Postnatal maturation of rat Purkinje cells cultivated in the absence of two afferent systems: An ultrastructural study

Abstract

Organized cultures of newborn rat cerebellum were established in Maximow chambers in order to study the maturation of Purkinje cells in absence of afferent systems. In the first model, standard cultures were devoid of extracerebellar afferents mossy and climbing fibers. Despite this absence, somatic spines appeared upon Purkinje cells during the first week in vitro and maturation proceeded normally except for the almost absence of spiny branchlets. Large dendritic trunks were studded with numerous spines, some of which were naked, a few bearing isolated post-synaptic densities and others occupied by boutons of parallel fibers. Stellate and basket axons made synapses upon the smooth portions of dendrites and soma. In a second model, the cultures were fed the antimitotic drug methylazoxymethanol (MAM) to prevent multiplication of granule cell precursors. Despite the absence of climbing and parallel fibers, the elongation of Purkinje dendrites was not prevented, but again the dendritic arbor consisted of large trunks studded with spines; somatic as well as dendritic spines were contacted by large boutons identified as Purkinje recurrent collaterals (PRC). It is concluded that the Purkinje cell possesses a large autonomy from afferent systems as to the growth of soma and dendrites. Conversely, the geometry of the dendrite and especially the spiny branchlets depend on the presence of both climbing and parallel fibers. One may conclude from the above experiments that specificity of synaptic contacts is maintained as long as postsynaptic sites are not devoid of their normal afferents. Heterologous synapses are formed when postsynaptic sites are present, their normal afferents absent and aberrant ones increasing by collateral sprouting. Such is probably the case in the second model of this study.