Synaptic and neurochemical features of calcitonin gene-related peptide containing neurons in the rat accessory optic nuclei

Abstract

Within the rodent visual system, calcitonin gene-related peptide (CGRP) is selectively expressed in neurons in the accessory optic nuclei (AON), including the dorsal terminal nucleus (DTN), lateral terminal nucleus (LTN) and medial terminal nucleus (MTN). To determine whether CGRP-immunoreactive neurons are involved in visual circuitry, electron microscopic preparations were analyzed from normal rats and rats with optic nerve transections. A co-localization analysis was also made because CGRP-labeled neurons had features of GABAergic neurons. Thus, sections were prepared for light microscopy to determine whether CGRP-containing neurons also had glutamate decarboxylase (GAD) and other markers for GABAergic neurons, such as calcium binding proteins: calbindin (CB), calretinin (CR) and parvalbumin (PV). Electron microscopy of the DTN and LTN showed CGRP-labeled somata and dendrites that were postsynaptic to axon terminals forming asymmetric synapses. Many of these axon terminals degenerated following optic nerve transection indicating that retinal ganglion cells form synapses with CGRP-labeled neurons in the AON. In the DTN, LTN and MTN, CGRP-labeled axon terminals formed symmetric synapses with unlabeled somata as well as dendritic shafts and spines. Consistent with this type of synapse being GABAergic were the co-localization data showing that about 90% of the CGRP-labeled neurons co-localized GAD in the AON. Many CGRP-labeled neurons showed immunostaining for CR (40%) whereas only a few had labeling for CB (5%). No CGRP-labeled neurons had PV. These data show that CGRP-containing neurons receive direct retinal input and represent a subpopulation of GABAergic neurons which differentially co-express calcium-binding proteins.