Pharmacological characterization of ionotropic glutamate receptors in the zebrafish olfactory bulb

Abstract

The distribution of N-methyl- d-aspartate- (NMDA) and kainic acid- (KA) sensitive ionotropic glutamate receptors (iGluR) in the zebrafish olfactory bulb was assessed using an activity-dependent labeling method. Olfactory bulbs were incubated with an ion channel permeant probe, agmatine (AGB), and iGluR agonists in vitro, and the labeled neurons containing AGB were visualized immunocytochemically. Preparations exposed to 250 μM KA in the presence of a NMDA receptor antagonist ( d-2-amino-5-phosphono-valeric acid) and an α-amino-3-hydroxyl-5-methylisoxazole-4-propionic acid (AMPA) receptor antagonist (sym 2206), revealed KA receptor-mediated labeling of approximately 60–70% of mitral cells, juxtaglomerular cells, tyrosine hydroxylase-positive cells and granule cells. A higher proportion of ventral olfactory bulb neurons were KA-sensitive. Application of 333 μM NMDA in the presence of an AMPA/KA receptor antagonist (6-cyano-7-nitroquinoxaline-2,3-dione) resulted in NMDA receptor-mediated labeling of almost all neurons. The concentrations eliciting 50% of the maximal response (effective concentration: EC 50s) for NMDA-stimulated labeling of different cell types were not significantly different and ranged from 148 μM to 162 μM. These results suggest that while NMDA receptors with similar binding affinities are widely distributed in the neurons of the zebrafish olfactory bulb, KA receptors are heterogeneously expressed among these cells and may serve unique roles in different regions of the olfactory bulb.