Our research pertained the field of the alcohol using disorder. This study highlights the methodology commonly using in the established the Alco dependence mouse model. This model is a chronic intermittent ethanol vapor paired with the two bottle choice paradigm.
The CIE two bottle choice paradigm. Automated two bottle choice equipment allows for the monitoring of mouse drinking behavior by recording the frequency of super touches. This enables us to acquire additional information beyond the volume consumed, such as timing of drinking bouts throughout a drinking session.
In preclinical studies of the alcohol using disorder, the approaches such as the whole brain imaging, chemogenetics, and optogenetics with various of the viral transgenetic mice are often used to advance the research in this field. Other approaches such as the computational modeling are also be used. The two bottle choice CIE model for alcohol dependence has been established for many years.
However, we are hoping to make the adoption of the approach more readily accessible to the broader research community. Compare with the other techniques using to study the alcohol using disorder. The two bottle choice and the CIE paradigm has a several advantages that are increased the translational relevance, such as induction of the significant withdrawal symptoms and the development of the escalation of the alcohol intake.
This approach is ideal for its use to study the relevant neuro biological changes caused by the alcohol dependence. To begin habitation, move the group housed mice into their individual drinking cages 30 minutes before the lights turn off. Record the volume of each drinking bottle and then allow the mice to drink for two hours.
Record the volumes of the bottles after the drinking period. Now expose the mice to at least two weeks of baseline two bottle choice drinking to establish a stable ethanol intake. Test the mice by giving them a water bottle and a second bottle containing 15%ethanol.
Record the volumes of each bottle before and after the two hour drinking period. Once the mice have achieved a stable baseline of ethanol intake, divide them into two groups based on their intake and put them back in their home cage. Intraperitoneally inject the mice with an ethanol-pyrazole solution.
Inject all the air controls and the other groups with the same pyrazol dose mixed with saline. After injections, place the air control mice in their home cages and place the CIE cages inside the vapor chamber. Lock the doors and set the pump to the appropriate vapor level.
Next, set the runtime of the experiment and then press start on the CIE vapor chamber control screen 30 minutes before the vapor system turns off. Determine the blood ethanol concentration of the mice. After collecting the blood, return the mice to the two bottle choice testing the week after the CIE.
Centrifuge the blood samples at 208 G for 1.0 minutes at four degrees Celsius to separate the plasma. Use an oxygen rate ethanol analyzer to determine the blood ethanol concentration. The initial baseline ethanol intake during the three week period stabilized at 2.00 plus or minus 0.21 grams per kilogram before the CIE.
A Bonferroni post hoc test showed that the CIE mice increased their ethanol intake significantly at post vapor week six compared to their baseline drinking volume. The CIE mice consumed significantly more ethanol than the non-dependent mice at PV6. The Bonferroni post hoc test also showed that at PV6, the ethanol dependent mice had significantly higher ethanol intake compared to their own baseline and non-dependent mice.
The analyzed blood ethanol concentrations of the CIE group showed a weekly average of 173.12 milligrams per deciliter.
