Computer-assisted analysis of the developing Purkinje neuron. I. Effects of the age of the animal at the moment of explantation on the subsequent dendritic development in organotypic cultures

Abstract

Purkinje cell dendritic arborizations were studied in intracellularly horseradish peroxidase-stained mature neurons grown during 30–40 days in organotypic cerebellar cultures from kittens of various ages. The effects of the age of the animal at the moment of explantation upon the subsequent dendritic and axonal development were studied in kitten cerebella of 1, 12 and 21 postnatal days old. These effects were investigated by computer-assisted methods. Qualitative data were found to be different in these 3 in vitro models explanted at 3 different ages except for the spine development which remained the same. Quantitative data, obtained from 15 measurable parameters, showed that the degree of dendritic development was higher for the Purkinje cells explanted at the latest postnatal age: the older the animal, the larger the dendritic tree in culture. A discriminant analysis permitted a recognition of the 3 differently aged populations (94% well classified cells) based on 3 selected variables, number of primary dendritic trunks, number of rows and dendritic field areas. These results lend further support to the major role played in the final dendritic arrangement by extrinsic (discriminant) factors, their pre-existence in vivo being responsible for the subsequent degree of the in vitro development of the Purkinje cell dendritic arbor.