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This video demonstrates a single-fiber recording of C-fiber nerve conduction in rats. An anesthetized rat is incised to expose the sciatic nerve, which provides sensory and motor supply to the lower limbs. The individual C-fibers are then exposed. Electrical stimulation is provided using stimulus electrodes implanted in the foot, and a response is recorded from a single nerve fiber.
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Animals
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Name | Company | Catalog Number | Comments |
Instruments and software used in single fiber recording | |||
Amplifier | Nihon kohden | MEZ-8201 | Amplification of the electrophysiological signals |
Bioelectric amplifier monitor | ShangHai JiaLong Teaching instrument factory | SZF-1 | Monitor firing process via sound which is transformed from physiological discharge signal |
Data acquisition and analysis system | CED | Spike-2 | Software for data acquisition and analysis |
Electrode manipulator | Narishige | SM-21 | Contro the movement of the electrode as required |
Hairspring tweezers | A.Dumont | 5# | Separate the single fiber |
Isolator | Nihon kohden | SS-220J | |
Memory oscilloscope | Nihon kohden | VC-9 | Display recorded discharge during |
experiment | |||
Stereomicroscope | ZEISS | SV-11 | Have clear observation when separate the local tissue and single fiber |
Stimulator | Nihon kohden | SEZ-7203 | Delivery of the electrical stimuli |
Von Frey Hair | Stoelting accompany | Delivery of the mechanical stimuli | |
Water bath | Scientz biotechnology Co., Ltd. | SC-15 | Heating paroline to maintain at 37oC |
Origin | Origin lab | Origin 8 | Software for drawing picture |
Other instruments | |||
Electronic balance | Sartorius | BS 124S | Weighing reagent |
pH Modulator | Denver Instrument | UB7 | Adjust pH to 7.4 |
Solutions/perfusion/chemicals | |||
Calcium chloride | Sigma-Aldrich | C5670 | Extracellular solution |
Chloralose | Shanghai Meryer Chemical Technology Co., Ltd. | M07752 | Mixed solution for Anesthesia |
D (+) Glucose | Sigma-Aldrich | G7528 | Extracellular solution |
Liquid Paraffin | TianJin HongYan Reagent Co., Ltd. | Maintain fiber wetting | |
Magnesium sulfate | Sigma-Aldrich | M7506 | Extracellular solution |
Potassium chloride | Sigma-Aldrich | P3911 | Extracellular solution |
Protease | Sigma-Aldrich | 62H0351 | Enzyme used for clearing the surface of DRG |
Sodium bicarbonate | Sigma-Aldrich | S5671 | Extracellular solution |
Sodium chloride | Sigma-Aldrich | S5886 | Extracellular solution |
Sodium phosphate monobasic | Sigma-Aldrich | S0751 | Extracellular solution |
Sucrose | Sigma-Aldrich | S0389 | Extracellular solution |
Urethane | Sigma-Aldrich | U2500 | Mixed solution for Anesthesia |
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Source: Mao, H., et al. Use of In Vivo Single-fiber Recording and Intact Dorsal Root Ganglion with Attached Sciatic Nerve to Examine the Mechanism of Conduction Failure. J. Vis. Exp. (2019).
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