Addicted brain mapping and imaging using light-sheet fluorescence microscopy

Addiction is a disease that affects circuits and brain areas involved in reward, stress, and self‑control. Continued substance abuse can lead to dependence that is associated with withdrawal symptoms when drug availability is ceased, and increased intake upon relapse. With many brain structures involved in the disease, the whole-brain analysis is a promising strategy for gaining a deeper insight into events leading to addiction development and its particular aspects, such as relapse. We used a mouse model of alcohol addiction to perform whole-brain analysis of the activity during addiction‑like behaviors. Mice were trained to drink ethanol in “drinking in the dark” paradigm adapted to the IntelliCage system. Afterwards, the animals were deprived of alcohol and tested for relapse. Whole hemispheres of each mouse were subjected to optical clearing, c-Fos staining, light-sheet imaging, and analyzed for c-Fos expression to identify brain areas involved in relapse.