Early dendritic development of Purkinje cells in the rat cerebellum. A light and electron microscopic study using axonal tracing in 'in vitro' slices

Abstract

The early stages in the formation of Purkinje cell dendritic arbors have been analyzed using the horseradish peroxidase (HRP) 'in vitro' axonal tracing method, from embryonic day 19 (E19) to postnatal day 6 (P6). These stages comprise the transition from the bipolar Purkinje cell, at the end of its migration, to the phase of stellate cell with disoriented dendrites. Postmigratory Purkinje cells in the cortical plate exhibit poorly elaborated bipolar shapes, here named 'simple-fusiform' cells. They constitute the vast majority of labeled cells up to P0, and thereafter they decrease in number until P4. As a result of continuous outgrowth of new primary dendrites emerging from the apical pole but also from the basal and lateral aspects of the cell bodies, the Purkinje cells enter the 'complex-fusiform' phase, which peaks by P1 and slowly disappears by P6. The disappearance of 'complex-fusiform' cells is the result of an intense regressive process with resorption or retraction of the long dendrites that reaches a maximum by P3. We have called this stage: the Purkinje cell with 'regressive-atrophic' dendrites. This regression marks the initiation of the phase of the stellate cell, characterized by the explosive outgrowth of shorter perisomatic protrusions emerging in all directions. By P6, almost all the labeled Purkinje cells have attained this phase. The ultrastructural study of the labeled Purkinje cells has revealed that the transient dendrites of the fusiform cells have all the cytologic features of mature dendrites, particularly cytoskeletal elements (microtubules) and free polyribosomes. More importantly, axon terminals of unknown origin establish a few, constantly present, mature-like synaptic contacts on the dendritic shafts and spinous protrusions from P0, the earliest studied age. Their frequency increases on the Purkinje cells which enter the phase of stellate cell. Our results emphasize that the transformation of bipolar postmigratory Purkinje cells into the stellate cell stage results from a complex cascade of alternating creative and destructive processes, taking place in parallel with the formation and regression of mature synaptic contacts, between the remodelling dendritic arbors and unidentified afferent inputs. Purkinje cells, in all the different transitional stages, are present side by side in the same folial regions, at least until P4, and receive a similar contingent of synaptic input. This indicates that the dendritic remodelling is not driven by the synaptic inputs, but obeys either neural interactions that lead Purkinje cells to assume their monocellular layer configuration, or an internal clock depending on the Purkinje cell birthdate, or an interplay between these two kinds of mechanisms.