Development of the mossy fibers of the dentate gyrus: I. A light and electron microscopic study of the mossy fibers and their expansions.

The postnatal development of the axons of the dentate granule cells--the so-called mossy fibers--was studied at the light microscopic level in Timm and Golgi preparations and also by transmission electron microscopy. In the Timm-stained material, there was a distinctive coloration in the hilus and incipient stratum lucidum, indicating the presence of mossy fibers, on the first postnatal day. Over the next two weeks, the stained areas became more extensive, the size and density of the stained particles increased, and the particles became more intensely stained. These signs of progressive development of the mossy fibers appeared to reflect, temporally and topographically, the developmental gradients followed by their parent granule cells. The Golgi material confirmed the presence of mossy fibers in the hilus on the first postnatal day. Fasciculi of mossy fibers were observed in the stratum lucidum of the 3-day-old hippocampus, and although these immature axons were devoid of large synaptic expansions, they did have prominent growth cones at their termini. Small expansions along the lengths of the axons first appeared on day 7 and these grew to approximately an adult size and complexity by about day 14. The postsynaptic component of the mossy fiber synapse, the "thorny excrescence," did not begin to emerge from the proximal portion of the pyramidal cell dendrites until sometime after day 9. At the electron microscopic level we observed, on the first postnatal day, small, immature mossy fiber expansions which made both symmetric and asymmetric contacts directly with dendritic shafts. These profiles, which were only one tenth the size of mature expansions, grew rapidly between postnatal days 1 and 9 and increased their mean area by a factor of five. On or about day 9, as the "thorny excrescences" emerged, the asymmetric synapses came to be associated with these spinous processes. Taken together, the Golgi and electron microscopic analyses support the suggestion that mossy fibers establish synaptic contact with pyramidal cell dendrites early in the postnatal period, several days before there is any indication of spine development. Furthermore, the "thorny excrescences" develop after the more typical, pedicellate spines have appeared on the distal pyramidal cell dendrites. Finally, while it is clear that the mossy fibers in our 21-day-old material are, for the most part, fully matured, a more subtle and protracted development of the system, long into adulthood, is indicated by the increased area and density of stained particles in the Timm preparations from adult animals.