Electroolfactogram responses from organotypic cultures of the olfactory epithelium from postnatal mice.
Organotypic cultures of the mouse olfactory epithelium connected to the olfactory bulb were obtained with the roller tube technique from postnatal mice aged between 13 and 66 days. To test the functionality of the cultures, we measured electroolfactograms (EOGs) at different days in vitro (DIV), up to 7 DIV, and we compared them with EOGs from identical acute preparations (0 DIV). Average amplitudes of EOG responses to 2 mixtures of various odorants at concentrations of 1 mM or 100 microM decreased in cultures between 2 and 5 DIV compared with 0 DIV. The percentage of responsive cultures was 57%. We also used the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) to trigger the olfactory transduction cascade bypassing odorant receptor activation. Average amplitudes of EOG responses to 500 microM IBMX were not significantly different in cultures up to 6 DIV or 0 DIV, and the average percentage of responsive cultures between 2 and 5 DIV was 72%. The dose-response curve to IBMX measured in cultures up to 7 DIV was similar to that at 0 DIV. Moreover, the percentage of EOG response to IBMX blocked by niflumic acid, a blocker of Ca-activated Cl channels, was not significantly different in cultured or acute preparations.
Pinato
Giulietta
G
Neurobiology Sector, International School for Advanced Studies, SISSA, SS 14 Km 163.5, 34012 Basovizza, Trieste, Italy.
Rievaj
Juraj
J
Pifferi
Simone
S
Dibattista
Michele
M
Masten
Lara
L
Menini
Anna
A
D000818:Animals; D000831:Animals, Newborn; D018118:Chloride Channels / Q000037:antagonists & inhibitors; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D051379:Mice; D008807:Mice, Inbred BALB C; D009812:Odorants / Q000032:analysis; D009831:Olfactory Mucosa / Q000201:enzymology / Q000378:metabolism / Q000502:physiology*; D009924:Organ Culture Techniques; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D012903:Smell / Q000502:physiology*; D013997:Time Factors
D018118:Chloride Channels; D010726:Phosphodiesterase Inhibitors; D010727:Phosphoric Diester Hydrolases