Download Biomembranes Part L: Membrane Biogenesis: Processing and by Nathan P. Kaplan, Nathan P. Colowick, Sidney Fleischer, PDF

By Nathan P. Kaplan, Nathan P. Colowick, Sidney Fleischer, Becca Fleischer

FROM THE PREFACE: Volumes ninety six to ninety eight, elements J, okay, and L of the Biomembranes sequence, specialize in method to review membrane biogenesis, meeting, focusing on, and recycling. This box is without doubt one of the very fascinating and lively components of analysis. destiny volumes will take care of delivery and different points of membrane functionality

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Extra resources for Biomembranes Part L: Membrane Biogenesis: Processing and Recycling

Example text

When a cell, which may be considered as a hollow thin-walled sphere, is exposed to such an electric field, a voltage will be imposed across its membrane. The magnitude of this potential depends on the intensity of the field [E (V/cm)] and the size of the cell [radius r (cm)] and will vary around the cell's circumference. With reference to Fig. I the voltage at point P (Vp) across the membrane of a spherical cell is given by the simplified equation Vp = C" r" E " cos 0 volts where the value of the constant C depends on the relative conductivities of the extracellular fluid, the cytosol, and the membrane.

W 3-J 2- Z rim 1 - j J J c ~ J f t d J u3 0 0 I 2 I 5 FIELD I 10 STRENGTH (KV I 15 CM -1) FIG. 2. The expected membranepotential [from Eq. 2 #m diameteras a resultof exposureto variousfield strengths. Methodology Two methods have been used routinely to subject isolated cells to electric fields: (a) passing cells through a Coulter counter 2 and (b) discharging a capacitor through a cell suspension. The main disadvantage of using the Coulter-counter method is the length of time taken to harvest a worthwhile number of "leaky" cells, and for this reason we routinely use the second method, which is described below.

Temperature 37*. (c) Effect of Mg acetate on Ca-sensitive release of catecholamine. Cells on a K glutamate solution (Table I) were rendered "leaky" by 10 exposures of 2 kV cm -t (~ = 200 #sec) and immediately transferred to similar solutions containing various concentrations of free Mg 2+ (Mg acetate used). After 5-min incubation the cells were challenged with 10 m M Ca EGTA buffer and the catecholamine was determined 10 rain later. Temperature 37*. (d) Ca activation curves at different chloride concentrations.

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