Intracellular injections of lucifer yellow into lightly fixed mitral cells reveal neuronal dye-coupling in the...

We report that intracellular injections of Lucifer Yellow into lightly fixed mitral cells revealed dye-coupling between mitral cells and between mitral and granule cells in the form of discrete, radially oriented cell clusters. Dye-coupling was observed in animals as early as postnatal day 10 (P10) and at least until P30. In P10 rats, a mean of 2.5 dye-coupled mitral cells and 27 granule cells were observed per column. Mean column depth and width were 169 microns and 86 microns respectively. Most of the dye-filled granule cells were found within 150 microns of the mitral cell layer. No significant changes were found at P20. By P30, the mean number of granule cells per column increased to 42 and the addition of granule cells occurred in areas proximal to the mitral cell layer. Immunocytochemical results indicate that the developing bulb contains a large concentration of the gap junction protein Connexin 43 (Cx43). Cx43-like immunoreactivity was found at all ages examined, with the most intense staining in the nerve and glomerular layers. Less intense Cx43-like immunoreactivity was found in both the mitral and granule cell layers, with Cx43-like immunoreactive puncta observed between and around cell body profiles. Freeze-fracture analysis revealed the presence of gap junction-like plaques on mitral cells, further suggesting that the dye-coupling occurred across interneuronal gap junctions. Neuronal coupling during development could provide an inter-cellular pathway for the passage of relevant developmental signals which could influence the formation and/or strengthening of synaptic contacts. The coupling could also be involved in the synchronization of neuronal activity, which may be important for olfactory coding.