Odorant/receptor binding and initial olfactory information processing occurs in olfactory receptor neurons (ORNs) within the olfactory epithelium. Subsequent information coding involves high-frequency spike synchronization of paired mitral/tufted cell dendrites within olfactory bulb (OB) glomeruli via positive feedback between glutamate receptors and closely-associated gap junctions. With mRNA for connexins Cx36, Cx43 and Cx45 detected within ORN somata and Cx36 and Cx43 proteins reported in ORN somata and axons, abundant gap junctions were proposed to couple ORNs. We used freeze-fracture replica immunogold labeling (FRIL) and confocal immunofluorescence microscopy to examine Cx36, Cx43 and Cx45 protein in gap junctions in olfactory mucosa, olfactory nerve and OB in adult rats and mice and early postnatal rats. In olfactory mucosa, Cx43 was detected in gap junctions between virtually all intrinsic cell types except ORNs and basal cells; whereas Cx45 was restricted to gap junctions in sustentacular cells. ORN axons contained neither gap junctions nor any of the three connexins. In OB, Cx43 was detected in homologous gap junctions between almost all cell types except neurons and oligodendrocytes. Cx36 and, less abundantly, Cx45 were present in neuronal gap junctions, primarily at "mixed" glutamatergic/electrical synapses between presumptive mitral/tufted cell dendrites. Genomic analysis revealed multiple miRNA (micro interfering RNA) binding sequences in 3′-untranslated regions of Cx36, Cx43 and Cx45 genes, consistent with cell-type-specific post-transcriptional regulation of connexin synthesis. Our data confirm absence of gap junctions between ORNs, and support Cx36- and Cx45-containing gap junctions at glutamatergic mixed synapses between mitral/tufted cells as contributing to higher-order information coding within OB glomeruli.
|Original language||English (US)|
|Number of pages||35|
|Journal||Journal of Neurocytology|
|State||Published - Sep 2005|
Bibliographical noteFunding Information:
This work was supported by NIH grants NS31027, NS44010, and NS 44395 (JER), DC00566, DC04657(DR) and DC04952 (CZ) and by CIHR (JIN). We thank B. McLean for excellent technical assistance, Dr. D. Paul (Harvard University) for providing us with breeding pairs of wild-type and Cx36 knockout mice, Brian Gebhardt and Jennifer A. Sampson for assistance in preparing stereoscopic images, Elek Molnar (University of Bristol) for providing antibodies to AMPA receptors, and Elliot Hertzberg (Albert Einstein School of Medicine) for providing anti-connexin Cx43 antibody Ab18A. We also thank Dr. Torgeir Holen (University of Oslo) for useful comments regarding miRNA.