Kan Hideko, Kataoka-Shirasugi Naoko, Amakawa Taisaku
Department of Human Environmental Science, Graduate School of Human Development and Environment, Kobe University, 3-11 Tsurukabuto, Nada 657-8501, Japan.
J Insect Physiol. 2008 Jun;54(6):1028-34. doi: 10.1016/j.jinsphys.2008.04.002. Epub 2008 Apr 11.
The whole cell clamp method was directly applied to the sensory receptor neurons isolated from the adult labellar hair of the blow fly Phormia regina to locate the signal transduction pathways mediated by second messengers. First, the cAMP-mediated transduction pathway was examined to specify its location in the sugar receptor cell. Sugar receptor cell was identified by recording inward current flow under the voltage clamp applying sucrose solution to the surface of the taste neurons. When cyclic nucleotides, such as cGMP and cAMP, were introduced into the sugar receptor cell, inward current was observed (cGMP, 70pA; cAMP, 300pA at 350microM). Inhibitors and activators for the second messengers (GDPbetaS and forskolin) and non-cyclic nucleotides were also examined. Second, non-nucleotide second messengers (IP3 and Ca2+) were examined. The sugar receptor cell was activated when it was injected with IP3 or Ca2+. All the obtained results suggest that the cAMP-mediated signal transduction pathway plays a major role in the sugar receptor cell. The possibility of other transduction pathways mediated by IP3 or Ca2+ was not excluded.
全细胞钳制方法直接应用于从成年丽蝇(Phormia regina)唇须毛中分离出的感觉受体神经元,以定位由第二信使介导的信号转导途径。首先,检测cAMP介导的转导途径,以确定其在糖受体细胞中的位置。通过在电压钳制下向味觉神经元表面施加蔗糖溶液记录内向电流来鉴定糖受体细胞。当将环核苷酸,如cGMP和cAMP,引入糖受体细胞时,观察到内向电流(cGMP,70pA;350μM时cAMP为300pA)。还检测了第二信使的抑制剂和激活剂(GDPβS和福斯可林)以及非环核苷酸。其次,检测了非核苷酸第二信使(IP3和Ca2+)。当向糖受体细胞注射IP3或Ca2+时,该细胞被激活。所有获得的结果表明,cAMP介导的信号转导途径在糖受体细胞中起主要作用。不排除由IP3或Ca2+介导的其他转导途径的可能性。