Yoshimoto Joto, Okada Shinji, Kishi Mikiya, Misaka Takumi
Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan (JY, SO, TM)
Central Research Institute, Mizkan Holdings, Aichi, Japan (JY, MK)
J Histochem Cytochem. 2016 Mar;64(3):205-15. doi: 10.1369/0022155415626987. Epub 2015 Dec 30.
Taste signals are received by taste buds. To better understand the taste reception system, expression patterns of taste-related molecules are determined by in situ hybridization (ISH) analyses at the histological level. Nevertheless, even though ISH is essential for determining mRNA expression, few taste bud markers can be applied together with ISH. Ulex europaeus agglutinin-1 (UEA-1) appears to be a reliable murine taste bud marker based on immunohistochemistry (IHC) analyses. However, there is no evidence as to whether UEA-1 can be used for ISH. Thus, the present study evaluated UEA-1 using various histochemical methods, especially ISH. When lectin staining was performed after ISH procedures, UEA-1 clearly labeled taste cellular membranes and distinctly indicated boundaries between taste buds and the surrounding epithelial cells. Additionally, UEA-1 was determined as a taste bud marker not only when used in single-colored ISH but also when employed with double-labeled ISH or during simultaneous detection using IHC and ISH methods. These results suggest that UEA-1 is a useful marker when conducting analyses based on ISH methods. To clarify UEA-1 staining details, multi-fluorescent IHC (together with UEA-1 staining) was examined, resulting in more than 99% of cells being labeled by UEA-1 and overlapping with KCNQ1-expressing cells.
味觉信号由味蕾接收。为了更好地理解味觉接收系统,在组织学水平上通过原位杂交(ISH)分析来确定味觉相关分子的表达模式。然而,尽管ISH对于确定mRNA表达至关重要,但很少有味蕾标记物能与ISH一起应用。基于免疫组织化学(IHC)分析,欧洲荆豆凝集素-1(UEA-1)似乎是一种可靠的小鼠味蕾标记物。然而,尚无证据表明UEA-1是否可用于ISH。因此,本研究使用各种组织化学方法,尤其是ISH,对UEA-1进行了评估。在ISH程序后进行凝集素染色时,UEA-1清晰地标记了味觉细胞膜,并明确显示了味蕾与周围上皮细胞之间的边界。此外,UEA-1不仅在单色ISH中使用时被确定为味蕾标记物,而且在双标记ISH中使用或在IHC和ISH方法同时检测期间也被确定为味蕾标记物。这些结果表明,在基于ISH方法进行分析时,UEA-1是一种有用的标记物。为了阐明UEA-1染色细节,对多重荧光IHC(连同UEA-1染色)进行了检查,结果超过99%的细胞被UEA-1标记,并与表达KCNQ1的细胞重叠。