The dendritic tress of purkinje cells: A computer assisted analysis of HRP labeled neurons in organotypic cultures of kitten cerebellum

Abstract

Purkinje cell dendritic arborizations were studied in HRP intracellularly stained mature neurons grown during 30–50 days in organotypic cerebellar cultures from newborn kittens. The effects of afferent fiber depletions on the final dendritic topological parameters were investigated by computer assisted methods. Three differently deafferented models lacking both parallel and climbing fibers (PF and CF), only PF or only CF were thus studied. Qualitative data were found to be common to these 3 in vitro models and similar to those following other in vivo deafferenting procedures. Quantitative data obtained from 16 measurable parameters showed that all dendritic trees were markedly reduced in size as indicated by a decrease in total dendritic length and total number of segments although in all models individual segment lengths remained largely unaltered. A discriminant analysis permitted the recognition of 3 populations (100% well classified cells) based on 3 selected variables: dendritic field area, mean path length and total number of segments (with the lowest values for the model lacking only PF). These results are shown to lend further support to the role played in the final dendritis arrangement by both intrinsic (non-discriminant) and extrinsic (discriminant) factors and more precisely by the two distinct specific afferent systems (PF and CF).