Regenerative Bioscience Center, Department of Animal and Dairy Science, College of Agricultural and Environmental Sciences, University of Georgia, Athens, GA, USA.
Department of Poultry Sciences, College of Agricultural and Environmental Sciences, University of Georgia, Athens, GA, USA.
Sci Rep. 2016 Nov 17;6:37247. doi: 10.1038/srep37247.
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.
在鸡中，味觉的感觉器官是口腔中的味蕾，通过扫描电子显微镜（SEM）估计，总共有~240-360 个味蕾。目前还没有一种简单的方法可以可视化鸡所有的味蕾。在这里，我们报告了一种使用分子标记物波形蛋白和α-Gustducin 标记鸡口腔上皮片上味觉感受器的高效方法。在用亚上皮注射的蛋白酶孵育后立即进行组织固定，使我们能够剥离整个上皮片，而保持组织的形状和完整性。在剥离的上皮片中，用针对波形蛋白和α-Gustducin 的抗体标记的味蕾在光镜下很容易识别和计数，并且比以前用 SEM 报道的数量更多，呈现出玫瑰花结样簇的模式。与其他组相比，肉鸡型雄性鸡的味蕾更多，并在孵化后的阶段继续增加味蕾的数量。除了卵形的味蕾外，还通过双光子显微镜观察到鸡中存在大的管状味蕾。我们用分子标记物标记味蕾的方案将有助于未来研究鸡味觉感受器的发育及其与摄食行为的关系。
