Figure 4: Perforant path fibers are present in the outer molecular layer (OML) during the development of RV-GFP-labeled GCs in vitro. | Scientific Reports

Figure 4: Perforant path fibers are present in the outer molecular layer (OML) during the development of RV-GFP-labeled GCs in vitro.

From: Time-lapse imaging reveals highly dynamic structural maturation of postnatally born dentate granule cells in organotypic entorhino-hippocampal slice cultures

Figure 4

(a) The biotinylated dextran amine tracer Mini Ruby was applied to the entorhinal cortices (ECs) of OTCs at DIV 15 and images were taken on DIV 17 showing the axons of EC neurons projecting to the molecular layer (ML) of the dentate gyrus (red). In the granule cell layer (GCL), mature neurons are labeled with the marker calbindin (CB), while immature neurons are labeled with doublecortin (DCX). (b) Z-projection of two images showing an 8-day old OTC treated with the RV (green) and Mini Ruby (red). Mini Ruby was applied on DIV 3 and OTCs were fixed on DIV 8. Prox1-positive (blue) 8-day-old postnatally born RV-GFP-labeled GCs directed their growing dendrites toward the OML (white arrowheads) that was already re-innervated by Mini Ruby-labeled entorhinal fibers. (c) Z-projection of four images (z-axis interval: 1 μm) of an RV and Mini Ruby treated OTC at DIV 20. A 20-days-old RV-GFP-labeled GC exhibiting an elaborate dendritic arbor that extended well into the OML (white arrows) where labeled perforant path axons were present. Scale bars: (a) 50 μm, (b,c) 10 μm.

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