The organization and development of the hippocampal mossy fiber system.

The anatomical organization and development of the hippocampal mossy fiber system has been reviewed with special reference to its organization in the common laboratory rat. The mossy fibers originate from the granule cells of the dentate granular layer and the few granule cells found scattered in the dentate molecular layer and hilus. Via a complex system of collaterals the mossy fibers terminate on several types of neurons in the hilus, e.g. the basket cells and the mossy cells. Upon leaving the hilus to pass into Ammon's horn, the mossy fibers converge to form a distinct band of fibers that terminates on the proximal part of the apical and basal dendrites of the pyramidal and basket cells of the regio inferior. In some mammalian species the mossy fibers may continue into the adjacent part of the regio superior. Despite differences in the number of granule cells and pyramidal cells at different septotemporal levels this organization is relatively uniform along the septotemporal extent of the hippocampus. During development the mossy fibers grow out in a sequential manner that matches the pattern of neurogenesis and the aggregation of the cells of origin. From the level at which they originate, the fibers diverge along the septotemporal axis in such a way that the oldest granule cells have the most extensive projections. The adult topographic organization, which is already apparent at the earliest developmental stages, is thus formed in a stepwise fashion. It is concluded that the organization of the hippocampal mossy fibers indicates that neuronal specificity should not be explained by cellular recognition alone, but rather as the cumulated product of the preceding sequence of developmental events that include neurogenesis, migration, aggregation and directed axonal outgrowth.

Neonatal hippocampal neurons, retrogradely labeled with granular blue, survive intracerebral grafting and explantation to tissue culture.

Previous studies have shown that developing neocortical neurons labeled in situ by retrograde axonal transport of the fluorescent dye Granular Blue can retain this dye for at least 2 months essentially without fading or leakage. In this study, retrograde labeling with Granular Blue was used to label neonatal rat hippocampal neurons prior to intracerebral grafting to uninjected littermates, and explantation as slice cultures. Hippocampal regio inferior and hilar neurons labeled through their developing commissural axons were found to survive axotomy and subsequent grafting and explantation for at least 3 and 8 weeks, respectively. The labeling helped define the developmental differentiation of the neurons at the time of manipulation and provides a new method for identification of a specific population of transplanted or explanted neurons.

The development of the dentate area and the hippocampal mossy fiber projection of the rat.

The development of the dentate area and the hippocampal mossy fiber system of the rat has been investigated at the light microscopic level by using fluorescent tracing, Nissl, and Timm's histochemical methods. Although the cytoarchitectonic development of the dentate granular layer is mainly a postnatal phenomenon, the initial events take place before birth. The aggregation and maturation of the cells in the granular layer proceed in a graded fashion from the lateral to the medial and from the superficial to the deep aspects of the layer. The earliest-formed granule cells are probably derived directly from the cells of the ventricular zone. They start to form mossy fibers prenatally, either during the relatively long period of migration to the granular layer or soon after their arrival. However, most of the granule cells are derived from a secondary proliferative center in the hilus. They start to produce mossy fibers postnatally a while after arriving at the granular layer. The total complement of granule cells starts to grow mossy fibers in a sequence that is related to the final position of the cells of origin within the granular layer. This sequence also proceeds in a graded fashion from the lateral to the medial and from the superficial to the deep aspects of the layer. In the beginning the mossy fibers elongate relatively rapidly. Already at birth the Timm-stained mossy fiber zone occupies the anterolateral part of the hilus and the adjacent suprapyramidal parts of the regio inferior. Once the mossy fibers have reached the distal end of the regio inferior they elongate along the longitudinal axis of the hippocampus more slowly. At the same time the Timm-stainability of the mossy fiber zone, which, during the first postnatal week, is weaker toward the regio superior, develops a mature pattern in which the distal part of the zone stains most intensely. Throughout development, fibers from the granule cells that form first are longer and diverge more in the septotemporal dimension than fibers from later-forming granule cells. In contrast to other axonal systems which appear to be sculptured from a diffuse set of connections the results presented here provide evidence that the topographic relationships of the mossy fiber system develop in a stepwise fashion.

The Timm-stained hippocampus of the European hedgehog: a basal mammalian form.

A quantitative and qualitative description has been made of the components of the Timm-stained hippocampus of the European hedgehog. While the laminar organization and the relative size of the major subdivisions of the hippocampus (i.e., area dentata, Ammon's horn, and subiculum) are similar to those of the albino laboratory rat, the relative proportions and the staining characteristics of some of the components of the subdivisions are different. The differences are particularly evident in Ammon's horn where regions are poorly differentiated along the dentatosubicular axis and the mossy fiber zone is relatively extensive. The description characterizes a hippocampal form that can be used as a basal reference in comparative studies of the mammalian hippocampus.

Ectopic granule cells of hilus fasciae dentatae projecting to the ipsilateral regio inferior of the rat hippocampus.

Retrograde fluorescent tracing techniques were used in a search of projections from the hilus of fascia dentata to the Ammon's horn. We found that up to 5% of the neurons in the hilus project to the ipsilateral regio inferior. The projection originates from cells that morphologically resemble ectopic granule cells and terminates in the mossy fiber layer. We found no evidence for a projection from the hilus to the ipsilateral regio superior or the contralateral Ammon's horn.

Hippocampal mossy fibers in the regio superior of the European hedgehog.

The hippocampal mossy fiber zone of the European hedgehog has been studied at the light and electron microscopic levels. In contrast to the organization of this zone in other mammals, the mossy fiber zone of the hedgehog is not confined to the region of Ammon's horn that is characterized by large pyramidal cells (i.e. regio inferior) but extends into the region that contains small pyramidal cells (i.e. regio superior). The terminals, throughout the relatively extensive mossy fiber zone of the hedgehog, have the large multivesicular, multicontact morphology that characterizes the mossy fiber terminals observed in other species and can be demonstrated to be the axon terminals of granule cells located in the fascia dentata. In view of the primitive organization of the forebrain of the European hedgehog, which is believed to be representative of that of the first mammals, the boundaries of the mossy fiber zone of these animals suggest that the mossy fibers extended over a larger part of Ammon's horn during the first phases of mammalian evolution and became restricted to the regio inferior during the early stages of mammalian evolution.

The hippocampal mossy fiber system of the rat studied with retrograde tracing techniques. Correlation between topographic organization and neurogenetic gradients.

The topographical organization of the hippocampal mossy fiber system, which connects the dentate granule cells with the pyramidal cells of the regio inferior, has been examined in rats with retrograde tracing methods. Following the application of the fluorescent dye True Blue to different parts of the mossy fiber layer in the hippocampal regio inferior, retrogradely labeled granule cells were observed in the dentate fascia. The distribution of the labeled cells within the dentate granule cell layer indicates that all mossy fibers have an almost parallel, slightly descending course in regio inferior near the dentate hilus. In the ventricular part of region inferior, particularly toward the transition to the regio superior, the mossy fibers are sorted out according to the position of their parent cell bodies within the granular layer. Near the transition to regio superior the fibers from lateral granule cells extend both septally and temporally over a longer distance than the fibers from more medial cells. Similarly, the fibers coming from the superficial cells extend both septally and temporally over a longer distance than those from the deep cells. The mossy fibers arising from a specific septotemporal level of the dentate fascia innervate a segment of the regio inferior that extends approximately 180 micrometer above to approximately 1,600 micrometer below the level of origin. Similar results were obtained following injections of Nuclear Yellow and horseradish peroxidase. Since previous studies have demonstrated that the granule cells are formed along gradients from lateral to medial and from superficial to deep, there appears to be a correlation between the formation, and hence the position, of the granule cells and the topography of their projection into the regio inferior.

Organization of the mossy fiber system of the rat studied in extended hippocampi. II. Experimental analysis of fiber distribution with silver impregnation methods.

The mossy fibers, a major intrinsic hippocampal pathway connecting the dentate granule cells with the pyramidal cells in CA4 and CA3, have been reexamined in rats using mainly Fink-Heimer silver impregnation methods for demonstration of degenerating axons. By extending isolated hippocampi and cutting sections normal to the long axis, simple two-dimensional reconstructions of both the lesions and the resultant degeneration could be made. In the hilus, the zone with the greatest concentration of degenerating boutons was found between the lesioned granule cells and the CA3 pyramidal cells abuting on the hilus; outside this zone the concentration declines rather rapidly. Degenerating boutons were also observed in low concentration up to 200-300 microgram septal and temporal to the lesion. The mossy fibers in CA3 nearest the hilus have an intrapyramidal course and display a lamellar organization with fibers from the granule cells of the medial blade lying deep to those from the dentate crest. These in turn lie deep to those from the graule cells of the lateral blade. A mediolateral difference in the projection of the graule cells on the CA3 pyramidal cells was discovered: fibers from the medial granule cells descend about 600 micrometer in the temporal direction, whereas fibers from the lateral granule cells descend about 1,200 micrometer. This causes a divergence of the fibers from one single level, especially of the part of the fibers, being farthest away from the hilus. The degree of descent of the fibers from each mediolateral position of the granule cells was constant at all septotemporal levels examined.