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The Journal of Neuroscience, January 14, 2004, 24(2):303-309; doi:10.1523/JNEUROSCI.1119-03.2004

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Cellular/Molecular
The Effects of Vesicular Volume on Secretion through the Fusion Pore in Exocytotic Release from PC12 Cells

L. A. Sombers,¹ H. J. Hanchar,¹ T. L. Colliver,^1,2 N. Wittenberg,¹ A. Cans,³ S. Arbault,⁴ C. Amatore,⁴ and A. G. Ewing^1,2

¹Department of Chemistry, 152 Davey Laboratory, The Pennsylvania State University, University Park, Pennsylvania 16802, ²Department of Neuroscience and Anatomy, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033, ³Department of Chemistry, Göteborg University, S-41296 Göteborg, Sweden, and ⁴Ecole Normale Superieure, Departement de Chimie, Unité Mixte de Recherche Centre National de la Recherche Scientifique 8640, 75231 Paris Cedex 05, France

Many spikes in amperometric records of exocytosis events initially exhibit a prespike feature, or foot, which represents a steady-state flux of neurotransmitter through a stable fusion pore spanning both the vesicle and plasma membranes and connecting the vesicle lumen to the extracellular fluid. Here, we present the first evidence indicating that vesicular volume before secretion is strongly correlated with the characteristics of amperometric foot events. L-3,4-Dihydroxyphenylalanine and reserpine have been used to increase and decrease, respectively, the volume of single pheochromocytoma cell vesicles. Amperometry and transmission electron microscopy have been used to determine that as vesicle size is decreased the frequency with which foot events are observed increases, the amount and duration of neurotransmitter released in the foot portion of the event decreases, and vesicles release a greater percentage of their total contents in the foot portion of the event. This previously unidentified correlation provides new insight into how vesicle volume can modulate the activity of the exocytotic fusion pore.

Key words: fusion pore; exocytosis; amperometry; transmission electron microscopy; PC12 cells; vesicle volume; foot

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Received April 15, 2003; revised September 15, 2003; accepted November 14, 2003.

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