From hours to days: prolonging the functional viability of acute brain slices to >36 h

Abstract

We have developed an experimental protocol for acute brain slices of juvenile and adult mice which, in our hands, enables whole-cell patch-clamp experiments on healthy neurons with intact electrophysiological characteristics for more than 36 hours after preparation. This drastically expands the conventional viability window of brain slices of 8-12 hours, using only standard laboratory equipment and procedures. We successfully applied this protocol to evaluate neurons of the medial prefrontal cortex (mPFC), motor cortex (M1/M2), globus pallidus (GP), and thalamic reticular nucleus (TRN) of mice aged P11 to ~2 years postnatal, demonstrating the robustness of our method. By prolonging the usability of the preparation, this procedure a) reduces the number of animals used per study, b) yields healthy brain slices also from adult animals, c) allows prolonged experimental procedures, and d) reduces material cost and preparation effort for the experimenter.