Distribution of corticotropin-releasing factor and calcitonin gene-related peptide in the developing mouse cerebellum.

Corticotropin-releasing factor (CRF)-like immunoreactive (IR) fibers were investigated ontogenically in the mouse cerebellum. CRF-IR was detected in the climbing fiber and mossy fibers as in other species. In addition, CRF-IR dense fiber plexuses were detected from postnatal day (PD) 2 to 9, in the developing Purkinje cell layer of the vermal lobules, paraflocculus, flocculus and crus 1 ansiform lobule, gradually forming a pericellular nest around the Purkinje cell somata. Immunoelectron-microscopical analysis showed that dense fibers made synaptic contacts with the Purkinje cell somata on PD 7. In the lobules mentioned above, CRF-IR dense fibers showed parasagittal banded patterns. Calcitonin gene-related peptide (CGRP)-IR showed similar fiber bands at these stages. Interestingly, these two patterns of peptidergic fiber bands were complementary in distribution. From around PD 9, CRF-IR fibers lost the immunoreactive dots in the Purkinje cell layer. Immunoreactivity at this stage was observed in the axons projecting to the molecular layer, and thin CRF-IR fibers began to appear in the neighboring area. Numerous typical climbing fiber-like CRF-IR fibers were found throughout the cerebellar cortex from PD 16 to adult. The inferior olivary complex (the origin of climbing fibers) appears to be the origin of these dense fiber plexuses as CRF-IR cells were already present from PD 2 in the dorsal cap nucleus, beta subnucleus and caudomedial part of the accessory olivary nucleus. No neurons containing both CRF and CGRP immunoreactivities were observed. These results suggest that CGRP- and CRF-IR developing climbing fibers innervate different compartments of Purkinje cells, especially in the vestibular cerebellar cortex in mice. Furthermore, CRF-IR fibers gradually changed to become typical climbing fibers, while CGRP-IR disappeared altogether.