The innervation of calcitonin gene-related peptide to the Purkinje cells and granule cells in the developing mouse cerebellum

Abstract

The present study analyzed the ontogeny of calcitonin gene-related peptide-like immunoreactive (CGRP-IR) structures in the mouse cerebellum. No CGRP-IR neurons were detected at any stage, but three types of CGRP-IR fibers were seen: (1) CGRP-IR dense fiber plexuses which appeared transiently in the developing cerebellum, (2) thin varicose fibers, and (3) mossy fiber-like fibers. The CGRP-IR dense fiber plexuses appeared in the developing Purkinje cell layer at postnatal day 2. From postnatal days 6 to 11, these fibers formed pericellular nests around Purkinje cells. After that stage, these fibers rapidly disappeared and no such plexuses were seen in the adult cerebellum. CGRP-IR fiber plexuses were not evenly distributed, and they had a parasagittal banded pattern in the frontal sections. These plexuses existed in the region of all vermis, crus 1 of the ansiform lobe, simplex lobule, and flocculus, while the other lobules were devoid of such fibers. Under electron microscopy, these CGRP-IR fibers were seen to make synaptic contacts with somatic spines of Purkinje cells, suggesting that CGRP-IR plexuses were closely related to the developing Purkinje cells. Mossy fiber-like CGRP-IR fibers appeared in the granular layer on postnatal day 2, and increased in number to reach a peak on postnatal day 12. Thereafter, they decreased slightly to reach a plateau on postnatal day 30. Under electron microscopy these CGRP-IR fibers were revealed to be the mossy fibers which regulate the granule cells. Thin varicose CGRP-IR fibers were rarely seen at birth, but on postnatal day 8, many fibers appeared in all layers and increased by postnatal day 30. They distributed equally throughout the cerebellar cortex with a slight predominance in density in the molecular and Purkinje cell layer. Immunoelectron microscopic analysis showed that these fibers made synaptic contacts with small dendrites in the molecular layer.