| Low-noise recording of single-vesicle capacitance steps in cell-attached patches. | |
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MedLine Citation:
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PMID: 18369954 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Capacitance recording provides a readout of membrane area that can be used to monitor exo- and endocytosis in neurons and secretory cells in real time. By interfacing a lock-in amplifier to a patch-clamp amplifier, the capacitance of cell-attached membrane patches can be measured with sufficient sensitivity to reveal the fusion and retrieval of single vesicles as unitary stepwise changes in capacitance. The small size of many secretory vesicles, especially of synaptic vesicles, places a premium on the reduction of noise in a capacitance recording. With care, the capacitance noise in cell-attached patches can be reduced to below 10 aF root-mean-square (rms), thus bringing into view steps resulting from the fusion of vesicles as small as about 18 nm in diameter. Thus, the lowest achievable noise level enables the resolution of changes in capacitance associated with the smallest secretory vesicles. This chapter presents the method of capacitance recording from cell-attached patches with an emphasis on noise reduction. It also addresses the closely related issue of extracting fusion pore properties from these recordings. |
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Authors:
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Vitaly Klyachko; Zhenjie Zhang; Meyer Jackson |
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Publication Detail:
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Type: In Vitro; Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Methods in molecular biology (Clifton, N.J.) Volume: 440 ISSN: 1064-3745 ISO Abbreviation: Methods Mol. Biol. Publication Date: 2008 |
Date Detail:
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Created Date: 2008-03-28 Completed Date: 2008-05-15 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9214969 Medline TA: Methods Mol Biol Country: United States |
Other Details:
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Languages: eng Pagination: 283-95 Citation Subset: IM |
Affiliation:
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Howard Hughes Medical Institute and Molecular Neurobiology Laboratory, The Salk Institute, La Jolla, CA, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Membrane / metabolism* Electric Capacitance Exocytosis* Membrane Fusion* Membrane Potentials Models, Biological Neuropeptides / metabolism* Patch-Clamp Techniques* Pituitary Gland, Posterior / metabolism* Rats Rats, Sprague-Dawley Secretory Vesicles / metabolism* Synaptic Vesicles / metabolism Time Factors |
| Grant Support | |
ID/Acronym/Agency:
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//Howard Hughes Medical Institute |
| Chemical | |
Reg. No./Substance:
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0/Neuropeptides |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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