ISOLATION OF MATURE OLFACTORY NEURONS USING RETROGRADE LABELING AND FLOW-CYTOMETRY

Currently we can observe an increasing interest for epithelial olfacto ry neurones. Several laboratories are involved in elucidation of the o dorant-molecule recognition process and transduction cascade which bri ngs information to the olfactory bulb. Others use this model, unique i n mammals, to accumulate new knowledge on the neurogenesis phenomenon. Here we describe a simple and efficient method to purify mature olfac tory neurone populations extracted from the nasal cavity. The approach relies on retrograde axonal tracing followed by flow cytometry sortin g. For this purpose we inject a fluorescent dye (Fast Blue or diI C-18 (3)) in the olfactory bulb of adult rats. Seven days later, we extract the nasal turbinates and separate the mucosa from the subjacent lamin a propria. The tissue is enzymatically dissociated and the labelled ce lls are sorted with a flow cytometer. Purification of the mature olfac tory neurones varies from 80 to 99%.