Ulex Europaeus Agglutinin-1 Is a Reliable Taste Bud Marker for In Situ Hybridization Analyses
Ulex Europaeus Agglutinin-1 Is a Reliable Taste Bud Marker for In Situ Hybridization Analyses
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DOI：
10.1369/0022155415626987
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发表时间：
2016-03-01
影响因子：
3.2
通讯作者：
Misaka, Takumi
中科院分区：
文献类型：
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作者：
Yoshimoto, Joto;Okada, Shinji;Misaka, Takumi
味觉信号由味蕾接收。为了更好地了解味觉接受系统，味觉相关分子的表达模式通过原位杂交（ISH）分析在组织学水平上确定。然而，尽管ISH是确定mRNA表达所必需的，但很少有味蕾标记物可以与ISH一起应用。荆豆凝集素-1（UEA-1）似乎是一个可靠的基于免疫组织化学（IHC）分析的小鼠味蕾标志物。然而，没有证据表明UEA-1是否可以用于ISH。因此，本研究使用各种组织化学方法，特别是ISH来评估UEA-1。当ISH程序后进行凝集素染色时，UEA-1清楚地标记了味觉细胞膜，并清楚地指示了味蕾与周围上皮细胞之间的边界。此外，UEA-1被确定为味蕾标记物，不仅在单色ISH中使用时，而且在与双标记ISH一起使用时或在使用IHC和ISH方法同时检测期间使用时。这些结果表明，UEA-1是一个有用的标志物进行分析的基础上ISH方法。为了阐明UEA-1染色细节，检查了多荧光IHC（与UEA-1染色一起），结果超过99%的细胞被UEA-1标记，并与KCNQ 1表达细胞重叠。
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.