I'm construction mouse alcohol addiction model to study alcohol addiction and withdrawal symptoms, many papers said they used Intermittent Access to 20% Ethanol 2-Bottle Free-Choice (IA2BC) Drinking Procedure, which is to give mouse two bottles, one contains plain water, one contains 20% alcohol, after 24 hours, the alcohol bottle was replaced by water bottle for the next 24 hours. This pattern was repeated for 6-8 weeks.

The question is, can I use only one bottle that contains 10% alcohol as the only liquid source (as the concentration of 20% is too high)? Because using two-bottle free-choice is troublesome, more importantly, I'm not sure each mouse actually drinks alcohol, and even if it drinks, the amount may not be sufficient to induce any addiction or withdrawal behavior. Using one bottle can solve this problem, however, few articles claimed using only one bottle of alcohol.

And why many studies use two-bottle free choice and they claim this can induce depressive symptoms and anxiety after withdrawal. However, I didn't observe any anxiety symptoms after withdrawal using the open field test and elevated plus-maze in the last experiment.

Note: In my study, I performed cannular implantation on mice before behavior assay.