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Analysis of Single-Vesicle Exocytotic Events Recorded by Amperometry

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Exocytosis and Endocytosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 440))

Summary

Analysis of the shape of the amperometric spikes that originate from the oxidation of monoamine molecules released during the fusion of individual secretory vesicles provides valuable data about molecular steps involved in stimulation-dependent transmitter release. The sensitivity of amperometry permits the detection of the monoamines released from individual vesicles, giving information about intracellular transmitter homeostasis (synthesis, degradation, reuptake, etc.) and about the release probability (vesicle availability and sensitivity of fusion machinery to Ca+2). Furthermore, temporal resolution of amperometry allows for the observation of the real-time kinetics of monoamine release during exocytosis. Here, we provide algorithms for computerized analysis of the amperometric signal with emphasis on routines that confer consistent determination of spike parameters on recordings with different sampling rates, signal-to-noise levels, spike durations, and surrounding signal.

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References

  1. 1. Wightman, R.M., Jankowski, J.A., Kennedy, R.T., et al. (1991) Temporally resolved catecholamine spikes correspond to single vesicle release from individual chromaffin cells. Proc. Natl. Acad. Sci. U. S. A. 88, 10754–10758.

    Article  CAS  PubMed  Google Scholar 

  2. 2. Mosharov, E.V., and Sulzer, D. (2005) Analysis of exocytotic events recorded by amperometry. Nat. Methods 2, 651–658.

    Article  CAS  PubMed  Google Scholar 

  3. 3. Segura, F., Brioso, M.A., Gómez, J.F., Machado, J.D., and Borges, R. (2000) Automatic analysis for amperometrical recordings of exocytosis. J. Neurosci. Methods 103, 151–156.

    Article  CAS  PubMed  Google Scholar 

  4. 4. Kissinger, P.T., Hart, J.B., and Adams, R.N. (1973) Voltammetry in brain tissue—a new neurophysiological measurement. Brain Res. 55, 209–213.

    Article  CAS  PubMed  Google Scholar 

  5. 5. Chow, R.H., and von Ruden, L. (1995) In Single-Channel Recording (Sakmann, B., and Neher, E., eds.), Plenum Press, New York, pp. 245–276.

    Google Scholar 

  6. 6. Colliver, T., Hess, E.J., Pothos, E.N., Sulzer, D., and Ewing, A.G. (2000) Quantitative and statistical analysis of the shape of amperometric spikes recorded from two populations of cells. J. Neurochem. 74, 1086–1097.

    Article  CAS  PubMed  Google Scholar 

  7. 7. Wang, C.T., Grishanin, R., Earles, C.A., Chang, P.Y., Martin, T.F., Chapman, E.R., and Jackson, M.B. (2001) Synaptotagmin modulation of fusion pore kinetics in regulated exocytosis of dense-core vesicles. Science 294, 1111–1115.

    Article  CAS  PubMed  Google Scholar 

  8. 8. Sulzer, D., and Pothos, E.N. (2000) Regulation of quantal size by presynaptic mechanisms. Rev. Neurosci. 11, 159–212.

    CAS  PubMed  Google Scholar 

  9. 9. Ott, R.L., and Longnecker, M. (2001) An Introduction to Statistical Methods and Data Analysis, Duxbury Press, Belmont, CA.

    Google Scholar 

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Acknowledgments

I thank Drs. David Sulzer and Ricardo Borges for critique of the manuscript. This work was supported by grants from the Parkinson's Disease Foundation and Picower Foundation awarded to D. Sulzer.

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© 2008 Humana Press, a part of Springer Science+Business Media, LLC

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Mosharov, E.V. (2008). Analysis of Single-Vesicle Exocytotic Events Recorded by Amperometry. In: Ivanov, A.I. (eds) Exocytosis and Endocytosis. Methods in Molecular Biology, vol 440. Humana Press. https://doi.org/10.1007/978-1-59745-178-9_24

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  • DOI: https://doi.org/10.1007/978-1-59745-178-9_24

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-865-2

  • Online ISBN: 978-1-59745-178-9

  • eBook Packages: Springer Protocols

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