Abstract
The patch-clamp technique has been used to measure changes in membrane capacitance (Cm) of bovine lactotrophs in order to monitor fluctuations in cell surface area associated with exo- and endocytosis. Cells were prepared by an enrichment procedure and cultured for up to 14 d before use. Under whole-cell recording, cell cytoplasm was dialyzed with various Ca2(+)-containing solutions. The resting Cm of 6.05 +/- 1.68 pF was found to correlate well with squared cell radius, suggesting a specific Cm of 0.8 microF/cm2. Discrete Cm steps of 2-10 fF were recorded, which most likely reflect single fusion and retrieval events of prolactin-containing granules (0.2-0.6 microns in diameter). High Ca2+ resulted in a Cm increase of 20-50% from the resting value, demonstrating a role for [Ca2+]i in stimulus-secretion coupling. Spontaneous Cm changes have also been recorded, which presumably reflect prolactin secretion supported by a tonic influx of Ca2+ through the membrane. This is supported by the following findings: addition of Co2+ diminished or reversed the spontaneous Cm changes and decreased resting [Ca2+]i; and membrane depolarization increased Cm, indicating the role of voltage-activated channels in stimulus-secretion coupling. As bovine lactotrophs have been found to be largely devoid of spontaneous electrical activity, a mechanism involving modulation of a tonic Ca2+ influx is proposed; this is shown to provide adequate control of basal and triggered secretion monitored by Cm.
Dates
Type | When |
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Created | 21 years, 3 months ago (May 13, 2004, 7:48 p.m.) |
Deposited | 2 years, 1 month ago (July 27, 2023, 8:36 a.m.) |
Indexed | 11 months, 3 weeks ago (Sept. 8, 2024, 2:54 a.m.) |
Issued | 34 years, 6 months ago (March 1, 1991) |
Published | 34 years, 6 months ago (March 1, 1991) |
Published Online | 34 years, 6 months ago (March 1, 1991) |
Published Print | 34 years, 6 months ago (March 1, 1991) |
@article{Zorec_1991, title={Increased cytosolic calcium stimulates exocytosis in bovine lactotrophs. Direct evidence from changes in membrane capacitance.}, volume={97}, ISSN={1540-7748}, url={http://dx.doi.org/10.1085/jgp.97.3.473}, DOI={10.1085/jgp.97.3.473}, number={3}, journal={The Journal of general physiology}, publisher={Rockefeller University Press}, author={Zorec, R and Sikdar, S K and Mason, W T}, year={1991}, month=mar, pages={473–497} }