Protocol for whole-cell patch-clamp recording and post hoc identification of hippocampal CA2 pyramidal neurons in adult mouse brain slices

We present a protocol for whole-cell recordings from hippocampal CA2 pyramidal neurons in acute adult mouse brain slices. We describe the preparation of viable dorsal hippocampal slices, the visual identification and recording of CA2 neurons, and intracellular labeling. We then detail post hoc immunohistochemical verification using CA2-specific markers. This approach enables reliable functional characterization of CA2 neurons and unambiguous confirmation of their anatomical localization. For complete details on the use and execution of this protocol, please refer to Le Bail et al. 1 • Preparation of acute dorsal hippocampal brain slices from adult mouse brain • Whole-cell patch-clamp recording from visually identified CA2 pyramidal neurons • Post hoc molecular identification and localization of recorded CA2 pyramidal neurons Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. We present a protocol for whole-cell recordings from hippocampal CA2 pyramidal neurons in acute adult mouse brain slices. We describe the preparation of viable dorsal hippocampal slices, the visual identification and recording of CA2 neurons, and intracellular labeling. We then detail post hoc immunohistochemical verification using CA2-specific markers. This approach enables reliable functional characterization of CA2 neurons and unambiguous confirmation of their anatomical localization.