Fig. 1

Entorhinal denervation in vitro model. a Schematic of an organotypic entorhino-hippocampal slice culture. The entorhino-hippocampal projection (red), which originates in the entorhinal cortex (EC) and terminates in the outer molecular layer (OML) of the dentate gyrus (DG) is transected with a sterile scalpel (black line; plane of transection, top). This lesion leads to a partial denervation of dentate granule cells (green schematic cell shown in the magnification of the DG, bottom) without directly damaging the target region (CA1, hippocampal subfield Cornu Ammonis 1; CA3, hippocampal subfield CA3; GCL, granule cell layer; IML, inner molecular layer; OML, outer molecular layer). b A non-denervated (top) and denervated (bottom) three-week old slice culture stained with TO-PRO (blue, nuclear stain). To assure a complete and reproducible denervation of the DG in all experiments, the EC was removed from the culturing dish. The inset shows Mini-Rubi traced (red) entorhinal fibers terminating in the OML of the DG. Scale bar: 200 μm (inset: 50 μm). c Entorhino-hippocampal slice cultures prepared from Thy1-GFP mice were employed to visualise individual dentate granule cells of denervated cultures and age- and time-matched non-denervated controls using time-lapse microscopy. An example of a GFP-expressing granule cell is shown (2D-projected confocal image stack). Dendritic trees of dentate granule cells were manually reconstructed in 3D-confocal image stacks. Scale bars: 100 μm