Electric Field Stimulation for the Functional Assessment of Isolated Dorsal Root Ganglion Neuron Excitability
| Autor: | Marcos N. Barcellona, Ian M. Berke, Simon Y. Tang, Matthew G. Gayoso, Tom M. McGrath, Yu-Qing Cao, Munish C. Gupta, Chang Gui, Lori A. Setton, J Jordan Stivers |
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| Rok vydání: | 2021 |
| Předmět: |
Nervous system
0206 medical engineering Biomedical Engineering Mice Transgenic Tetrodotoxin Voltage-Gated Sodium Channels 02 engineering and technology Biology Article chemistry.chemical_compound Calcium imaging Dorsal root ganglion In vivo Ganglia Spinal medicine Animals Neurons Voltage-Gated Sodium Channel Blockers Microscopy Confocal 020601 biomedical engineering Electric Stimulation medicine.anatomical_structure chemistry GCaMP Calcium Neuron Neuroscience Ex vivo |
| Zdroj: | Ann Biomed Eng |
| ISSN: | 1573-9686 0090-6964 |
| DOI: | 10.1007/s10439-021-02725-y |
| Popis: | 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 < 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. |
| Databáze: | OpenAIRE |
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