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In mammals, each cell group within the taste buds expresses either the T1R or the T2R taste receptor for preference-aversion discrimination. However, no such information is available regarding fish.

We developed a novel system for quantitatively assaying taste preference-aversion in medaka fish. In this study, we prepared fluorescently labeled foods with fine cavities designed to retain tastants until they were bitten by the fish.

The subjects were fed food containing a mixture of amino acids and inosine monophosphate AN food , denatonium benzoate DN food or no tastant NT food , and the amounts of ingested food were measured by fluorescence microscopy.

Statistical analysis of the fluorescence intensities yielded quantitative measurements of AN food preference and DN food aversion.

We then generated a transgenic fish expressing dominant-negative G alpha i2 both in T1R-expressing and in T2R-expressing cells.

The feeding assay revealed that the transgenic fish was unable to show a preference for AN food and an aversion to DN food.

The assay system was useful for evaluating taste-blind behaviors, and the results indicate that the two taste signaling pathways conveying preferable and aversive taste information are conserved in fish as well as in mammals.

Using a transgeric zebrafish line that expressed green fluorescent protein under the control of the PLC-beta 2 promoter, we showed that zfTRPM5 is expressed in green fluorescent protein-labeled cells of the taste buds.

These results demonstrate that zfTRPM5 and PLC-beta 2 colocalize in zebrafish taste receptor cells, suggesting their crucial roles in taste signaling via the fish taste receptors.

Similar taste receptors have also been characterized in fish, but their ligands have not been identified yet.

In the present study, we conducted a series of experiments to identify the fish taste receptor ligands. Facial nerve recordings in zebrafish Danio rerio demonstrated that the fish perceived amino acids and even denatonium, which is a representative of aversive bitter compounds for mammals and Drosophila.

Behavior observations confirmed that zebrafish prefer amino acids and avoid denatonium. These results suggest that, although there may be some fish-specific way of discriminating ligands, vertebrates could have a conserved gustatory mechanism by which T1Rs and T2Rs respond to attractive and aversive tastants, respectively.

Vertebrate taste receptor cells express signaling molecules such as taste receptors and effectors to convert taste stimuli to inner cellular signals.

Phospholipase C-beta 2 PLC-beta 2 is an effector enzyme that is necessary to transduce taste signals in the mouse.

It was shown that a subset of the plc-beta 2 expressing cells also express taste receptor molecules, T1Rs or T2Rs, in mammals and fish.

To label plc-beta 2 expressing cells in the model fish species, we constructed a transgene by linking the 5'-upstream region of the medaka plc-beta 2 gene to a green fluorescent protein GFP gene.

The resulting transgenic medaka exhibited GFP signals in taste buds of the lips and the pharyngeal region. Detailed observation revealed that the GFP signals were in a subpopulation of taste bud cells, and co-localized with the transcript of endogenous plc-beta 2 gene.

Zebrafish introduced with the same transgene showed GFP signals in a subpopulation of taste bud cells of the lips and the pharyngeal region as in the case of medaka.

This is the first report of successful labeling of taste receptor cells in two model fish species under the control of the plc-beta 2 promoter.

This promoter will be a useful genetic tool to study the vertebrate taste system in general. All rights reserved.

Phospholipase C-beta 2 PLC-beta 2 is a key enzyme in mammalian taste signal transduction. To analyze the taste system in fishes at molecular level, we cloned mfplc-beta 2 as a medaka fish homologue of PLC-beta 2.

In situ hybridization analysis revealed that mfplc-beta 2 is expressed in the lip and branchial region where chemosensory tissues are distributed.

Immunohistochemical detection of nerve fibers near the mfplc-beta 2 positive cells suggests the characteristic of peripheral sensory cells. These results suggest that mfplc-beta 2 is expressed in the gustatory sensory cells of medaka.

This may provide a molecular basis for the taste reception at multiple tissues in fish species. C Elsevier Ireland Ltd. We performed a systematic screen for mutations affecting the trajectory of axons visualized by immunohistochemical staining of Medaka embryos with anti-acetylated tubulin antibody.

Among the mutations identified, yanagi yan and kazura kaz mutations caused specific defects in projection of the posterior lateral line PLL nerve.

In yan and kaz mutant embryos, the PLL nerve main bundle was misrouted ventrally and dorsally or anteriorly. Medaka semaphorin3A, sdf1, and cxcr4 cDNA fragments were cloned to allow analysis of these mutants.

There were no changes in semaphorin3A or sdf1 expression in mutant embryos, suggesting that the tissues expressing semaphorin3A or sqf1 that are involved in PLL nerve guidance are present in these mutant embryos.

Double staining revealed that the mislocated PLL primordium and growth cone of the ectopically projected PLL nerve were always colocalized in both yan and kaz mutant embryos, suggesting that migration of PLL primordia and PLL nerve growth cones are not uncoupled in these mutants.

Although homozygous yan larvae showed incomplete migration of the PLL primordium along the anteroposterior axis, ventral proneuromast migration was complete, suggesting that ventral migration of the proneuromast does not require the signaling affected in yan mutants.

In addition to the PLL system, the distribution of primordial germ cells PGCs was also affected in both yan and kaz mutant embryos, indicating that yan and kaz genes are required for the migration of both PLL primordia and PGCs.

Genetic linkage analysis indicated that kaz is linked to cxcr4, but yan is not linked to sdf1 or cxcr4. These mutations will provide genetic clues to investigate the molecular mechanism underlying formation of the PLL system.

Research Activities Profile and Settings Profile My interests are: - Mechanisms underlying the perception of food compounds in our body.

Research Interests apetite. Lipid peroxides as endogenous oxidants forming 8-oxo-guanosine and lipid-soluble antioxidants as suppressing agents.

The combined effect of clothianidin and environmental stress on the behavioral and reproductive function in male mice. Construction of a taste-blind medaka fish and quantitative assay of its preference-aversion behavior.

Transient receptor potential channel M5 and phospholipaseC-beta 2 colocalizing in zebrafish taste receptor cells. Citation: Nichani, V.

Download Citation. Affiliated: Simerics Inc. Related Topics: Computational fluid dynamics Engine lubrication systems Engine lubricants.

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