title: High-resolution membrane capacitance measurements for the study of exocytosis and endocytosis.
abstract: In order to understand exocytosis and endocytosis, it is necessary to study these processes directly. An elegant way to do this is by measuring plasma membrane capacitance (C(m)), a parameter proportional to cell surface area, the fluctuations of which are due to fusion and fission of secretory and other vesicles. Here we describe protocols that enable high-resolution C(m) measurements in macroscopic and microscopic modes. Macroscopic mode, performed in whole-cell configuration, is used for measuring bulk C(m) changes in the entire membrane area, and it enables the introduction of exocytosis stimulators or inhibitors into the cytosol through the patch pipette. Microscopic mode, performed in cell-attached configuration, enables measurements of C(m) with attofarad resolution and allows characterization of fusion pore properties. Although we usually apply these protocols to primary pituitary cells and astrocytes, they can be adapted and used for other cell types. After initial hardware setup and culture preparation, several C(m) measurements can be performed daily.
authors.affiliation: Laboratory of Neuroendocrinology-Molecular Cell Physiology, Institute of Pathophysiology, Faculty of Medicine, Ljubljana, Slovenia.
authors.forename: Boštjan
authors.initials: B
authors.lastname: Rituper
authors.forename: Alenka
authors.initials: A
authors.lastname: Guček
authors.forename: Jernej
authors.initials: J
authors.lastname: Jorgačevski
authors.forename: Ajda
authors.initials: A
authors.lastname: Flašker
authors.forename: Marko
authors.initials: M
authors.lastname: Kreft
authors.forename: Robert
authors.initials: R
authors.lastname: Zorec
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000737:chemistry*; D025802:Electric Capacitance; D004705:Endocytosis / Q000502:physiology*; D005089:Exocytosis / Q000502:physiology*; D008297:Male; D018408:Patch-Clamp Techniques; D051381:Rats; D017208:Rats, Wistar