title: Three distinct modes of exocytosis revealed by amperometry in neuroendocrine cells.
abstract: Neurotransmission requires Ca(2+)-dependent release of secretory products through fusion pores that open and reclose (partial membrane distention) or open irreversibly (complete membrane distention). It has been challenging to distinguish between these release modes; however, in the work presented here, we were able to deduce different modes of depolarization-evoked exocytosis in neuroendocrine chromaffin and PC12 cells solely by analyzing amperometric recordings. After we determined the quantal size (Q), event half-width (t(50)), event amplitude (I(peak)), and event decay time constant (τ(decay)), we fitted scatter plots of log-transformed data with a mixture of one- and two-dimensional Gaussian distributions. Our analysis revealed three distinct and differently shaped clusters of secretory events, likely corresponding to different modes of exocytosis. Complete membrane distention, through fusion pores of widely varying conductances, accounted for 70% of the total amount of released catecholamine. Two different kinds of partial membrane distention (kiss-and-run and kiss-and-stay exocytosis), characterized by mode-specific fusion pores with unitary conductances, accounted for 20% and 10%, respectively. These results show that our novel one- and two-dimensional analysis of amperometric data reveals new release properties and enables one to distinguish at least three different modes of exocytosis solely by analyzing amperometric recordings.
authors.affiliation: Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: G Th H
authors.initials: GT
authors.lastname: van Kempen
authors.forename: H T
authors.initials: HT
authors.lastname: vanderLeest
authors.forename: R J
authors.initials: RJ
authors.lastname: van den Berg
authors.forename: P
authors.initials: P
authors.lastname: Eilers
authors.forename: R H S
authors.initials: RH
authors.lastname: Westerink
mesh_terms: D000200:Action Potentials / Q000502:physiology; D000818:Animals; D019439:Chromaffin Cells / Q000166:cytology / Q000378:metabolism; D055664:Electrochemical Techniques / Q000379:methods*; D005089:Exocytosis; D051379:Mice; D055099:Neuroendocrine Cells / Q000166:cytology* / Q000378:metabolism; D016011:Normal Distribution; D016716:PC12 Cells; D051381:Rats