Abstract
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.
Highlights
In chickens, molecular markers have been used to label taste buds in tissue sections[7,8,9]
Taste bud labeling with the molecular markers Vimentin and α-Gustducin was successful in peeled epithelial sheets from the base of the oral cavity, the palate, and the posterior region of the tongue, areas where taste buds are located
We found that antibodies to molecular markers present in the taste buds labeled many more taste buds than previously counted using conventional scanning electron microscopy (SEM) methods
Summary
Molecular markers have been used to label taste buds in tissue sections[7,8,9]. Previous studies have demonstrated that Vimentin and α-Gustducin each label a subpopulation of taste buds in chicken[8,9]. Each molecular marker labeled only a subpopulation of taste bud cells in post-hatch chickens, both markers were observed in all the taste buds examined. Α-Gustducin and Vimentin are potentially useful molecular markers for visualizing chicken taste buds in whole mount tissue. In the present study we tested a protocol for peeling off intact oral epithelial sheets in post-hatch broiler-type chickens. Taste bud labeling with the molecular markers Vimentin and α-Gustducin was successful in peeled epithelial sheets from the base of the oral cavity, the palate, and the posterior region of the tongue, areas where taste buds are located. The protocol may have broad application across fields, i.e., it is applicable to all epithelial appendages in different species
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