Correlative microscopy of Purkinje dendritic spines: a field emission scanning and transmission electron microscopic study.

Abstract

Purkinje dendritic spines (Pds) of mouse cerebellar cortex were examined by field emission scanning electron microscopy (FESEM) and by transmission electron microscopy (TEM) using ultrathin sections and freeze-etching replicas, to study their three-dimensional features and intramembrane morphology. FESEM showed unattached mushroom-type, elongated and lanceolate Pds separated by 100-500 nm on the dendritic shaft surface. High resolution FESEM showed 25-50 nm globular subunits at the spine postsynaptic density corresponding to the localization of postsynaptic proteins and/or postsynaptic receptors. TEM images of ultrathin sections showed gem-like, mushroom-shaped, lanceolate and neckless or stubby spines. Freeze etching replicas exposed postsynaptic intramembrane particles that can be correlated with the globular subunits observed at high resolution FESEM. Parallel and climbing fiber endings were observed making asymmetric synaptic contacts with the Pds heads. Simultaneous contacts with the necks and heads were also found. The variety of Pds shapes were interpreted as spine conformational changes related with spine dynamic, and spine plasticity.