Immunohistochemical detection of TAS2R38 protein in human taste cells

The sense of taste plays an important role in the evaluation of the nutrient composition of consumed food. Bitter taste in particular is believed to serve a warning function against the ingestion of poisonous substances. In the past years enormous progress was made in the characterization of bitter...

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Veröffentlicht in:PloS one 2012-07, Vol.7 (7), p.e40304
Hauptverfasser: Behrens, Maik, Born, Stephan, Redel, Ulrike, Voigt, Nadine, Schuh, Vanessa, Raguse, Jan-Dirk, Meyerhof, Wolfgang
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container_start_page e40304
container_title PloS one
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creator Behrens, Maik
Born, Stephan
Redel, Ulrike
Voigt, Nadine
Schuh, Vanessa
Raguse, Jan-Dirk
Meyerhof, Wolfgang
description The sense of taste plays an important role in the evaluation of the nutrient composition of consumed food. Bitter taste in particular is believed to serve a warning function against the ingestion of poisonous substances. In the past years enormous progress was made in the characterization of bitter taste receptors, including their gene expression patterns, pharmacological features and presumed physiological roles in gustatory as well as in non-gustatory tissues. However, due to a lack in TAS2R-specifc antibodies the localization of receptor proteins within gustatory tissues has never been analyzed. In the present study we have screened a panel of commercially available antisera raised against human bitter taste receptors by immunocytochemical experiments. One of these antisera was found to be highly specific for the human bitter taste receptor TAS2R38. We further demonstrate that this antibody is able to detect heterologously expressed TAS2R38 protein on Western blots. The antiserum is, however, not able to interfere significantly with TAS2R38 function in cell based calcium imaging analyses. Most importantly, we were able to demonstrate the presence of TAS2R38 protein in human gustatory papillae. Using double immunofluorescence we show that TAS2R38-positive cells form a subpopulation of PLCbeta2 expressing cells. On a subcellular level the localization of this bitter taste receptor is neither restricted to the cell surface nor particularly enriched at the level of the microvilli protruding into the pore region of the taste buds, but rather evenly distributed over the entire cell body.
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Bitter taste in particular is believed to serve a warning function against the ingestion of poisonous substances. In the past years enormous progress was made in the characterization of bitter taste receptors, including their gene expression patterns, pharmacological features and presumed physiological roles in gustatory as well as in non-gustatory tissues. However, due to a lack in TAS2R-specifc antibodies the localization of receptor proteins within gustatory tissues has never been analyzed. In the present study we have screened a panel of commercially available antisera raised against human bitter taste receptors by immunocytochemical experiments. One of these antisera was found to be highly specific for the human bitter taste receptor TAS2R38. We further demonstrate that this antibody is able to detect heterologously expressed TAS2R38 protein on Western blots. The antiserum is, however, not able to interfere significantly with TAS2R38 function in cell based calcium imaging analyses. Most importantly, we were able to demonstrate the presence of TAS2R38 protein in human gustatory papillae. Using double immunofluorescence we show that TAS2R38-positive cells form a subpopulation of PLCbeta2 expressing cells. 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subjects Amino Acid Sequence
Animals
Antibodies
Antisera
Biology
Bitter taste
Blotting, Western
Calcium
Calcium imaging
Calcium Signaling
Cell body
Cell surface
Cytochemistry
Endoplasmic reticulum
Food composition
Gene expression
HEK293 Cells
Humans
Immune Sera - chemistry
Immunofluorescence
Immunoglobulins
Immunohistochemistry
Ingestion
Kinases
Localization
Mammals
Medicine
Mice
Microscopy, Fluorescence
Molecular Sequence Data
Nutrition
Papillae
Pharmacology
Proteins
Receptors
Receptors, G-Protein-Coupled - immunology
Receptors, G-Protein-Coupled - metabolism
Rodents
Taste
Taste (Sense)
Taste buds
Taste Buds - cytology
Taste Buds - metabolism
Taste receptors
Tissues
Tongue
Western blotting
title Immunohistochemical detection of TAS2R38 protein in human taste cells
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