Disc Electrophoresis of Extracts from the Taste Buds Located in Circumvallate Papillae of Rat Tongues

Uehara, Seiji

doi:10.1085/jgp.61.3.290

Cited by 11 publications

(8 citation statements)

References 15 publications

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“…In addition to the failure of the other groups, in direct experimental tests, to substantiate the earlier claims, Koyama and Kurihara (44) showed that the "sweet-sensitive protein" is present in large amounts throughout the bovine tongue epithelium whether or not taste buds are present. Later reports show that unique proteins do exist in preparations derived from taste papillae and taste bud cells (45,46), but their function has yet to be elucidated.…”

Section: Methodsmentioning

confidence: 99%

Biochemical studies of taste sensation: binding to taste tissue of 3H-labeled monellin, a sweet-tasting protein.

Çağan¹,

Morris²

1979

Proc. Natl. Acad. Sci. U.S.A.

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Binding of 3H-labeled methylated monellin to taste receptor tissue was demonstrated in vitro. Preparations of bovine and human circumvallate (taste) papillae bound more of the ligand than did lingual and nonlingual epithelial preparations devoid of taste buds. Binding to the taste preparations saturated at high ligand concentrations. Furthermore, sugars and other sweet-tasting molecules appeared to compete to some extent with this sweet-tasting protein for its binding sites. These binding measurements of the intensely sweet-tasting protein monellin to taste receptor preparations help to establish the binding interaction as an initial step in taste sensation.Relatively little is understood about the biochemical basis of sweet taste despite considerable knowledge of a wide variety of sweet-tasting chemical compounds (1)(2)(3)(4)(5)(6). Most of the behavioral, physiological, and biochemical research on sweet taste has used sucrose or other sugars. The inherently weak binding (6-8) of sugars to taste receptors, with values of KD in the range of 10-1 to 10-3 M (9, 10), presents a substantial difficulty in directly studying the biochemical basis of how a sugar binds to and triggers a taste receptor to respond. The interest in our laboratory (11)(12)(13)(14)(15)(16) formaldehyde was diluted with unlabeled formaldehyde to yield 2.7 M and a specific radioactivity of 9.5 Ci/ mol). This was followed by reduction with 100 ,ul of 1.2 M NaBH4 (in water) for 5 min and then with 400,l for another 5 min. The preparation was washed at room temperature with 10 mM sodium phosphate buffer ( To prepare the tissue fraction, we thawed the frozen tongues or human tissue samples and dissected the circumvallate papillae free, with the top surface (0.2-0.5 mm) having been removed with a scalpel. Small (5 X 5 mm) blocks of tongue epithelium provided control tissue devoid of taste buds (bovine and human), and pectoral skin (human) was also used as a control tissue where indicated. The epidermal sidewalls of the papillae and the upper layer from the control epithelial blocks were teased off with forceps (18,19) PA 19013. 1692 The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.

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Section: Methodsmentioning

confidence: 99%

Biochemical studies of taste sensation: binding to taste tissue of 3H-labeled monellin, a sweet-tasting protein.

Çağan¹,

Morris²

1979

Proc. Natl. Acad. Sci. U.S.A.

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“…The procedure reported for isolating rat circumvallate taste buds (Uehara, 1973) differs substantially from our procedure. The rat papillae were digested in a medium containing 0.005% elastase and 0.08% trypsin for a total of 12 hr.…”

Section: Use Of Collagenasementioning

confidence: 98%

“…The epidermal material was collected from the papillae and the taste buds were then excised by dissection with the aid of a dissecting microscope. In the procedure of Uehara (1973), it was intended to isolate whole taste buds, rather than the cells themselves as in the present method. a "Suspended" protein is the amount that was released from the tissue by treatment with a loose-fitting all-glass homogenizer as described in Methods.…”

Section: Use Of Collagenasementioning

confidence: 99%

“…Further studies will be required to define the physiological characteristics of the isolated cells. No information is available on the viability of the isolated rat taste buds (Uehara, 1973).…”

Section: Respiration Measurementsmentioning

confidence: 99%

“…A preliminary report of this work was published (Brand and Cagan, 1973). During the course of these studies a microscale procedure for isolating whole rat taste buds, rather than taste bud cells as in the present work, was reported (Uehara, 1973).…”

Section: Introductionmentioning

confidence: 96%

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Biochemical studies of taste sensation. III. Preparation of a suspension of bovine taste bud cells and their labeling with a fluorescent probe

Brand

Çağan

1976

J. Neurobiol.

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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.

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The negatively charged protein extracted from Tetrahymena pyriformis as an attractant in Amoeba proteus chemotaxis

Nohmi

Tawada

1974

Journal Cellular Physiology

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Disc Electrophoresis of Extracts from the Taste Buds Located in Circumvallate Papillae of Rat Tongues

Cited by 11 publications

References 15 publications

Biochemical studies of taste sensation: binding to taste tissue of 3H-labeled monellin, a sweet-tasting protein.

Biochemical studies of taste sensation: binding to taste tissue of 3H-labeled monellin, a sweet-tasting protein.

Biochemical studies of taste sensation. III. Preparation of a suspension of bovine taste bud cells and their labeling with a fluorescent probe

The negatively charged protein extracted from Tetrahymena pyriformis as an attractant in Amoeba proteus chemotaxis

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