Furue H, Yoshii K
Department of Biochemical Engineering and Science, Kyushu Institute of Technology, Fukuoka, Japan.
J Neurosci Methods. 1998 Oct 1;84(1-2):109-14. doi: 10.1016/s0165-0270(98)00104-6.
In order to investigate taste transduction mechanisms, we developed a method to irrigate the receptor and basolateral membranes of mouse taste bud cells with different solutions under tight-seal recording conditions. A peeled tongue epithelium with taste bud cells was mounted on a recording platform designed to separate irrigating solutions for each membrane. The mucosal surface (receptor membrane side) of the peeled epithelium facing an inner chamber was always irrigated with deionized water or stimulating solutions, and the serosal surface (basolateral membrane side) was irrigated with a physiological saline solution. A recording electrode was placed on the basolateral membrane of a taste bud cell under an upright-microscope with a x 40-water-immersed objective. Investigated taste bud cells generated action potentials, and 1 M glucose, 200 mM NaCl, and 10 mM quinine elicited inward current. Irrigation with deionized water for more than 1 h had no effect. The resistance of the peeled tongue epithelium was 1570 +/- 343 omega cm2. These results show that the peeled tongue epithelium protects basolateral membranes from deionized water or stimulating solutions as the tongue epithelium does in situ and that this method is suitable to investigate the role of each membrane in taste transduction.
为了研究味觉转导机制，我们开发了一种方法，即在紧密密封记录条件下，用不同溶液冲洗小鼠味蕾细胞的受体膜和基底外侧膜。将带有味蕾细胞的剥脱舌上皮安装在一个记录平台上，该平台设计用于分离各膜的冲洗溶液。面向内腔的剥脱上皮的黏膜表面（受体膜侧）始终用去离子水或刺激溶液冲洗，浆膜表面（基底外侧膜侧）用生理盐溶液冲洗。在配备×40水浸物镜的直立显微镜下，将记录电极置于味蕾细胞的基底外侧膜上。所研究的味蕾细胞产生动作电位，1 M葡萄糖、200 mM氯化钠和10 mM奎宁引发内向电流。用去离子水冲洗1小时以上无影响。剥脱舌上皮的电阻为1570±343Ω·cm²。这些结果表明，剥脱舌上皮如同原位舌上皮一样，可保护基底外侧膜免受去离子水或刺激溶液的影响，并且该方法适用于研究各膜在味觉转导中的作用。
