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Measuring Action Potential Propagation Velocity in Murine Cortical Axons

Measuring the action potential (AP) propagation velocity in axons is critical for understanding neuronal computation. This protocol describes the measurement of propagation velocity using a combination of somatic whole cell and axonal loose patch recordings in brain slice preparations. The axons of...

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Autores principales: Kotler, Oron, Khrapunsky, Yana, Fleidervish, Ilya
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Bio-Protocol 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10632166/
https://www.ncbi.nlm.nih.gov/pubmed/37969753
http://dx.doi.org/10.21769/BioProtoc.4876
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author Kotler, Oron
Khrapunsky, Yana
Fleidervish, Ilya
author_facet Kotler, Oron
Khrapunsky, Yana
Fleidervish, Ilya
author_sort Kotler, Oron
collection PubMed
description Measuring the action potential (AP) propagation velocity in axons is critical for understanding neuronal computation. This protocol describes the measurement of propagation velocity using a combination of somatic whole cell and axonal loose patch recordings in brain slice preparations. The axons of neurons filled with fluorescent dye via somatic whole-cell pipette can be targeted under direct optical control using the fluorophore-filled pipette. The propagation delays between the soma and 5–7 axonal locations can be obtained by analyzing the ensemble averages of 500–600 sweeps of somatic APs aligned at times of maximal rate-of-rise (dV/dtmax) and axonal action currents from these locations. By plotting the propagation delays against the distance, the location of the AP initiation zone becomes evident as the site exhibiting the greatest delay relative to the soma. Performing linear fitting of the delays obtained from sites both proximal and distal from the trigger zone allows the determination of the velocities of AP backward and forward propagation, respectively. Key features • Ultra-thin axons in cortical slices are targeted under direct optical control using the SBFI-filled pipette. • Dual somatic whole cell and axonal loose patch recordings from 5–7 axonal locations. • Ensemble averaging of 500–600 sweeps of somatic APs and axonal action currents. • Plotting the propagation delays against the distance enables the determination of the trigger zone's position and velocities of AP backward and forward propagation.
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spelling pubmed-106321662023-11-15 Measuring Action Potential Propagation Velocity in Murine Cortical Axons Kotler, Oron Khrapunsky, Yana Fleidervish, Ilya Bio Protoc Methods Article Measuring the action potential (AP) propagation velocity in axons is critical for understanding neuronal computation. This protocol describes the measurement of propagation velocity using a combination of somatic whole cell and axonal loose patch recordings in brain slice preparations. The axons of neurons filled with fluorescent dye via somatic whole-cell pipette can be targeted under direct optical control using the fluorophore-filled pipette. The propagation delays between the soma and 5–7 axonal locations can be obtained by analyzing the ensemble averages of 500–600 sweeps of somatic APs aligned at times of maximal rate-of-rise (dV/dtmax) and axonal action currents from these locations. By plotting the propagation delays against the distance, the location of the AP initiation zone becomes evident as the site exhibiting the greatest delay relative to the soma. Performing linear fitting of the delays obtained from sites both proximal and distal from the trigger zone allows the determination of the velocities of AP backward and forward propagation, respectively. Key features • Ultra-thin axons in cortical slices are targeted under direct optical control using the SBFI-filled pipette. • Dual somatic whole cell and axonal loose patch recordings from 5–7 axonal locations. • Ensemble averaging of 500–600 sweeps of somatic APs and axonal action currents. • Plotting the propagation delays against the distance enables the determination of the trigger zone's position and velocities of AP backward and forward propagation. Bio-Protocol 2023-11-05 /pmc/articles/PMC10632166/ /pubmed/37969753 http://dx.doi.org/10.21769/BioProtoc.4876 Text en ©Copyright : © 2023 The Authors; This is an open access article under the CC BY license https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Methods Article
Kotler, Oron
Khrapunsky, Yana
Fleidervish, Ilya
Measuring Action Potential Propagation Velocity in Murine Cortical Axons
title Measuring Action Potential Propagation Velocity in Murine Cortical Axons
title_full Measuring Action Potential Propagation Velocity in Murine Cortical Axons
title_fullStr Measuring Action Potential Propagation Velocity in Murine Cortical Axons
title_full_unstemmed Measuring Action Potential Propagation Velocity in Murine Cortical Axons
title_short Measuring Action Potential Propagation Velocity in Murine Cortical Axons
title_sort measuring action potential propagation velocity in murine cortical axons
topic Methods Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10632166/
https://www.ncbi.nlm.nih.gov/pubmed/37969753
http://dx.doi.org/10.21769/BioProtoc.4876
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