Abstract
Patch clamp is an electrophysiological technique that allows to analyze the activity of ion channels in neurons. In this chapter, we provide a detailed description of patch clamp protocol to measure the effect of a μ-opioid receptor agonist on the activity of G protein-coupled inwardly rectifying potassium (GIRK or Kir3) channels. This is performed in peripheral sensory neurons isolated from dorsal root ganglia (DRG) of mice without or with a chronic constriction injury (CCI) of the sciatic nerve, which models neuropathic pain. We describe the induction of the CCI , isolation and culture of DRG neurons, performance of the patch clamp recordings, and identification of opioid-responding neurons.
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References
Neher E, Sakmann B (1976) Single-channel currents recorded from membrane of denervated frog muscle fibres. Nature 260:799–802
Kornreich BG (2007) The patch clamp technique: principles and technical considerations. J Vet Cardiol 9:25–37
Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch 391:85–100
Nowotny T, Marra V (2013) Patch clamp technique. In: Jaeger D, Jung R (eds) Encyclopedia of computational neuroscience. Springer New York, New York, NY, pp 1–4
Machelska H, Celik MO (2018) Advances in achieving opioid analgesia without side effects. Front Pharmacol 9:1388
Gao XF, Zhang HL, You ZD, Lu CL, He C (2007) G protein-coupled inwardly rectifying potassium channels in dorsal root ganglion neurons. Acta Pharmacol Sin 28:185–190
Gorham L, Just S, Doods H (2014) Somatostatin 4 receptor activation modulates G-protein coupled inward rectifying potassium channels and voltage stimulated calcium signals in dorsal root ganglion neurons. Eur J Pharmacol 736:101–106
Spahn V, Nockemann D, Machelska H (2015) Electrophysiological patch clamp assay to monitor the action of opioid receptors. Methods Mol Biol 1230:197–211
Richner M, Jager SB, Siupka P, Vaegter CB (2017) Hydraulic extrusion of the spinal cord and isolation of dorsal root ganglia in rodents. J Vis Exp 119:55226
Nockemann D, Rouault M, Labuz D, Hublitz P, McKnelly K, Reis FC, Stein C, Heppenstall PA (2013) The K(+) channel GIRK2 is both necessary and sufficient for peripheral opioid-mediated analgesia. EMBO Mol Med 5:1263–1277
Stötzner P, Spahn V, Celik MO, Labuz D, Machelska H (2018) Mu-opioid receptor agonist induces Kir3 currents in mouse peripheral sensory neurons—effects of nerve injury. Front Pharmacol 9:1478
Rigaud M, Gemes G, Barabas ME, Chernoff DI, Abram SE, Stucky CL, Hogan QH (2008) Species and strain differences in rodent sciatic nerve anatomy: implications for studies of neuropathic pain. Pain 136:188–201
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Seitz, V., Stötzner, P., Labuz, D., Machelska, H. (2021). Patch Clamp Analysis of Opioid-Induced Kir3 Currents in Mouse Peripheral Sensory Neurons Following Nerve Injury. In: Spampinato, S.M. (eds) Opioid Receptors. Methods in Molecular Biology, vol 2201. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0884-5_12
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DOI: https://doi.org/10.1007/978-1-0716-0884-5_12
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-0884-5
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