Glutamate-mediated release of Ca2+ in mitral cells of the olfactory bulb

1. Effects of glutamate on intracellular Ca2+ concentration ([Ca2+]i) were investigated in cultured olfactory bulb neurons of Xenopus laevis tadpoles. We imaged [Ca2+]i in these cells with the use of a confocal laser scanning microscope and the calcium indicator dyes Fluo3 and FuraRed. 2. In the standard bath solution, application of glutamate through a pipette resulted in an increase of [Ca2+]i in both mitral/ tufted (M/T) cells and interneurons. The increase occurred in all compartments of the cells, although in a nonhomogenous way. 3. In an ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid-buffered bath solution (Ca(2+)-free Ringer solution), glutamate reproducibly induced an increase of [Ca2+]i in M/T cells but not in interneurons. This increase in [Ca2+]i had the following properties: 1) it was delayed with respect to the combined ionotropic and metabotropic response to glutamate, 2) in some cases it stopped before the end of the glutamate application, and 3) it was not affected by D,L-2-amino-5-phosphonopentanoic acid and 6-cyano-7-nitroquinoxaline-2,3-dione added to the bath. It was interpreted as a release of Ca2+ from intracellular calcium stores. 4. Of 47 M/T cells that showed a glutamate-mediated release of [Ca2+]i, 46 also showed Ca2+ influx through ionotropic glutamate receptors, so that intracellular release of [Ca2+]i appeared to be associated with the presence of glutamate-gated ion channels. However, of 110 M/T cells showing an ionotropic response to glutamate, only 46 showed a glutamate-mediated release of [Ca2+]i. 5. The glutamate-mediated release of [Ca2+]i in the dendrites was higher than that in the soma. No indications for calcium waves were found. 6. Quisqualate and (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (ACPD) mimicked the effect of glutamate on the release of [Ca2+]i, with potencies quisqualate > glutamate > > ACPD. N-acetyl-aspartyl-glutamate did not induce release of [Ca2+]i. The release was partly blocked by (+)-alpha-methyl-4-carboxyphenylglycine. 7. In conclusion, some but not all M/T cells of the olfactory bulb of X. laevis show a glutamate-mediated and predominantly dendritic increase of [Ca2+]i that is associated with glutamategated channels. The pharmacological properties of the corresponding metabotropic glutamate receptor (mGluR) resemble those of the mGluR1/5 receptors in rat.