A preparation enriched in olfactory receptor cells has been obtained from the olfactory mucosa of the catfish (Ictalurus punctatus). The tissue was treated successively with trypsin, DNase, trypsin inhibitor, EDTA in Ca+ + , Mg+ + free medium according to a method derived from that of Cohen, et al.(1). After mechanical disruption of the isolated olfactory lamellae, the cells were isolated by centrifugation on a Ficoll gradient. Each type of cell was morphologically identified by comparing in situ and in vitro preparations by SEM. Small round cells were collected on 10% Ficoll. The nature of these cells is not known but part of them are certainly basal cells which have been shown(2) to be the precursors of the constantly regenerating olfactory neurons. Respiratory cells settled mainly on 20% Ficoll. A fraction containing 60% sustentacular cells was collected on 33% Ficoll. Olfactory cells characterized by an axon, a dendrite and several cilia, were found on 37% Ficoll. This fraction also contains up to 40% sustentacular cells. A yield of 20% was measured for olfactory cell isolation. Vital staining and ability to synthesize RNA indicate a viability of the final preparation of 70% to 80%. Further identification of the cells was performed by measuring the binding activity of a series of amino acids to a preparation enriched in olfactory cells. A good correlation was determined between the extent of the binding and the reported electrophysiological activities of these amino acids recorded in vivo. Although the final olfactory cell suspension is not pure, it constitutes the first step in the study of the olfactory receptor sites.
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Paul Cancalon (1978) studied this question.