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Voltage-sensitive dye recording of action potentials and synaptic potentials from sympathetic microcultures

Chien, C. B. and Pine, J. (1991) Voltage-sensitive dye recording of action potentials and synaptic potentials from sympathetic microcultures. Biophysical Journal, 60 (3). pp. 697-711. ISSN 0006-3495. PMCID PMC1260113. https://resolver.caltech.edu/CaltechAUTHORS:20170408-203353009

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Abstract

Given the appropriate multicell electrophysiological techniques, small networks of cultured neurons (microcultures) are well suited to long-term studies of synaptic plasticity. To this end, we have developed an apparatus for optical recording from cultured vertebrate neurons using voltage-sensitive fluorescent dyes (Chien, C.-B., and J. Pine. 1991. J. Neurosci. Methods. 38:93–105). We evaluate here the usefulness of this technique for recording action potentials and synaptic potentials in microcultures of neurons from the rat superior cervical ganglion (SCG). After extensive dye screening and optimization of conditions, we chose the styryl dye RH423, which gave fast linear fluorescence changes of approximately 1%/100 mV for typical recordings. The root mean square noise of the apparatus (limited by shot noise) was typically 0.03%, equivalent to 3 mV of membrane potential. Illumination for at least 100 flashes of 100 ms each caused no noticeable photodynamic damage. Our results show that voltage-sensitive dyes can be used to record from microcultures of vertebrate neurons with high sensitivity. Dye signals were detected from both cell bodies and neurites. Signals from presumptive dendrites showed hyperpolarizations and action potentials simultaneous with those in the cell body, while those from presumptive axons showed delayed propagating action potentials. Subthreshold synaptic potentials in the cell body were occasionally detectable optically; however, they were usually masked by signals from axons passing through the same pixel. This is due to the complex anatomy of SCG microcultures, which have many crisscrossing neurites that often pass over cell bodies. Given a simpler microculture system with fewer neurites, it should be possible to use dye recording to routinely measure subthreshold synaptic strengths.


Item Type:Article
Related URLs:
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http://dx.doi.org/10.1016/S0006-3495(91)82099-9DOIArticle
https://www.sciencedirect.com/science/article/pii/S0006349591820999PublisherArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1260113/PubMed CentralArticle
Additional Information:© 1991 The Biophysical Society. Published by Elsevier. Received for publication 17 January 1991 and in final form 2 May 1991. We thank Drs. Amiram Grinvald and Leslie Loew for providing dyes and expertise, Dr. Helen Rayburn for the initial dye screening, Becky Tanamachi for tissue culture work, and Dr. Wade Regehr for helpful comments on the manuscript. This work was supported by grants to Dr. Pine from the System Development Foundation and the National Institutes of Health (grant NS22450-02).
Funders:
Funding AgencyGrant Number
System Development FoundationUNSPECIFIED
NIHNS22450-02
Issue or Number:3
PubMed Central ID:PMC1260113
Record Number:CaltechAUTHORS:20170408-203353009
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20170408-203353009
Official Citation:C.B. Chien, J. Pine, Voltage-sensitive dye recording of action potentials and synaptic potentials from sympathetic microcultures, Biophysical Journal, Volume 60, Issue 3, 1991, Pages 697-711, ISSN 0006-3495, https://doi.org/10.1016/S0006-3495(91)82099-9. (http://www.sciencedirect.com/science/article/pii/S0006349591820999)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:76388
Collection:CaltechAUTHORS
Deposited By: 1Science Import
Deposited On:30 Mar 2018 22:51
Last Modified:03 Oct 2019 17:00

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