G-Protein Modulation of Voltage-Gated Ca2+ Channels from Isolated Adult Rat Superior Cervical Ganglion Neurons.
Van B LuStephen R IkedaPublished in: Cold Spring Harbor protocols (2016)
Sympathetic neurons isolated from adult rat superior cervical ganglia (SCG) are a well-established model to study G-protein modulation of voltage-gated Ca(2+) channels (VGCCs). SCG neurons can be easily dissociated and are amendable to heterologous expression of genes, including genetic tools to study G-protein signaling pathways, within a time frame to maintain good spatial voltage-clamp control of membrane potential during electrophysiological recordings (8-36 h postdissociation). This protocol focuses on examining G-protein modulation of VGCCs; however, the procedures and experimental setup for acute application of agonists can be applied to study modulation of other ion channels (e.g., M-current, G-protein-coupled inwardly rectifying K(+) channels). We also discuss some common sources of artifacts that can arise during acute drug application onto dissociated neurons, which can mislead interpretation of results.
Keyphrases
- spinal cord
- liver failure
- randomized controlled trial
- oxidative stress
- genome wide
- signaling pathway
- gene expression
- dna methylation
- drug induced
- intensive care unit
- climate change
- long non coding rna
- computed tomography
- epithelial mesenchymal transition
- extracorporeal membrane oxygenation
- induced apoptosis
- mechanical ventilation
- genome wide identification
- image quality