Dear Afzal,

I do not see a clear reason why you cannot combine a 24-h sleep deprivation (SD) with longitudinal measurements of sucrose preference (SP) over a few days. You do a training phase over 3-5 days, then SD for the test group of mice, and then measure SP afterwards for both control and test groups.

During the period of SD I would supply both control and test groups with two bottles of water, no sucrose in.

How do you actually do SD (handling, automatically)?

You might also combine SPT with some other tests for depression/anxiety-like behavior (forced swimming, tail suspension, open field) to make your conclusions more robust.

Good luck!

Dear Alexei Leliavski , thanx for the detailed answer; We use automated treadmill method to induce SD. As most of the research show SPT training for 3-5 days and then water and food deprive animals for 12-24 hours before the test in SPT. However, if i do training for 3-5 days in SPT and then sleep deprive the animals for 24 h (during sleep deprivation procedure food and water is always available), so is it possible to water and food deprived the animals during 24 h Sleep deprivation and then test the mice in SPT?

I cannot comment whether yo should use water & food deprivation before SP testing (personally, I wouldn't, to keep it a bit more natural), but you surely should keep the same procedure for both SD and control group, the only difference will be the treadmill is either on or off. If possible, I would keep the mice in the cages with a treadmill already during habituation period. I'd give them a few days to get used to the cage environment and to the bottles. When I did SPT, I started the habituation phase with providing the mice two bottles with just water to let them get used to such system (4-5 days), and later put a 1% sucrose solution into one of them. I also swapped the bottles twice a day to avoid some preference to the bottle position (left/right) and kept the same bottles for a specific animal (smell is important for mice). It overall takes some efforts, of course, but reduces variability caused by these seemed-to-be-minor factors.

Thank you so much for your time, meanwhile i have gone through various reports that used SPT protocols.

Hello Afzal, some studies don't have training sessions for the SP, but if you think the treatment you are trying to assess is going to have a subtle impact, training would be advisable. If there is rationale to think that the treatment is going to have a strong impact, then testing without training could be enough. Some studies also do only 1, others 3 hours. You can add several protocols in your Ethics , so if you have any problems with your first choice, you can try others without the long wait for amendments...