Department of Endodontics and Clinical Cardiology, Tokyo Dental College, Japan; Department of Pathology, Tokyo Dental College, Japan; Oral Health Science Center hrc8, Tokyo Dental College, Japan.
Microsc Res Tech. 2013 Oct;76(10):988-91. doi: 10.1002/jemt.22271. Epub 2013 Aug 6.
Gap junctions play an important role in differentiation of odontoblasts. Gap junction protein, connexin 43 is expressed in odontoblast. However, the detailed localization in odontoblasts has yet to be fully investigated. We investigated the localization of connexin43 in rat odontoblasts immuno-electron microscopically. The rats were transcardially fixed with 1% paraformaldehyde in 0.1M phosphate buffer, and mandibles were decalcified with 10% ethylenediamine tetraacetic acid. Pre-embedding method was carried out for immuno-electron microscopic analysis. Microscopically, gap junctions were localized between bodies of odontoblasts, and between bodies and processes of odontoblasts. The gap junctions were labeled with gold particles that indicated connexin43. These results suggest that gap junctions between odontoblasts are definitely composed of connexin43 in rats, and our methods used in this study is useful to investigate localization of connexin43 immuno-electron microscopically.
缝隙连接在成牙本质细胞分化中发挥重要作用。缝隙连接蛋白连接蛋白 43 在成牙本质细胞中表达。然而，其在成牙本质细胞中的详细定位尚未完全研究清楚。本研究通过免疫电镜观察大鼠成牙本质细胞中连接蛋白 43 的定位。用 1%多聚甲醛和 0.1M 磷酸缓冲液心脏灌注固定大鼠，用 10%乙二胺四乙酸脱钙。进行预包埋免疫电镜分析。镜下观察到成牙本质细胞体之间以及成牙本质细胞体与突起之间存在缝隙连接。缝隙连接用标记连接蛋白 43 的金颗粒标记。这些结果表明，大鼠成牙本质细胞之间的缝隙连接确实由连接蛋白 43 组成，本研究中使用的方法可用于免疫电镜观察连接蛋白 43 的定位。
