Ontogenesis of enkephalinergic afferent systems in the opossum cerebellum

Enkephalin (ENK) immunoreactive climbing fibers, mossy fibers and a beaded plexus of axons are present in the adult opossum's cerebellar cortex. We have used the indirect antibody peroxidase-antiperoxidase technique to study the ontogeny of enkephalinergic axons in the cerebellum of pouch young opossums from postnatal day (PD) 1 to PD 83. On PD 1, ENK axons are present in the intermediate layer of the cerebellar anlage. At PD 18, after a period of 'waiting', ENK fibers form clusters throughout the cerebellar cortex primarily within the nascent Purkinje cell layer. By PD 40, axon terminals with a climbing fiber phenotype circumscribe Purkinje cells; immature mossy fiber rosettes are present within the internal granule cell layer. A third axon phenotype, beaded ENK fibers can be distinguished on PD 68. Between PD 40 and PD 68, the distributions of ENK climbing and mossy fibers overlap in vermal lobules II-VIII and X, whereas in the hemispheres climbing fibers predominate. However, by PD 83, ENK positive climbing fibers are no longer evident in lateral folia. These results indicate that early arriving ENK axons are present before the differentiation of their cellular targets. Further, a transient appearance of ENK in discrete populations of developing climbing fibers suggests several developmental events: (1) cell death in the inferior olive, (2) collateral regression, or (3) a transient expression of this peptide, that may be characteristic of this chemically defined system of axons.