Electric Field Stimulation for the Functional Assessment of Isolated Dorsal Root Ganglion Neuron Excitability
- 21 January 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Annals of Biomedical Engineering
- Vol. 49 (3), 1110-1118
- https://doi.org/10.1007/s10439-021-02725-y
Abstract
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 Ca2+ 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 Ca2+ activity; half-maximal response voltage (EV50) increased from 13.4 V in untreated tissues to 21.2, 23.3, 51.5 (p < 0.05), and 60.6 V (p < 0.05) at 0.01, 0.1, 1, and 10 µM doses, respectively. This technique may help improve an understanding of neural signaling while retaining tissue structural organization and serves as a tool for the rapid ex vivo recording and assessment of neural activity.Keywords
Funding Information
- National Science Foundation (DGE-1745038)
- Orthopaedic Research and Education Foundation (P20-03413)
- National Institute of Arthritis and Musculoskeletal and Skin Diseases (AR070975, AR077678, AR069588, AR074441)
This publication has 41 references indexed in Scilit:
- Failure of action potential propagation in sensory neurons: mechanisms and loss of afferent filtering in C‐type units after painful nerve injuryJournal Of Physiology-London, 2013
- Imaging Calcium in NeuronsNeuron, 2012
- Miniaturized integration of a fluorescence microscopeNature Methods, 2011
- Development of Micro-Electrode Array Based Tests for Neurotoxicity: Assessment of Interlaboratory Reproducibility with Neuroactive ChemicalsFrontiers in Neuroengineering, 2011
- Temporal Control of Gene Deletion in Sensory Ganglia Using a Tamoxifen-Inducible Advillin-CreERT2 Recombinase MouseMolecular Pain, 2011
- Contribution of the activation of satellite glia in sensory ganglia to pathological painNeuroscience & Biobehavioral Reviews, 2009
- Neural plasticity after peripheral nerve injury and regenerationProgress in Neurobiology, 2007
- Neuronal somatic ATP release triggers neuron–satellite glial cell communication in dorsal root gangliaProceedings of the National Academy of Sciences of the United States of America, 2007
- Medium and large injured dorsal root ganglion cells increase TRPV-1, accompanied by increased α2C-adrenoceptor co-expression and functional inhibition by clonidinePain, 2005
- Tetrodotoxin Blockage of Sodium Conductance Increase in Lobster Giant AxonsThe Journal of general physiology, 1964