Sampling Phasic Dopamine Signaling with Fast‐Scan Cyclic Voltammetry in Awake, Behaving Rats

S.M. Fortin1, J.J. Cone1, S. Ng‐Evans2, J.E. McCutcheon3, M.F. Roitman4

1 Graduate Program in Neuroscience, University of Illinois at Chicago, Chicago, Illinois, 2 Department of Psychiatry and Behavioral Sciences, University of Washington, Seattle, Washington, 3 Department of Cell Physiology & Pharmacology, University of Leicester, Leicester, United Kingdom, 4 Department of Psychology, University of Illinois at Chicago, Chicago, Illinois
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Unit 7.25
DOI:  10.1002/0471142301.ns0725s70
Online Posting Date:  January, 2015
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Abstract

Fast‐scan cyclic voltammetry (FSCV) is an electrochemical technique that permits the in vivo measurement of extracellular fluctuations in multiple chemical species. The technique is frequently utilized to sample sub‐second (phasic) concentration changes of the neurotransmitter dopamine in awake and behaving rats. Phasic dopamine signaling is implicated in reinforcement, goal‐directed behavior, and locomotion, and FSCV has been used to investigate how rapid changes in striatal dopamine concentration contribute to these and other behaviors. This unit describes the instrumentation and construction, implantation, and use of components required to sample and analyze dopamine concentration changes in awake rats with FSCV. © 2015 by John Wiley & Sons, Inc.

Keywords: fast‐scan cyclic voltammetry; dopamine; reward; nucleus accumbens; motivation; reinforcement

     
 
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Table of Contents

  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1:

  Materials
  • Isopropyl alcohol (Sigma‐Aldrich, cat. no. I9030), filtered
  • Devcon 5‐min epoxy (Fisher Scientific, cat. no. NC9987160)
  • 1 N HCl (Fisher Scientific, cat. no. SA48‐1)
  • Sprague Dawley Rats (Charles River)
  • Ketamine
  • Xylazine
  • Betadine
  • Ethanol
  • Jet‐repair dental acrylic (Lang Dental, cat. no. 1405)
  • Carbon fiber (7 μm in diameter; Goodfellow Corp., cat. no. C005722)
  • 10‐cm borosilicate glass capillary tube (4 in., 0.6 mm OD × 0.4 mm ID; A‐M Systems, cat. no. 624500)
  • Thermo/Barnant vacuum pump (Midland Scientific, cat. no. BAR 400‐3910)
  • Vertical microelectrode puller (Narishige Group, cat. no. PE‐22)
  • Scissors
  • Scalpel with a #11 surgical blade (Fisher Scientific, cat. no. 14‐840‐16)
  • Light microscope with a graticule in the eyepiece
  • Microscope slide coated in clear packing tape
  • Sculpting putty
  • Gold pins (Newark, cat. no. SPC15509)
  • Bare wires (30‐G, 3 in. (7.6 cm), UL1423 type, Squires Electronics, Cornelius, or custom)
  • Silverprint conductive paint (GC Electronics, cat. no. 22‐024)
  • Three types of heat‐shrinkable flexible polyolefin tubing: 3/32 in. i.d. (APD, cat. no. 00‐051128‐59888‐7), 3/64 in. i.d., clear (Digi‐Key, cat. no. KY364C‐ND), and 3/64 in. i.d. (Altex, cat. no. HST3/64)
  • Micromanipulator (University of Illinois at Chicago Research Resources Center, custom; Fig. A), loaded with a carbon fiber electrode (Fig. ).
  • Silver wire (0.5 mm diameter, Sigma‐Aldrich, cat. no. 327026‐20 G)
  • 9 V battery with stainless steel or copper wires soldered to the terminals
  • Twisted stainless‐steel stimulating electrodes (20 mm, Plastics One, cat. no. MS303T/2‐B/SPC)
  • Animal clippers
  • Small animal stereotaxic instrument (David Kopf Instruments, model 900) with:
    • Non‐rupture ear bars (David Kopf Instruments, cat. no. 955)
    • Clamp rod for stereotaxic frame (BASi, cat. no. MD‐1521)
    • Probe clamp (BASi, cat. no. MD‐1520)
  • Gauze
  • Stainless steel surgical screws (3/16 in., Small Parts through Amazon, ASIN, cat. no. B000FN5XE0)
  • Surgical drill bit (0.7 mm diameter tip, Fine Science Tools, cat. no. 19008‐07)
  • Guide cannula (BASi, cat. no. MD‐2251)
  • Reference electrode: polyimide‐insulated stainless steel electrode (0.01 in. diameter, 3 in. length, tapered tip size 8°, AC resistance 12 MΩ; A‐M Systems, cat. no. 571500)
  • Flow injection system (University of Illinois at Chicago Research Resources Center, custom)
  • Polyimide‐insulated stainless steel electrode (0.01 in. diameter, 3 in. length, tapered tip size 8°, AC resistance 12 MΩ; A‐M Systems, cat. no. 571500)
  • Lesion‐making device (Ugo Basile, cat. no. 53500)
  • Electric industrial heat gun (Weller, cat. no. 6966C)
  • Hardware:
    • Multifunction input/output card: PCI‐6052E (16 bit, 333 kHz) (National Instruments)
    • Potentiostat: EI‐400 biopotentiostat (Cypress Systems, cat. no. 66‐El‐400) or Custom (FSCV interface with integrated timing circuitry combined with amplifier/current‐to‐voltage converter headstage [Electronics and Materials Engineering (EME) Shop, Seattle, Washington]
    • Headstage: low‐pass‐filtered amplifier/current‐to‐voltage headstage, standard amplification is 200 nA/V. Up to four channels for FSCV. Various configurations of this headstage allow for recording in small animals (mice, birds, fish) and larger animals such as rats and monkeys (EME Shop, Seattle, Washington; see Fig. B‐C)
    • Commutator: 9‐ or 25‐channel (Crist Instruments, cat. no. 4‐TBC‐9 S or 4‐TBC‐25); a 25‐channel commutator with a liquid tube cannula is also available from Crist Instruments for experiments that utilize a fluid line (cat. no. 4‐TBC‐9‐LT) (Fig. B,C)
    • Breakout box interface and power supply: a multifunction piece of equipment that separates digital and analog signals, sets the grounding scheme, reduces environmental noise, and filters and divides the applied waveform [custom; Electronics and Materials Engineering (EME) Shop, Seattle, Washington; Fig. A]
    • Extra‐tall behavioral chamber with TTL output card (DIG‐726TTL) to allow for time‐stamping animal‐ and machine‐generated events in the voltammetry record (Med Associates, custom to accommodate experimental needs); the extra‐tall chamber diminishes the probability of artifacts imparted to the recording because of the manipulator housing the electrode knocking into the roof or walls of the chamber (Fig. C)
    • Computer with two, preferably three, full‐height and full‐length PCI slots
  • Hardware for stimulation:
    • Digital‐to‐analog card: PCI‐6711 (National Instruments, cat. no. 777740‐01); this card is used in conjunction with the PCI‐6052E (see Hardware, above) to reduce overlap of stimulation with the data acquisition scans
    • Constant‐current stimulator: e.g., Neurolog NL800 (Harvard Apparatus, cat. no. 650276), ISO‐flex stimulus isolator (A.M.P.I., Jerusalem, Israel) or Model 2200 analog stimulus isolator (A‐M Systems, cat. no. 850000) (Fig. A)
  • Software:
    • Analysis software options include: HDCV (Bucher et al., ); TarHeel CV (UNC Electronics Facility); or Demon (Yorgason et al., ; Wake Forest Innovations) for: visualization of color plots and current traces; cutting and splicing voltammetry files to TTLs (recorded events); performing principal components analysis (chemometrics); and compiling chemometric data files for graphing and statistical analysis
  • Additional reagents and equipment for injection of rodents (appendix 4f)
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Figures

Videos

Literature Cited

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