Fosler, C., Gire, H., Glanowski, S., Glasser, K., Glodek, A., Gorokhov, M., Graham, K., Gropman, B., Harris, M., Heil, J., Henderson, S., Hoover, J., Jennings, D., Jordan, C., Jordan, J., Kasha, J., Kagan, L., Kraft, C., Levitsky, A., Lewis, M., Liu, X., Lopez, J., Ma, D., Majoros, W., McDaniel, J., Murphy, S., Newman, M., Nguyen, T., Nguyen, N., Nodell, M., Pan, S., Peck, J., Peterson, M., Rowe, W., Sanders, R., Scott, J., Simpson, M., Smith, T., Sprague, A., Stockwell, T., Turner, R., Venter, E., Wang, M., Wen, M., Wu, D., Wu, M., Xia, A., Zandieh, A., and Zhu, X. 2001. The sequence of the human genome. Science 291:1304‐1351.
  Witten, I.B., Lin, S.C., Brodsky, M., Prakash, R., Diester, I., Anikeeva, P., Gradinaru, V., Ramakrishnan, C., and Deisseroth, K. 2010. Cholinergic interneurons control local circuit activity and cocaine conditioning. Science 330:1677‐1681.
  Yazawa, M., Hsueh, B., Jia, X., Pasca, A.M., Bernstein, J.A., Hallmayer, J., and Dolmetsch, R.E. 2011. Using induced pluripotent stem cells to investigate cardiac phenotypes in Timothy syndrome. Nature 471:230‐234.
  Zhang, F., Gradinaru, V., Adamantidis, A.R., Durand, R., Airan, R.D., de Lecea, L., and Deisseroth, K. 2010. Optogenetic interrogation of neural circuits: Technology for probing mammalian brain structures. Nat. Protoc. 5:439‐456.
  Zou, B., Yu, H., Babcock, J.J., Chanda, P., Bader, J.S., McManus, O.B., and Li, M. 2010. Profiling diverse compounds by flux‐ and electrophysiology‐based primary screens for inhibition of human Ether‐a‐go‐go related gene potassium channels. Assay Drug Devel. Technol. 8:743‐754.
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