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Exocytosis of single chromaffin granules in cell-free inside-out membrane patches.

Abstract：

:In chromaffin cells, exocytosis of single granules and properties of the fusion pore--the first connection between vesicular lumen and extracellular space --can be studied by cell-attached patch amperometry, which couples patch-clamp capacitance measurements with simultaneous amperometric recordings of transmitter release. Here we have studied exocytosis of single chromaffin granules and endocytosis of single vesicles in cell-free inside-out membrane patches by patch capacitance measurements and patch amperometry. We excised patches from chromaffin cells by using methods developed for studying properties of single ion channels. With low calcium concentrations in the pipette and bath, the patches showed no spontaneous exocytosis, but exocytosis could be induced in some patches by applying calcium to the cytoplasmic side of the patch. Exocytosis was also stimulated by calcium entry through the patch membrane. Initial conductances of the fusion pore were undistinguishable in cell-attached and excised patch recordings, but the subsequent pore expansion was slower in excised patches. The properties of exocytotic fusion pores in chromaffin cells are very similar to those observed in mast cells and granulocytes. Excised patches provide a tool with which to study the mechanisms of fusion pore formation and endocytosis in vitro.

journal_name

Nat Cell Biol

journal_title

Nature cell biology

authors

Dernick G,Alvarez de Toledo G,Lindau M

doi

10.1038/ncb956

keywords:

subject

Has Abstract

pub_date

2003-04-01 00:00:00

pages

358-62

issue

4

eissn

1465-7392

issn

1476-4679

pii

ncb956

journal_volume

5

pub_type

杂志文章

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