TMEM16A,跨膜蛋白16家族的成员,作为钙激活的氯化物通道(CaCC)的分子基础。我们使用RT-PCR证明TMEM16A在棉铃虫神经元中的表达,并首次使用膜片钳技术记录了棉铃虫急性分离神经元的CaCCs电流。为了筛选钙激活氯离子通道的有效抑制剂,四种氯通道抑制剂的抑制作用,CaCCinh-A01,NPPB,DIDS,和SITS,对CaCC进行了比较。四种抑制剂对CaCCs外向电流的抑制作用为CaCCinh-A01(10μM,56.31),NPPB(200μM,43.69%),SITS(1mM,12.41%)和DIDS(1mM,13.29%)。在这些抑制剂中,CaCCinh-A01表现出作为阻断剂的最高功效。为了进一步探索钙通道蛋白是否可以作为拟除虫菊酯的潜在靶标,我们比较了(I型)替氟菊酯和(II型)溴氰菊酯对CaCCs的影响。10μM和100μM的替氟菊酯可以刺激CaCC中的大的尾电流,将其停用时间延长10.44ms和31.49ms,并且V0.5在超极化中偏移2-8mV。然后,溴氰菊酯对CaCCs的失活和活化无明显影响。因此,棉铃虫CaCC可作为拟除虫菊酯的潜在靶标,但Ⅰ型和Ⅱ型拟除虫菊酯对CaCCs的影响不同。
TMEM16A, a member of the Transmembrane protein 16 family, serves as the molecular basis for calcium activated chloride channels (CaCCs). We use RT-PCR to demonstrate the expression of TMEM16A in the neurons of Helicoverpa armigera, and records the CaCCs current of acute isolated neurons of H. armigera for the first time using patch clamp technology. In order to screen effective inhibitors of calcium-activated chloride channels, the inhibitory effects of four chloride channel inhibitors, CaCCinh-A01, NPPB, DIDS, and SITS, on CaCCs were compared. The inhibitory effects of the four inhibitors on the outward current of CaCCs were CaCCinh-A01 (10 μM, 56.31), NPPB (200 μM, 43.69 %), SITS (1 mM, 12.41 %) and DIDS (1 mM, 13.29 %). Among these inhibitors, CaCCinh-A01 demonstrated the highest efficacy as a blocker. To further explore whether calcium channel proteins can serve as potential targets of pyrethroids, we compared the effects of (type I) tefluthrin and (type II) deltamethrin on CaCCs. 10 μM and 100 μM tefluthrin can stimulate a large tail current in CaCCs, prolonging their deactivation time by 10.44 ms and 31.49 ms, and the V0.5 shifted in the hyperpolarization by 2-8 mV. Then, deltamethrin had no obvious effect on the deactivation and activation of CaCCs. Therefore, CaCCs of H. armigera can be used as a potential target of pyrethroids, but type I and type II pyrethroids have different effects on CaCCs.