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The exocytotic event in chromaffin cells revealed by patch amperometry

Nature volume 389, pages 509–512 (1997)Cite this article

Abstract

In mast cells and granulocytes, exocytosis starts with the formation of a fusion pore1,2,3. It has been suggested that neurotransmitters may be released through such a narrow pore without full fusion4,5. However, owing to the small size of the secretory vesicles containing neurotransmitter, the properties of the fusion pore formed during Ca2+-dependent exocytosis and its role in transmitter release are still unknown. Here we investigate exocytosis of individual chromaffin granules by using cell-attached capacitance measurements3,6 combined with electrochemical detection of catecholamines7,8, achieved by inserting a carbon-fibre electrode into the patch pipette. This allows the simultaneous determination of the opening of individual fusion pores and of the kinetics of catecholamine release from the same vesicle. We found that the fusion-pore diameter stays at <3 nm for a variable period, which can last for several seconds, before it expands. Transmitter is released much faster through this pore than in mast cells, generating a ‘foot’ signal8 which precedes the amperometric spike. Occasionally, the narrow pore forms only transiently and does not expand, allowing complete transmitter release without full fusion of the vesicle with the plasma membrane.

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Figure 1: a, Arrangement of a CFE inside a patch pipette.
Figure 2: First exocytotic event shown in Fig. 1d on an expanded scale, showing amperometric current (a), imaginary part of the admittance (b, upper trace), and real part of admittance (b, lower trace) after baseline subtraction.
Figure 3: Event 6 shown in Fig. 1d on an expanded timescale, showing amperometric trace (a), imaginary part of admittance (b, upper trace), and real part of admittance (b, lower trace).

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Acknowledgements

We thank J. Pauli and the staff of the mechanical workshop for their contributions to the development and production of the adjustable pipette holder; J. Tritthard for the development of the amperometric amplifier; I. Wunderlich and R. Hinz-Herkommer for technical assistance; and E. Neher for his comments and encouragement. This work was supported by grants from the Deutsche Forschungsgemeinschaft to M.L. and from the Spanish Ministerio de Educación y Cultura (DGICYT) to G.A.T. A.A. was supported by an FPI fellowship from DGICYT.

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Author notes

  1. G. Dernick & M. Lindau

    Present address: School of Applied and Engineering Physics, Cornell University, Ithaca, New York, 14853-2501, USA

Authors and Affiliations

  1. Department of Molecular Cell Research, MPI f. Medical Research, D-69028, Heidelberg, Germany

    A. Albillos, G. Dernick, H. Horstmann, W. Almers & M. Lindau

  2. Department of Physiology & Biophysics, University of Seville, Sevilla, E-41009, Spain

    G. Alvarez de Toledo

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Correspondence to M. Lindau.

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Albillos, A., Dernick, G., Horstmann, H. et al. The exocytotic event in chromaffin cells revealed by patch amperometry. Nature 389, 509–512 (1997). https://doi.org/10.1038/39081

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