Lesion‐induced mossy fibers to the molecular layer of the rat fascia dentata: Identification of postsynaptic granule cells by the Golgi‐EM technique

The axons of the dentate granule cells, the hippocampal mossy fibers, sprout “backward” into the dentate molecular layer when this is heavily de‐nervated. Using the combined Golgi‐electron microscopy (EM) technique we now demonstrate that these aberrant supragranular mossy fibers at least in part terminate on granule cell dendrites. Sprouting of mossy fibers into the dentate molecular layer was induced in adult rats by simultaneous surgical removal of the commissural and ento‐rhinal afferents to the fascia dentata. After at least 7 weeks survival, the presence of mossy fiber terminals in the inner part of the dentate molecular layer was demonstrated by light microscopy of Timm‐stained cryostat sections and by electron microscopy. In the electron microscope the mossy fiber terminals were identified by their unique structural characteristics, namely, the unusually large size of the terminals, the dense packing of clear synaptic vesicles with a few dense core vesicles intermingled, the presence of asymmetric synaptic contacts with spines and desmosome‐like contacts with dendritic shafts, and the continuity with a thin unmyelinated preterminal axon. Golgi‐stained granule cells were first identified in the light microscope, and then, after deimpregnation, the same cells were examined in the electron microscope. In ultrathin, serial sections lesion‐induced mossy fiber terminals were found in synaptic contact with spines on proximal dendritic segments of such identified Golgi‐impregnated granule cells. From this we conclude that the aberrant, supragranular mossy fibers can innervate dendrites of the parent cell group, the dentate granule cells. The results, moreover, provide an example of reactive synaptogenesis where both the sprouted afferents and its postsynaptic element have been identified.