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Chemical Senses Advance Access originally published online on February 25, 2008
Chemical Senses 2008 33(4):397-404; doi:10.1093/chemse/bjn007
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© The Author 2008. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Electroolfactogram Responses from Organotypic Cultures of the Olfactory Epithelium from Postnatal Mice

Giulietta Pinato, Juraj Rievaj, Simone Pifferi, Michele Dibattista, Lara Masten and Anna Menini

Neurobiology Sector, International School for Advanced Studies, SISSA, Italian Institute of Technology, SS 14 Km 163.5, 34012 Basovizza, Trieste, Italy

Correspondence to be sent to: Anna Menini, Neurobiology Sector, International School for Advanced Studies, SISSA, SS 14 Km 163.5, 34012 Basovizza, Trieste, Italy. e-mail: menini{at}sissa.it


  Abstract

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 µM 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 µM 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.

Key words: electroolfactogram, IBMX, olfaction, organotypic culture, transduction

Accepted 17 January 2008


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