eBriefing 
Kavli Workshop in Pucon
Presented By
Overview
The technology to eavesdrop on exocytosis events is improving and bringing new insights to our understanding of cellular communication. Thanks to a generous grant from the Kavli Institute at Cornell for Nanoscale Science and with support from HEKA Electronic, a workshop entitled Advanced Techniques to Study Single Vesicle Exocytosis and a more informal W&BC (wine and beer club) on Advanced Instrumentation for these studies, were held during the 13th International Symposium on Chromaffin Cell Biology in Pucón, Chile, which took place January 7–12, 2006. Ana Cárdenas of Valparaíso University organized an outstanding meeting in a spectacular environment between Lake Villarica and the impressive Villarrica Volcano. The meeting attracted 120 attendees from about 15 countries who had very productive discussions on many topics related to exocytosis, ranging from molecular to clinical aspects.
Web Sites
Sulzer Lab Software and Models
Sulzer lab software and models, including spreadsheets for random walks.
HEKA Elektronik
Includes HEKA PATCHMASTER specifications.
Gillis Electrophysiology Lab
The homesite of Kevin Gillis at the University of Missouri has information about current projects, including images and descriptions.
Lindau Research Group
Manfred Lindau's team at Cornell University.
Journal Articles
Analyzing the Single Vesicle Spike
Alés, E., L. Tabares, J. M. Poyato, et al. 1999. High calcium concentrations shift the mode of exocytosis to the kiss-and-run mechanism. Nat. Cell Biol. 1: 40-44.
Amatore, C., Y. Bouret, E. R. Travis & R. M. Wightman. 2000. Interplay between membrane dynamics, diffusion and swelling pressure governs individual vesicular exocytotic events during release of adrenaline by chromaffin cells. Biochimie 82: 481-496.
Berg, H. C. 1983. Random Walks in Biology. Princeton University Press, Princeton, NJ.
Chow, R. H. & L. von Ruden. 1995. Electrochemical detection of secretion from single cells. In Single-Channel Recording. B. Sakmann & E. Neher, Eds. Plenum Press, New York.
Chow, R. H., L. von Ruden & E. Neher. 1992. Delay in vesicle fusion revealed by electrochemical monitoring of single secretory events in adrenal chromaffin cells. Nature 356: 60-63.
Crank, J. 1975. The Mathematics of Diffusion. Oxford University Press, Oxford, UK.
Elhamdani, A., F. Azizi & C. R. Artalejo. 2006. Double patch clamp reveals that transient fusion (kiss-and-run) is a major mechanism of secretion in calf adrenal chromaffin cells: high calcium shifts the mechanism from kiss-and-run to complete fusion. J. Neurosci. 26: 3030-3036.
Gonon, F. R. Cespuglio, J. L. Ponchon, et al. 1978. In vivo continuous electrochemical determination of dopamine release in rat neostriatum. C. R. Acad. Sci. Hebd. Seances Acad. Sci. D 286: 1203-1206.
Marszalek, P. B. Farell & J. M. Fernandez. 1996. Ion-exchange gel regulates neurotransmitter release through the exocytotic fusion pore. Soc. Gen. Physiol. Ser 51: 211-222.
Mosharov, E. V. & D. Sulzer. 2005. Analysis of exocytotic events recorded by amperometry. Nat. Methods 2: 651-658.
Schroeder, T. J., J. A. Jankowski, K. T. Kawagoe, et al. 1992. Analysis of diffusional broadening of vesicular packets of catecholamines released from biological cells during exocytosis. Anal. Chem. 64: 3077-3083.
Staal, R. G., E. V. Mosharov & D. Sulzer. 2004. Dopamine neurons release transmitter via a flickering fusion pore. Nat. Neurosci. 7: 341-346.
Caged Calcium and Its Measurement: Exocytosis in a Flash
Bollmann, J. H., B. Sakmann & J. G. Borst. 2000. Calcium sensitivity of glutamate release in a calyx-type terminal. Science 289: 953-957.
Ellis-Davies, G. C. & R. J. Barsotti. 2006. Tuning caged calcium: photolabile analogues of EGTA with improved optical and chelation properties. Cell Calcium 39: 75-83.
Ellis-Davies, G. C. & J. H. Kaplan. 1994. Nitrophenyl-EGTA, a photolabile chelator that selectively binds Ca2+ with high affinity and releases it rapidly upon photolysis. Proc. Natl. Acad. Sci. USA 91: 187-191. Full Text
Heinemann, C., R. H. Chow, E. Neher & R. S. Zucker. 1994. Kinetics of the secretory response in bovine chromaffin cells following flash photolysis of caged Ca2+. Biophys. J. 67: 2546-2557. Full Text
Kaplan, J. H. & G. C. Ellis-Davies. 1988. Photolabile chelators for the rapid photorelease of divalent cations. Proc. Natl. Acad. Sci. USA 85: 6571-6575. Full Text
Momotake, A., N. Lindegger, E. Niggli, et al. 2006. The nitrodibenzofuran chromophore: a new caging group for ultra-efficient photolysis in living cells. Nat. Methods 3: 35-40.
Naraghi, M., T. H. Muller & E. Neher. 1998. Two-dimensional determination of the cellular Ca2+ binding in bovine chromaffin cells. Biophys. J. 75: 1635-1647. Full Text
Schneggenburger, R. & E. Neher. 2000. Intracellular calcium dependence of transmitter release rates at a fast central synapse. Nature 406: 8.
Soeller, C., M. D. Jacobs, K. T. Jones, et al. 2003. Application of two-photon flash photolysis to reveal intercellular communication and intracellular Ca2+ movements. J. Biomed. Opt. 8: 418-427.
Sørensen, J. B., U. Matti, S. H. Wei et al. 2002. The SNARE protein SNAP-25 is linked to fast calcium triggering of exocytosis. Proc. Natl. Acad. Sci. USA 99: 1627-1632. Full Text
The Nanomechanics of Endocytosis and Exocytosis in Live Cells
Jahn, R., T. Lang & T. C. Sudhof. 2003. Membrane fusion. Cell 112: 519-533.
Lang, T., D. Bruns, D. Wenzel, et al. 2001. SNAREs are concentrated in cholesterol-dependent clusters that define docking and fusion sites for exocytosis. Embo. J. 20: 2202-2213. Full Text
Merrifield, C. J., D. Perrais & D. Zenisek. 2005. Coupling between clathrin-coated-pit invagination, cortactin recruitment, and membrane scission observed in live cells. Cell 121: 593-606.
Merrifield, C. J., B. Qualmann, M. M. Kessels & W. Almers. 2004. Neural Wiskott Aldrich Syndrome Protein (N-WASP) and the Arp2/3 complex are recruited to sites of clathrin-mediated endocytosis in cultured fibroblasts. Eur. J. Cell Biol. 83: 13-18.
Steyer, J. A., H. Horstmann & W. Almers. 1997. Transport, docking and exocytosis of single secretory granules in live chromaffin cells. Nature 388: 474-478.
Zenisek, D., J. A. Steyer & W. Almers. 2000. Transport, capture and exocytosis of single synaptic vesicles at active zones. Nature 406: 849-854.
A Software Lock-in Amplifier for On-Cell Membrane Capacitance Measurements
Debus, K. & M. Lindau. 2000. Resolution of patch capacitance recordings and of fusion pore conductances in small vesicles. Biophys. J. 78: 2983-2997. Full Text
Gillis, K. D. 2000. Admittance-based measurement of membrane capacitance using the EPC-9 patch-clamp amplifier. Pflügers Arch. 439: 655-664.
Gillis, K. D. 1995. Techniques for membrane capacitance measurements. In Single-Channel Recording, 2nd Edition. B. Sakmann & E. Neher, Eds. Plenum, New York.
Lindau, M. & E. Neher. 1988. Patch-clamp techniques for time-resolved capacitance measurements in single cells. Pflügers Arch. 411: 137-146
Neher, E. & A. Marty. 1982. Discrete changes of cell membrane capacitance observed under conditions of enhanced secretion in bovine adrenal chromaffin cells. Proc. Natl. Acad. Sci. USA 79: 6712-6716. Full Text
Exocytosis on the Chip
Albillos, A., G. Dernick, H. Horstman, et al. 1997. The exocytotic event in chromaffin cells revealed by patch amperometry. Nature 389: 509-512.
An, S. J. & W. Almers. 2004. Tracking SNARE complex formation in live endocrine cells. Science 306: 1042-1046.
Breckenridge, L. J. & W. Almers. 1987. Currents through the fusion pore that forms during exocytosis of a secretory vesicle. Nature 328: 814-817.
Chow, R. H., L. Von Ruden & E. Neher. 1992. Delay in vesicle fusion revealed by electrochemical monitoring of single secretory events in adrenal chromaffin cells. Nature 356: 60-63.
Debus, K. & M. Lindau. 2000. Resolution of patch capacitance recordings and of fusion pore conductances in small vesicles. Biophys. J. 78: 2983-2997. Full Text
Dernick, G., G. Alvarez De Toledo & M. Londau. 2003. Exocytosis of single chromaffin granules in cell-free inside-out membrane patches. Nat. Cell Biol. 5: 358-362.
Dernick, G., L. W. Gong, L. Tabares, et al. 2005. Patch amperometry: high-resolution measurements of single-vesicle fusion and release. Nat. Methods 2: 699-708.
Dias, A. F., G. Dernick, V. Valero, et al. 2002. Growth of carbon nanotubes by thermal and plasma chemical vapour deposition processes and applications in microscopy. Nanotechnology 13: 285-289.
Hafez, I., K. Kisler, K. Berberian, et al. 2005. Electrochemical imaging of fusion pore openings by electrochemical detector arrays. Proc. Natl. Acad. Sci. USA 102: 13879-13884. Full Text
Sigworth, F. J. & K. G. Klemic. 2005. Microchip technology in ion-channel research. IEEE Trans. Nanobioscience 4: 121-127.
Söllner, T., S. Whiteheart, M. Brunner, et al. 1993. SNAP receptors implicated in vesicle targeting and fusion. Nature 362, 318-323.
Wightman, R. M., J. A. Jankowski, R. T. Kennedy, et al. 1991. Temporally resolved catecholamine spikes correspond to single vesicle release from individual chromaffin cells. Proc. Natl. Acad. Sci. USA 88: 10754-10758. (PDF, 1.05 MB) Full Text
Studies of Exocytosis from Chromaffin Cells Using Microchip Electrochemistry and Scanning Ion Conductance Microscopy
Burgoyne, R. D. & A. Morgan. 2003. Secretory ganule eocytosis. Physiol. Rev. 83: 581-632. Full Text
Chow, R. H., L. Von Ruden & E. Neher. 1992. Delay in vesicle fusion revealed by electrochemical monitoring of single secretory events in adrenal chromaffin cells. Nature 356: 60-63.
Duffy, D. C., J. C. McDonald, O. J. A. Schueller & G. M. Whitesides. 1998. Rapid prototyping of microfluidic systems in poly(dimethylsiloxane). Anal. Chem. 70: 4974-4984.
Hansma, P. K., B. Drake, O. Marti, et al. 1989. The scanning ion-conductance microscope. Science 243: 641-643.
Hayashi, K., Y. Iwasaki, T. Horiuchi, et al. 2005. Selective detection of a catecholamine against electroactive interferents using an interdigitated heteroarray electrode consisting of a metal oxide electrode and a metal band electrode. Anal. Chem. 77: 5236-5242.
Korchev, Y. E., Y. A. Negulyaev, C. R. Edwards, et al. 2000. Functional localization of single active ion channels on the surface of a living cell. Nat. Cell Biol. 2: 616-619.
Qiu, H., J. Yan, X. Sun, et al. 2003. Microchip capillary electrophoresis with an integrated indium tin oxide electrode-based electrochemiluminescence detector. Anal. Chem. 75: 5435-5440.
Schroeder, T. J., J. A. Jankowski, K. T. Kawagoe, et al. 1992. Analysis of diffusional broadening of vesicular packets of catecholamines released from biological cells during exocytosis. Anal. Chem. 64: 3077-3083.
Sun, X. & K. D. Gillis. 2006. On-chip amperometric measurement of quantal catecholamine release using transparent indium tin oxide electrodes. Anal. Chem. 78: 2521-2525.
Wightman, R. M., J. A. Jankowski, R. T. Kennedy, et al. 1991. Temporally resolved catecholamine spikes correspond to single vesicle release from individual chromaffin cells. Proc. Natl. Acad. Sci. USA 88: 10754-10758. (PDF, 1.05 MB) Full Text
Zhan, W., J. Alvarez & R. M. Crooks. 2002. Electrochemical sensing in microfluidic systems using electrogenerated chemiluminescence as a photonic reporter of redox reactions. J. Am. Chem. Soc. 124: 13265-13270.
Organizers
Ricardo Borges, MD, PhD
La Laguna University, Spain
email | publications
Ricardo Borges is Professor Titular of Pharmacology at the Medical School of La Laguna (Tenerife, Spain) where he leads a laboratory whose work focuses on neurosecretion. He has been applying amperometrical techniques to explore the secretory responses of adrenergic tissues since 1985. His research interest is the modulation of the latest steps of exocytosis.
Borges received his MD from the University of La Laguna in 1982 and his PhD from the University of Alicante in 1985. He was postdoc at the University of Massachusetts and King's College London. He was also a visiting professor in the chemistry department at the University of North Carolina.
Manfred Lindau
Cornell University
email | web site | publications
Manfred Lindau was trained as a physicist and received his doctorate from the Technical University of Berlin in 1983 in the field of physical chemistry. Lindau was a postdoctoral associate at the Max-Planck-Institute for Biophysical Chemistry in Göttingen and at the Free University of Berlin, where he became an assistant professor in 1988. From 1992 through 1997 he was an associate member of the Max-Planck-Institute for Medical Research and taught biophysics at the University of Heidelberg. He joined the faculty at Cornell University in 1997. He is a founding member, member of the executive committee, and program coordinator of the Nanoscale Cell Biology Program at the Science and Technology Center for Nanobiotechnology at Cornell. He is an elected member of the Asian Institute of NanoBioScience and Technology.
Lindau is one of the leading scientists in the field of exocytosis and endocytosis. He has developed and applied biophysical techniques that allow investigation of single vesicle exocytosis and endocytosis in unprecedented detail. In 2003 he received a Research Award from the Alexander von Humboldt Foundation, Germany in recognition of his scientific achievements. The citation index lists more than 1500 citations of his publications. Lindau has given over 90 invited lectures at conferences and seminars around the world. He is active as a consultant in the areas of biophysics, physiology, and cell biology, and is a member of the Biophysical Society and of the Society for Neuroscience.
Speakers
David Sulzer, PhD
Columbia University
email | web site | publications
Eugene Mosharov, PhD
Columbia University
email | web site | publications
Gustavo Rivas
National University of Cordoba, Argentina
email | publications
Reinhard Jahn, PhD
Max-Planck-Instute for Biophysical Chemistry
email | web site | publications
Jakob Sørensen, PhD
Max-Planck-Institute for Biophysical Chemistry
email | web site | publications
Wolfhard Almers, PhD
Vollum Institute
Oregon Health & Science University
email | web site | publications
Christian Heinemann, PhD
HEKA Elektronik
email | web site
Kevin D. Gillis, PhD
University of Missouri
email | web site | publications