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High-resolution membrane capacitance measurements for studying endocytosis and exocytosis in yeast.

Carrillo, Lucia and Cucu, Bayram and Bandmann, Vera and Homann, Ulrike and Hertel, Brigitte and Hillmer, Stefan and Thiel, Gerhard and Bertl, Adam (2015):
High-resolution membrane capacitance measurements for studying endocytosis and exocytosis in yeast.
In: Traffic (Copenhagen, Denmark), pp. 760-772, 16, (7), ISSN 1398-9219, [Article]

Abstract

Fusion of exocytotic vesicles with the plasma membrane gives rise to an increase in membrane surface area, whereas the surface area is decreased, when vesicles are internalized during endocytosis. Changes in membrane surface area, resulting from fusion and fission of membrane vesicles, can be followed by monitoring the corresponding proportional changes in membrane capacitance. Using the cell-attached configuration of the patch-clamp techniques, we were able to resolve the elementary processes of endocytosis and exocytosis in yeast protoplasts at high temporal and spatial resolution. Spontaneous capacitance changes were predominantly in the range of 0.2-1 fF, which translates to vesicles diameters of 90-200 nm. The size distribution revealed that endocytotic vesicles with a median at about 132 nm were smaller than exocytotic vesicles with a median at 155 nm. In energized and metabolizing protoplasts, endocytotic and exocytotic events occurred at frequencies of 1.6 and 2.7 events per minute, respectively. Even though these numbers appear very low, they are in good agreement with the observed growth rate of yeast cells and protoplasts.

Item Type: Article
Erschienen: 2015
Creators: Carrillo, Lucia and Cucu, Bayram and Bandmann, Vera and Homann, Ulrike and Hertel, Brigitte and Hillmer, Stefan and Thiel, Gerhard and Bertl, Adam
Title: High-resolution membrane capacitance measurements for studying endocytosis and exocytosis in yeast.
Language: English
Abstract:

Fusion of exocytotic vesicles with the plasma membrane gives rise to an increase in membrane surface area, whereas the surface area is decreased, when vesicles are internalized during endocytosis. Changes in membrane surface area, resulting from fusion and fission of membrane vesicles, can be followed by monitoring the corresponding proportional changes in membrane capacitance. Using the cell-attached configuration of the patch-clamp techniques, we were able to resolve the elementary processes of endocytosis and exocytosis in yeast protoplasts at high temporal and spatial resolution. Spontaneous capacitance changes were predominantly in the range of 0.2-1 fF, which translates to vesicles diameters of 90-200 nm. The size distribution revealed that endocytotic vesicles with a median at about 132 nm were smaller than exocytotic vesicles with a median at 155 nm. In energized and metabolizing protoplasts, endocytotic and exocytotic events occurred at frequencies of 1.6 and 2.7 events per minute, respectively. Even though these numbers appear very low, they are in good agreement with the observed growth rate of yeast cells and protoplasts.

Journal or Publication Title: Traffic (Copenhagen, Denmark)
Volume: 16
Number: 7
Divisions: 10 Department of Biology
10 Department of Biology > Yeast Membrane Biology
10 Department of Biology > Plant Membrane Biophysics
10 Department of Biology > Plant Cell Biology
Date Deposited: 03 Mar 2015 10:16
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