Unipolar brush cells develop a set of characteristic features in primary cerebellar cultures

The unipolar brush cells (UBCs) are a class of excitatory interneurons recently discovered in the cerebellar granular layer. UBCs differ morphologically and biochemically from granule cells, although they share the same mossy fiber and Golgi cell inputs. To elucidate development of the UBCs, we sought to ascertain their presence in primary cerebellar cultures and the class-specific properties they develop in vitro outside of the context of the tissue. By light and electron microscopy, we demonstrate that primary cultures from embryonic and postnatal mouse and rat cerebella contain UBC-like neurons that are highly polarized and can be distinguished from granule cells on several grounds. Granule cells are more numerous in dissociated postnatal cultures than in embryonic cultures; express little, if any, calretinin immunoreactivity; and develop dendritic processes devoid of typical claw-like endings, but provided with small synaptic junctions. By contrast, UBC-like neurons occur more frequently in embryonic cultures than in postnatal cultures, are intensely calretinin-positive, and develop characteristic cell organelles, dendrites, and large synapses. In embryonic cultures, the UBC-like neurons have a clear nucleus and contain a special cytoplasmic array of ringlet subunits, resembling the botrysome. At 12-28 days in vitro, the UBC dendrites contain abundant mitochondria, are provided with clusters of non-synaptic appendages, and engage in glomerular arrays together with large and small axon terminals. The large terminals contain round synaptic vesicles, form extensive, asymmetric synapses with the cell bodies and the dendrites of the UBC-like neurons, and resemble mossy terminals, while the small terminals contain pleomorphic vesicles, form symmetric synaptic junctions, and resemble Golgi terminals. In postnatal cultures grown for 12 days, UBC-like neurons are rare and resemble in most aspects the cells observed in embryonic cultures, although they rarely develop elaborate dendritic brushes.