Biochemical studies of taste sensation. III. Preparation of a suspension of bovine taste bud cells and their labeling with a fluorescent probe.
A method to prepare suspensions of taste bud cells is described. Bovine circumvallate papillae, which contain most of the taste buds in this animal, are incubated in collagenase-containing medium and the epidermal sidewall tissue is then dissected from the inner gelatinous dermis. The sidewall tissue, which contains the taste buds, is gently homogenized by manual operation of an all-glass homogenizer with a loose-fitting pestle. The suspended material is separated on a discontinous Ficoll gradient (2%, 8%, 10%, 12% w/w). The material banding at the 8-2% interface is greatly enriched in spindle-shaped cells that are morphologically similar to taste bud cells as they appear in situ. These cells are not seen when the procedure is done with tissues devoid of taste buds, namely the upper surface of the circumvallate papilla or epithelium from the intermolar eminence. Fluorescence analysis indicates that the hydrophobic probe, 8-anilino-1-naphthalenesulfonate (ANS), binds to relatively nonpolar sites in the suspension. It is postulated that the probe is adsorbing onto the surface membrane of the cell. These preparations may be useful in studying specificity and transduction in taste sensation.
D000817:Anilino Naphthalenesulfonates / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002499:Centrifugation, Density Gradient; D005453:Fluorescence; D005615:Freezing; D006652:Histological Techniques; D066298:In Vitro Techniques; D003012:Microbial Collagenase / Q000378:metabolism; D010101:Oxygen Consumption; D011955:Receptors, Drug / Q000187:drug effects; D013649:Taste / Q000502:physiology*; D013650:Taste Buds / Q000166:cytology / Q000378:metabolism*
D000817:Anilino Naphthalenesulfonates; D011955:Receptors, Drug; D003012:Microbial Collagenase