Electric Field Stimulation for the Functional Assessment of Isolated Dorsal Root Ganglion Neuron Excitability

Genetically encoded calcium indicators have proven useful for characterizing dorsal root ganglion neuron excitability in vivo . Challenges persist in achieving high spatial–temporal resolutions in vivo , however, due to deep tissue imaging and motion artifacts that may be limiting technical factors...

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Bibliographic Details
Published in:Annals of biomedical engineering 2021-03, Vol.49 (3), p.1110-1118
Main Authors: Berke, Ian M., McGrath, Tom M., Stivers, J. Jordan, Gui, Chang, Barcellona, Marcos N., Gayoso, Matthew G., Tang, Simon Y., Cao, Yu-Qing, Gupta, Munish C., Setton, Lori A.
Format: Article
Language:English
Subjects:
Animals
Biochemistry
Biological and Medical Physics
Biomedical and Life Sciences
Biomedical Engineering and Bioengineering
Biomedicine
Biophysics
Calcium
Calcium - physiology
Calcium channels (voltage-gated)
Calcium ions
Classical Mechanics
Dorsal root ganglia
Drug screening
Electric fields
Electric Stimulation
Excitability
Ganglia, Spinal - physiology
Genetic code
Mice, Transgenic
Microscopy, Confocal
Neurons
Neurons - physiology
Original Article
Perturbation
Sodium channels (voltage-gated)
Stimulation
Tetrodotoxin
Tetrodotoxin - pharmacology
Voltage-Gated Sodium Channel Blockers - pharmacology
Voltage-Gated Sodium Channels - physiology
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Summary:Genetically encoded calcium indicators have proven useful for characterizing dorsal root ganglion neuron excitability in vivo . Challenges persist in achieving high spatial–temporal resolutions in vivo , however, due to deep tissue imaging and motion artifacts that may be limiting technical factors in obtaining measurements. Here we report an ex vivo imaging method, using a peripheral neuron-specific Advillin-GCaMP mouse line and electric field stimulation of dorsal root ganglion tissues, to assess the sensitivity of neurons en bloc . The described method rapidly characterizes Ca 2+ activity in hundreds of dorsal root ganglion neurons (221 ± 64 per dorsal root ganglion) with minimal perturbation to the in situ soma environment. We further validate the method for use as a drug screening platform with the voltage-gated sodium channel inhibitor, tetrodotoxin. Drug treatment led to decreased evoked Ca 2+ activity; half-maximal response voltage (EV 50 ) increased from 13.4 V in untreated tissues to 21.2, 23.3, 51.5 ( p  
ISSN:0090-6964
1573-9686
1573-9686
DOI:10.1007/s10439-021-02725-y