Murine Drinking Models in the Development of Pharmacotherapies for Alcoholism: Drinking in the Dark and Two-bottle Choice

Summary

Alcohol Use Disorder (AUD) is a major national health problem and the development of more effective treatments is required to offset the needs of this patient population. To this end, the following protocol utilizes two simple rodent drinking models to assess the preclinical efficacy of lead anti-alcohol compounds.

Abstract

Alcohol Use Disorder (AUD) is a major problem with more than an estimated 76 million people worldwide meeting the diagnostic criteria. Current treatments are limited to three FDA-approved medications that are largely ineffective even when combined with psychosocial intervention, as is evident by the high relapse rate. As such, the search for more novel treatments represents an important public health goal. To this end, the following protocol utilizes two simple rodent drinking models to assess the preclinical efficacy of lead anti-alcohol compounds: two-bottle choice (TBC) and drinking in the dark (DID). The former allows mice to voluntary drink in moderation while the latter induces mice to voluntary consume a large amount of alcohol in a short period that mimics binge drinking. The simple and high throughput nature of both of these paradigms allow for rapid screening of pharmacological agents or for identifying strains of mice that exhibit certain voluntary drinking behavior.

Introduction

For the past 25 plus years, significant effort has been put towards developing medications for the treatment of Alcohol Use Disorder (AUD)¹.Although many advances have been made, AUD still remains a major public health problem, affecting over 18 million Americans, and costing over $220 billion annually^2,3. Currently there are only three FDA-approved medications, disulfiram, naltrexone, and acamprosate, all of which have yielded inconsistent results in clinical trials and limited success even when combined with psychosocial intervention in the clinic settings⁴

Protocol

All procedures described here have been approved by the Institutional Animal Care and Use Committees of the University of Southern California Health Sciences Campus.

1. Experimental Setup and Assembly

1. Acquire all of the following supplies and chemicals before the start of the study: mice, cages/metal cage tops, bedding, food, water, ethanol, pipets, sipper tops, shrink wrap, utility knife, zip ties, tape, Bunsen burner, scale, headlamp.
2. Obtain C57BL/6J mice, either from a commercial source or an in-house colony, keeping in mind that mice can be group housed until time of testing.
   ​NOTE: The total number of....

Results

In the following representative investigations, social drinking was modeled using the two-bottle choice (TBC) paradigm. Briefly, mice had access to two bottles of solution, one of which contained water, and the other a 10% (v/v) ethanol solution. Subjects were subsequently split and evenly assigned to drug treatment groups, moxidectin (MOX) vs. saline control, so that each group would have averaged ethanol intake levels with the least amount of variation.

Discussion

Worldwide estimates indicate that as many as 76 million people meet the criteria to warrant a diagnosis for Alcohol Use Disorder (AUD). Unfortunately, pharmaceutical treatments currently available are largely ineffective and further development is necessary to offset the needs of this clinical population²⁰. To this end, the following protocol aims to facilitate this endeavor by exemplifying two of the most basic rodent drinking paradigms: two-bottle-choice (TBC) and drinking in the dark (DID). Bot.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
1 L  graduated cylinder	VWR	https://us.vwr.com/store/product/20935285/marisco-single-scale-cylinder-graduates-john-m-maris-co	To prepare ethanol solution.
1 L glass bottle	Pyrex (Fisher Scientific)	https://www.fishersci.com/shop/products/pyrex-reusable-media-storage-bottles-12/p-42752	To prepare ethanol solution.
100 mL graduated cylinder	Fisher Scientific	https://www.fishersci.com/shop/products/kimble-chase-kimax-class-a-to-contain-graduated-cylinders-8/p-4369311	To prepare ethanol solution.
Analox			One potential method of analyzing DID blood samples is by using the analox machine
ball-bearing sipper tubes	Ancare Corp.	http://www.ancare.com/products/watering-equipment/open-drinking-tubes/straight-tubes-ball-point	Length: 2.5 inches, Diameter: 5/16 inches, Model: TD100
C57BL/6J Mice	Jackson lab	https://www.jax.org/strain/000664	May also come from internal breeding colony
disposable serological pipets	VWR International (VWR)	https://us.vwr.com/store/product/4760455/vwr-disposable-serological-pipets-polystyrene-sterile-plugged	10 mL, 18 mL, or 25 mL
ethanol, pure, 190 proof (95%), USP, KOPTEC	Decon Labs (VWR)	https://us.vwr.com/store/product/4542412/ethanol-pure-190-proof-95-usp-koptec	---
heat gun	Master Appliances Corp.	http://www.masterappliance.com/master-heat-guns-kits/
heat shrink tubing	---	---	Diameter: 3/8 inches
industrial knife/blade	---	---	---
metal cage plate	---	---	Should be available through the university/institutional vivarium
mouse RO water	---	---	Should be available through the university/institutional vivarium
portable electronic scale	Ohaus (VWR)	https://us.vwr.com/store/product/4789377/portable-electronic-cs-series-scales-ohaus	---
red light headlamp	nyteBright (Amazon)	https://www.amazon.com/LED-Headlamp-Flashlight-Red-Light/dp/B00R0LMMF8/ref=sr_1_1?ie=UTF8&qid=1499591137&sr=8-1-spons&keywords=red+lamp+headlamp&psc=1	---
silicone stoppers	Fisher	---	---
thermometer	Fisher Scientific	https://www.fishersci.com/shop/products/fisher-scientific-hygro-thermometer-clock-large-display-2/p-4077232	---
weigh boat	VWR International (VWR)	https://us.vwr.com/store/product/16773534/vwr-pour-boat-weighing-dishes	The lid from a pipete tip box is an appropriate alternative