Intravenous Self‐Administration of Ethanol in Mice

Nicholas J. Grahame1, Christopher L. Cunningham2

1 Indiana University/Purdue University at Indianapolis, Indianapolis, Indiana, 2 Oregon Health Science University, Portland, Oregon
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Unit 9.11
DOI:  10.1002/0471142301.ns0911s19
Online Posting Date:  August, 2002
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

A more complete understanding of alcohols reinforcing actions is obtained when multiple behavioral procedures are used, some of which bypass taste factors. This unit describes a method for assessing the reinforcing effects of alcohol in mice using the most widely accepted procedure for assessing drug reward: intravenous self‐administration.

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Basic Protocol 1: Intravenous Ethanol Self‐Administration in Mice
  • Commentary
  • Literature Cited
  • Figures
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

Basic Protocol 1: Intravenous Ethanol Self‐Administration in Mice

  Materials
  • 15‐ to 35‐g adult male or female mice, C57BL/6J or DBA/2J (The Jackson Laboratory) or other strains if desired
  • Mouse anesthetic cocktail: 17 mg/ml ketamine, 3 mg/ml acepromazine, 6 mg/ml xylazine
  • Low‐viscosity cyanoacrylic cement (e.g., Nexaband, Closure Medical) in a 1‐ml tuberculin syringe mated to an 18‐G needle
  • Heparinized saline solution for injection
  • Standard rodent diet
  • Sterile saline
  • Brevital sodium for injection
  • 95% to 100% ethanol diluted with 0.9% saline to produce a unit dose of 75 mg/kg body weight in 5 µl infusion volume (i.e., 28.3% to 75.6% v/v ethanol)
  • Isothermic heating pad
  • Scale accurate to 0.1 g
  • Rectal temperature probe, accurate to 0.1°C
  • Surgical scissors, scalpel, supplies (swabs, Betadine, hair clippers, 5‐0 surgical silk)
  • Dissection microscope with fiber‐optic light source
  • Fine‐tip forceps (e.g., Dumont no. 55, Fine Science Tools)
  • Subcutaneous stainless steel mouse saddles constructed of 27‐G hypodermic tubing and wire; 1 cm of tubing is exteriorized to provide an attachment point for drug‐delivery tubing, and comes with removable cap; can be purchased from IITC (http://www.iitcinc.com/). Prior to surgery, mate the exterior portion of the saddle to a 1‐ml tuberculin syringe filled with heparinized saline to permit delivery of anticoagulant during the surgery.
  • Caps for mouse saddles, constructed of PE‐50 tubing cinched closed at one end (using heat and a pair of forceps) to create a seal. Be sure to test caps for a water‐tight seal before use.
  • Cannulae composed of 50‐mm Silastic tubing (Dow Corning; 0.012‐in. i.d.; 0.025‐in. o.d.) with a movable 3‐mm cuff of PE‐50 threaded over the Silastic, ∼11 mm from the end of the catheter. The jugular end of the cannula should be beveled, and the catheter end secured to the mouse saddle with a 3‐mm section of heat‐shrink tubing.
  • 6 × 4‐in. piece of ½‐in. thick Plexiglas, with a 0.25‐in. slot running halfway down the long end
  • 50‐g weight
  • 27‐G, ½‐in. hypodermic needle, bent 120°, 5 mm from the beveled tip using pliers
  • Polycarbonate caging (27.9 × 9.5 × 12.7 cm) with corn cob or Cell‐sorb bedding
  • Drug delivery system (e.g., PHM‐100 syringe pump from Med Associates)
  • Restraint tube (e.g., IITC restrainer model no. 84)
  • Small hemostats (2‐3; e.g., Halsted‐Mosquito straight, from FST)
  • Four boxes with aluminum end walls and Plexiglas side walls (30 × 15 × 30 cm), with wire‐mesh floors, and a hinged, drilled (for air holes) Plexiglas cover
  • Sound‐attenuating cubicles (e.g., Med Associates, Env‐021M) to house aluminum/Plexiglas boxes
  • Two nosepoke response devices located at either end of the boxes in the aluminum walls
  • Optional: Locomotor activity can be measured via 6‐8 pairs of infrared photoemitter‐receiver diodes mounted at regular intervals on the Plexiglas walls, 2.5 cm above the floor (unit 8.1)
  • Leash for fluid delivery from swivel to mouse; this can be an aperture of tygon tubing that will fit both the fluid‐bearing swivel and the subcutaneous saddle
  • Fluid‐bearing swivel (e.g., Instech Laboratories; stainless steel 25‐G single channel)
  • Personal computer for event recording and response‐contingent light and drug delivery
NOTE: Sterilize the subcutaneous saddle and surgical instruments by soaking in sterilization solution (e.g., Cidex) ≥10 hr before implanting.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

  •   FigureFigure 9.11.1 Operant intravenous self‐administration apparatus used to deliver response‐contingent drug infusions and collect data during i.v. self‐administration sessions. Each nosepoke hole should have an aperture of 2.3 cm, and be mounted 1.5 cm above the wire mesh floor. A bulb (e.g., 5 VDC GE‐47 bulb) should be mounted inside the hole to provide indirect illumination upon completion of a reinforced response. Responses consist of occlusion of an infrared photoemitter‐receiver pair for at least 50 msec followed by nonocclusion; photoemitter/receivers are available at an electronics shop or from Med Associates.
  •   FigureFigure 9.11.2 A schematic of the right jugular incision site immediately following implantation of the catheter. The incision should be ∼1.5 cm2 once retracted at each corner. The catheter terminates at the dorsal subcutaneous saddle assembly, and should be filled with heparinized saline during and after surgery. The suture placed over the PE section of the catheter should be anchored into musculature immediately dorsal to the jugular, for added catheter stability. See text for further details.
  •   FigureFigure 9.11.3 Responding for a 75 mg/kg ethanol infusion on an FR‐3 schedule of reinforcement during 2‐hr daily sessions. Nosepoking was reinforced on one side of the chamber (“correct nosepokes”), and nonreinforced on the other. Beta‐endorphin‐deficient mice acquired responding while wild‐type mice did not. Open and closed squares indicate correct and incorrect nosepokes, respectively, for beta‐endorphin deficient mice, while asterisks and plus signs indicate correct and incorrect nosepokes for wild‐type mice. Beta‐endorphin‐deficient mice had both a higher rate of responding and a greater percentage of total responses on the correct side compared to wild‐type mice. Bars indicate standard errors; n = 14 − 17. Reprinted with permission from Grahame et al. ().

Videos

Literature Cited

Literature Cited
   Belknap, J.K., Belknap, N.D., Berg, J., and Coleman, R.R. 1977. Preabsorptive vs. postabsorptive control of ethanol intake in C57BL/6J and DBA/2J mice. Behav. Genet. 7:414‐425.
   Belknap, J.K., Coleman, R.R., and Foster, K. 1978. Alcohol consumption and sensory threshold differences between C57BL/6J and DBA/2J mice. Physiol. Psych. 6:71‐74.
   Collins, R.J., Weeks, J.R., Cooper, M.M., Good, P.I., and Russel, R.R. 1984 Prediction of abuse liability of drugs using iv self‐administration by rats. Psychopharmacology 82:6‐13.
   Cunningham, C.L. 1995. Localization of genes influencing ethanol‐induced conditioned place preference and locomotor activity in BXD recombinant inbred mice. Psychopharmacology 120:28‐41.
   Deneau, G., Yanagita, T., and Seevers, M.H. 1969. Self‐administration of psychoactive substances by the monkey. Psychopharmacologia 16:30‐48.
   Deroche, V., Caine, S.B., Heyser, C.J., Polis, I., Koob, G.F., and Gold, L.H. 1997. Differences in the liability to self‐administer intravenous cocaine between C57BL/6 × SJL and BALB/cByJ mice. Pharm. Biochem. Behav. 57:429‐440.
   Grahame, N.J. and Cunningham, C.L. 1995. Genetic differences in intravenous cocaine self‐administration between C57BL/6J and DBA/2J mice. Psychopharmacology 122:281‐291.
   Grahame, N.J. and Cunningham, C.L. 1997. Intravenous ethanol self‐administration in C57BL/6J and DBA/2J mice. Alcohol. Clin. Exp. Res. 21:56‐62.
   Grahame, N.J., Phillips, T.J., Burkhart‐Kasch, S., and Cunningham, C.L. 1995. Intravenous cocaine self‐administration in the C57BL/6J mouse. Pharm. Biochem. Behav. 51:827‐834.
   Grahame, N.J., Low, M.J., and Cunningham, C.L. 1998. Intravenous self‐administration of ethanol in beta‐endorphin deficient mice. Alcohol. Clin. Exp. Res. 22:1099‐1105.
   Kuzmin, A. and Johansson, B. 1999. Expression of c‐fos, NGFI‐A and secretogranin II mRNA in brain regions during initiation of cocaine self‐administration in mice. Eur. J. Neurosci. 11:3694‐3700.
   Martellotta, M.C., Cossu, G., Fattore, L., Gessa, G.L., and Fratta, W. 1998a. Intravenous self‐administration of gamma‐hydroxybutyric acid in drug‐naive mice. Eur. Neuropsychopharm. 8:293‐296.
   Martellotta, M.C., Cossu, G., Fattore, L., Gessa, G.L., and Fratta, W. 1998b. Self‐administration of the cannabinoid receptor agonist WIN 55,212‐2 in drug‐naive mice. Neuroscience 85:327‐330.
   McLearn, G.E. and Rodgers, D.A. 1959. Differences in alcohol preference among inbred strains of mice Quarterly Journal for the Study of Alcohol 20:691‐695.
   Piazza, P.V., Deminiere, J.M., Le Moal, M., and Simon, H. 1989. Factors that predict individual vulnerability to amphetamine self‐administration Science 245:1511‐1513.
   Roberts, A.J., Polis, I.Y., and Gold, L.H. 1997. Intravenous self‐administration of heroin, cocaine, and the combination in Balb/c mice Eur. J. Pharm. 326:119‐125.
   Rocha, B.A., Ator, R., Emmett‐Oglesby, M.W., and Hen, R. 1997. Intravenous cocaine self‐administration in mice lacking 5‐HT1B receptors. Pharm. Biochem. Behav. 57:407‐412.
   Stewart, R.B. and Grupp, L.A. 1992. Models of alcohol consumption using the laboratory rat. In Animal Models of Drug Addiction (A.A. Boulton, G.B. Baker, and P.H. Wu, eds.) pp. 5‐7. Humana Press, Totowa, N.J.
   Yokel, R.A. 1987. Intravenous self‐administration: Response rates, the effects of pharmacological challenges, and drug preferences. In Methods of Assessing the Reinforcing Properties of Abused Drugs (M.A. Bozarth, ed.) pp. 1‐34. Springer‐Verlag, New York.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library