Taste Bud Labeling in Whole Tongue Epithelial Sheet in Adult Mice.
Taste Bud Labeling in Whole Tongue Epithelial Sheet in Adult Mice.
复制标题
成年小鼠整个舌头上皮片的味蕾标记。
DOI：
10.1089/ten.tec.2015.0377
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Liu,Hong-Xiang
中科院分区：
文献类型：
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作者：
Venkatesan,Nandakumar;Boggs,Kristin;Liu,Hong-Xiang
整体组织中的分子标记提供了一种有效的方法来获得关于许多器官和组织的形成、维持、退化和再生的一般信息。然而，在成年小鼠的整个舌组织中标记舌味蕾一直是有问题的，因为舌上皮的强渗透性屏障。在这项研究中，我们提出了一个简单的方法标记味蕾在完整的舌上皮片的成年小鼠。舌内注射蛋白酶孵育后，立即用4%多聚甲醛将舌固定在下颌骨上，可使舌上皮在剥离后保持良好的原位形态。剥离的上皮细胞可用于用泛味细胞标记物角蛋白8和II型味细胞标记物α-味蛋白标记味蕾，在所有三种类型的味乳头中，即菌状、叶状和轮廓状。舌上皮片与一抗和二抗过夜孵育足以用荧光和DAB可视化强烈标记味蕾。标记的单个味蕾易于识别和量化。该方案提供了一种有效的方法，味蕾的表型分析，特别是关于分布模式和数量。
Molecular labeling in whole-mount tissues provides an efficient way to obtain general information about the formation, maintenance, degeneration, and regeneration of many organs and tissues. However, labeling of lingual taste buds in whole tongue tissues in adult mice has been problematic because of the strong permeability barrier of the tongue epithelium. In this study, we present a simple method for labeling taste buds in the intact tongue epithelial sheet of an adult mouse. Following intralingual protease injection and incubation, immediate fixation of the tongue on mandible in 4% paraformaldehyde enabled thein situshape of the tongue epithelium to be well maintained after peeling. The peeled epithelium was accessible to taste bud labeling with a pan-taste cell marker, keratin 8, and a type II taste cell marker, α-gustducin, in all three types of taste papillae, that is, fungiform, foliate, and circumvallate. Overnight incubation of tongue epithelial sheets with primary and secondary antibodies was sufficient for intense labeling of taste buds with both fluorescent and DAB visualizations. Labeled individual taste buds were easy to identify and quantify. This protocol provides an efficient way for phenotypic analyses of taste buds, especially regarding distribution pattern and number.