Ruppert Anna-Lena, Keshavarz Maryam, Winterberg Sarah, Oberwinkler Johannes, Kummer Wolfgang, Schütz Burkhard
Institute for Anatomy and Cell Biology, Philipps-University, Robert-Koch-Straße 8, 35037, Marburg, Germany.
Institute for Anatomy and Cell Biology, Justus-Liebig-University, Aulweg 123, 35385, Gießen, Germany.
J Mol Histol. 2020 Aug;51(4):421-435. doi: 10.1007/s10735-020-09893-6. Epub 2020 Jul 2.
Tuft cells are a rare population of chemosensory cells at the mucosal surface epithelia of hollow organs. Their name-giving morphological feature is an apical tuft of stiff microvilli. Accordingly, the actin-binding protein, villin, was identified as one of the first tuft cell markers in immunohistochemical analysis. Unfortunately, villin expression is not restricted to tuft cells, but is also prominent e.g. in enterocytes, which limits the use of this gene as a marker and as an experimental tool to genetically target tuft cells. Here, we report that the villin-related protein, advillin, is a specific tuft cell marker in the gastro-intestinal and biliary tract epithelia. In situ hybridization and immunohistochemistry revealed that advillin expression, unlike villin, was restricted to solitary cholinergic tuft cells in the mucosal linings of the small and large intestine, and in the gall bladder. In the glandular stomach, villin and advillin mRNA were present in all epithelial cells, while detectable protein levels were confined to solitary tuft cells. Advillin expression was no longer detectable in the mucosa of the intestinal and biliary tract from Pou2f3 deficient mice that lack tuft cells. Finally, crossing Avil-Cre transgenic mice with a double-fluorescent reporter mouse line resulted in specific targeting of gastro-intestinal and biliary tuft cells. Our analysis introduces advillin as a selective marker and tool in histological and functional analysis of the alimentary tract tuft cell system.
簇状细胞是存在于中空器官黏膜表面上皮的一种罕见的化学感受细胞群体。其得名的形态学特征是顶端有一束坚硬的微绒毛。因此,肌动蛋白结合蛋白绒毛蛋白在免疫组织化学分析中被鉴定为最早的簇状细胞标志物之一。不幸的是,绒毛蛋白的表达并不局限于簇状细胞,在例如肠上皮细胞中也很显著,这限制了该基因作为簇状细胞标志物以及作为对簇状细胞进行基因靶向的实验工具的应用。在此,我们报告绒毛蛋白相关蛋白促绒毛蛋白是胃肠道和胆道上皮中的一种特异性簇状细胞标志物。原位杂交和免疫组织化学显示,与绒毛蛋白不同,促绒毛蛋白的表达仅限于小肠、大肠黏膜以及胆囊中的孤立胆碱能簇状细胞。在腺胃中,所有上皮细胞都存在绒毛蛋白和促绒毛蛋白的mRNA,而可检测到的蛋白水平仅限于孤立的簇状细胞。在缺乏簇状细胞的Pou2f3基因缺陷小鼠的肠道和胆道黏膜中,促绒毛蛋白的表达不再可检测到。最后,将Avil-Cre转基因小鼠与双荧光报告基因小鼠品系杂交,导致胃肠道和胆道簇状细胞的特异性靶向。我们的分析将促绒毛蛋白引入为消化道簇状细胞系统组织学和功能分析中的一种选择性标志物和工具。