Isolation of chicken taste buds for real‐time <scp><scp>Ca<sup>2</sup></scp></scp><sup>+</sup> imaging

Kudo, Ken Ichi; Kawabata, Fuminori; Nomura, Toumi; Aridome, Ayumi; Nishimura, Shigeyuki; Shoji, Tetsuo

doi:10.1111/asj.12222

Cited by 18 publications

(16 citation statements)

References 33 publications

(33 reference statements)

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“…In rats, taste bud cells that express α-Gustducin have a long life span compared to other cell types [27]. α -Gustducin is explicitly expressed in mature chicken taste buds at P3 [9] and is highly expressed in an isolated chicken taste bud-like subset of cells [30]. In the present study, we detected α-Gustducin immunosignals in immature taste buds as early as E18.…”

Section: Discussionsupporting

confidence: 50%

Distribution of α-Gustducin and Vimentin in premature and mature taste buds in chickens

Venkatesan

Rajapaksha

Payne

et al. 2016

Biochemical and Biophysical Research Communications

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The sensory organs for taste in chickens (Gallus sp.) are taste buds in the oral epithelium of the palate, base of the oral cavity, and posterior tongue. Although there is not a pan-taste cell marker that labels all chicken taste bud cells, α-Gustducin and Vimentin each label a subpopulation of taste bud cells. In the present study, we used both α-Gustducin and Vimentin to further characterize chicken taste buds at the embryonic and post-hatching stages (E17-P5). We found that both α-Gustducin and Vimentin label distinct and overlapping populations of, but not all, taste bud cells. A-Gustducin immunosignals were observed as early as E18 and were consistently distributed in early and mature taste buds in embryos and hatchlings. Vimentin immunoreactivity was initially sparse at the embryonic stages then became apparent in taste buds after hatch. In hatchlings, α-Gustducin and Vimentin immunosignals largely co-localized in taste buds. A small subset of taste bud cells were labeled by either α-Gustducin or Vimentin or were not labeled. Importantly, each of the markers was observed in all of the examined taste buds. Our data suggest that the early onset of α-Gustducin in taste buds might be important for enabling chickens to respond to taste stimuli immediately after hatch and that distinctive population of taste bud cells that are labeled by different molecular markers might represent different cell types or different phases of taste bud cells. Additionally, α-Gustducin and Vimentin can potentially be used as molecular markers of all chicken taste buds in whole mount tissue.

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Section: Discussionsupporting

confidence: 50%

Distribution of α-Gustducin and Vimentin in premature and mature taste buds in chickens

Venkatesan

Rajapaksha

Payne

et al. 2016

Biochemical and Biophysical Research Communications

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“…Chickens show aversive responses to bitter and sour (Ganchrow et al 1990;Kudo et al 2010a;Hirose et al 2015;Dey et al 2017) and a preference for feeds containing fat and umami tastants (Sawamura et al 2015;Yoshida et al 2015), and we demonstrated that isolated taste bud cells of chickens responded to umami and bitter solutions (Kudo et al 2014). These findings indicate that the taste sense of chickens is similar to that of mammals.…”

Section: Introductionmentioning

confidence: 53%

Expression levels of taste‐related genes in palate and tongue tip, and involvement of transient receptor potential subfamily M member 5 (TRPM5) in taste sense in chickens

Yoshida

Kawabata

et al. 2017

Animal Science Journal

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The elucidation of the mechanisms underlying the taste sense of chickens will contribute to improvements in poultry feeding, because the molecular mechanism of chickens' taste sense defines the feeding behavior of chickens. Here we focused on the gene expressions in two different oral tissues of chickens - the palate, which contains many taste buds, and the tongue tip, which contains few taste buds. Using the quantitative real-time polymerase chain reaction method, we found that the molecular markers for taste buds of chickens, that is α-gustducin and vimentin, were expressed significantly highly in the palate compared to the tongue tip. Our analyses also revealed that transient receptor potential subfamily M member 5 (TRPM5), a cation channel involved in taste transduction in mammals, was also highly expressed in the palate compared to the tongue tip. Our findings demonstrated that the expression patterns of these genes were significantly correlated. We showed that the aversion to bitter solution was alleviated by a TRPM5 inhibitor in behavior of chickens. Taken together, our findings enabled us to develop a simple method for screening taste-related genes in chickens. The use of this method demonstrated that TRPM5 was involved in chickens' taste transduction, and that a TRPM5 inhibitor can alleviate chickens' bitter taste perception of feed ingredients.

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“…It has been reported that α-Gustducin is specifically expressed in chicken taste buds911. Two molecular markers, Vimentin and α-Gustducin, were used to label taste buds in the epithelial sheets of the base of the oral cavity in P0 to P8 chickens.…”

Section: Resultsmentioning

confidence: 99%

Labeling and analysis of chicken taste buds using molecular markers in oral epithelial sheets

Rajapaksha

Wang

Venkatesan

et al. 2016

Sci Rep

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In chickens, the sensory organs for taste are the taste buds in the oral cavity, of which there are ~240–360 in total number as estimated by scanning electron microscopy (SEM). There is not an easy way to visualize all taste buds in chickens. Here, we report a highly efficient method for labeling chicken taste buds in oral epithelial sheets using the molecular markers Vimentin and α-Gustducin. Immediate tissue fixation following incubation with sub-epithelially injected proteases enabled us to peel off whole epithelial sheets, leaving the shape and integrity of the tissue intact. In the peeled epithelial sheets, taste buds labeled with antibodies against Vimentin and α-Gustducin were easily identified and counted under a light microscope and many more taste buds, patterned in rosette-like clusters, were found than previously reported with SEM. Broiler-type, female-line males have more taste buds than other groups and continue to increase the number of taste buds over stages after hatch. In addition to ovoid-shaped taste buds, big tube-shaped taste buds were observed in the chicken using 2-photon microscopy. Our protocol for labeling taste buds with molecular markers will factilitate future mechanistic studies on the development of chicken taste buds in association with their feeding behaviors.

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Isolation of chicken taste buds for real‐time Ca²⁺ imaging

Cited by 18 publications

References 33 publications

Distribution of α-Gustducin and Vimentin in premature and mature taste buds in chickens

Distribution of α-Gustducin and Vimentin in premature and mature taste buds in chickens

Expression levels of taste‐related genes in palate and tongue tip, and involvement of transient receptor potential subfamily M member 5 (TRPM5) in taste sense in chickens

Labeling and analysis of chicken taste buds using molecular markers in oral epithelial sheets

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Isolation of chicken taste buds for real‐time Ca2+ imaging

Cited by 18 publications

References 33 publications

Distribution of α-Gustducin and Vimentin in premature and mature taste buds in chickens

Distribution of α-Gustducin and Vimentin in premature and mature taste buds in chickens

Expression levels of taste‐related genes in palate and tongue tip, and involvement of transient receptor potential subfamily M member 5 (TRPM5) in taste sense in chickens

Labeling and analysis of chicken taste buds using molecular markers in oral epithelial sheets

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Isolation of chicken taste buds for real‐time Ca²⁺ imaging