Voltage-clamp and current-clamp recordings from mammalian DRG neurons

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Abstract

We provide here detailed electrophysiological protocols to study voltage-gated sodium channels and to investigate how wild-type and mutant channels influence firing properties of transfected mammalian dorsal root ganglion (DRG) neurons. Whole-cell voltage-clamp recordings permit us to analyze kinetic and voltage-dependence properties of ion channels and to determine the effect and mode of action of pharmaceuticals on specific channel isoforms. They also permit us to analyze the role of individual sodium channels and their mutant derivatives in regulating firing of DRG neurons. Five to ten cells can be recorded daily, depending on the extent of analysis that is required. Because of different internal solutions that are used in voltage-clamp and current-clamp recordings, only limited information can be obtained from recording the same neuron in both modes. These electrophysiological studies help to elucidate the role of specific channels in setting threshold and suprathreshold responses of neurons, under normal and pathological conditions.

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Figure 1: Common problems associated with voltage-clamp recordings of sodium currents in DRG neurons.
Figure 2: L858H mutation increases firing frequency in DRG and decreases firing frequency in SCG neurons.
Figure 3: A mutation in sodium channel Nav1.7 (F1449V), associated with the inherited pain condition erythromelalgia, lowers the current threshold for action potential generation and increases rate of repetitive firing of DRG neurons.

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Acknowledgements

We thank Dr. Jin Sung Choi, Dr. Xiaoyang Cheng, Lynda Tyrrell, Larry Macala, Shujun Liu, Rachel Blackman, Bart Toftness and other members of our groups for valuable assistance in development and refinement of the techniques described in this article. Work in S.G.W. Laboratory is supported in part by grants from the National Multiple Sclerosis Society and the Rehabilitation Research and Development Service and Medical Research Service, Department of Veterans Affairs. T.R.C. was supported by research grant NS053422 from the National Institutes of Health. The Center for Neuroscience and Regeneration Research is a Collaboration of the Paralyzed Veterans of America and the United Spinal Association with Yale University.

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Correspondence to Stephen G Waxman.

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Cummins, T., Rush, A., Estacion, M. et al. Voltage-clamp and current-clamp recordings from mammalian DRG neurons. Nat Protoc 4, 1103–1112 (2009) doi:10.1038/nprot.2009.91

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