Pertussis toxin （FIX） inhibits the activation of the α-subunit of the inhibitory heterotrimeric G-proteins （Cαi/o） and modulates voltage-gated sodium channels, which may be one of the primary targets of pyrethro...Pertussis toxin （FIX） inhibits the activation of the α-subunit of the inhibitory heterotrimeric G-proteins （Cαi/o） and modulates voltage-gated sodium channels, which may be one of the primary targets of pyrethroids. To investigate the potential mechanisms of agricultural pests resistance to pyrethroid insecticides, we examined the modulations by PTX on sodium channels in the central neurons of the 3rd-4th instar larvae of cyhalothrin-resistant （Cy-R） and cyhaiothrin-susceptible （Cy-S） Helicoverpa armigera by the whole-cell patch-clamp technique. The isolated neurons were cultured for 12-16 h in an improved L15 insect culture medium with or without PTX （400 ng/mL）. The results showed that both the Cy-R and Cy-S sodium channels exhibited fast kinetics and tetrodotoxin （TTX） sensitivity. The Cy-R sodium channels exhibited not only altered gating properties, including a 8.88-mV right shift in voltage-dependent activation （V0.5act） and a 6.54-mV right shift in voltage-dependent inactivation （V0.5inact）, but also a reduced peak in sodium channel density （Ⅰdensity） （55.2% of that in Cy-S neurons）. Cy-R sodium channels also showed low excitability, as evidenced by right shift of activation potential （Ⅴacti） by 5-10 mV and peak potential （Ⅴpcak） by 20 mV. FIX exerted significant effects on Cy-S sodium channels, reducing sodium channel density by 70.04%, right shifting V0.5act by 14.41 mV and V0.5inact by 9. 38 mV. It did not cause any significant changes of the parameters mentioned above in the Cy-R sodium channels. The activation time （Tpeak） from latency to peak at peak voltage and the fast inactivation time constant （τinact） in both Cy-S and Cy-R neurons were not affected. The results suggest that cotton bollworm resistant to pyrethroid insecticides involves not only mutations and allosteric alterations of voltage-gated sodium channels, but also might implicate perturbation of PTX-sensitive Gαi/o-COupled signaling Wansduction pathways.展开更多
基金Acknowledgments This work was supported by a grant from The National Natural Science Foundation of China （30270884）. We greatly thank Dr Lai-Hua Xie （University of California at Los Angeles） for critical reading of the early draft of the manuscript. We are grateful to Dr Chang-Hui Rui （Institute of Plant Protection, CAAS） for technical assistance and suggestions.
文摘Pertussis toxin （FIX） inhibits the activation of the α-subunit of the inhibitory heterotrimeric G-proteins （Cαi/o） and modulates voltage-gated sodium channels, which may be one of the primary targets of pyrethroids. To investigate the potential mechanisms of agricultural pests resistance to pyrethroid insecticides, we examined the modulations by PTX on sodium channels in the central neurons of the 3rd-4th instar larvae of cyhalothrin-resistant （Cy-R） and cyhaiothrin-susceptible （Cy-S） Helicoverpa armigera by the whole-cell patch-clamp technique. The isolated neurons were cultured for 12-16 h in an improved L15 insect culture medium with or without PTX （400 ng/mL）. The results showed that both the Cy-R and Cy-S sodium channels exhibited fast kinetics and tetrodotoxin （TTX） sensitivity. The Cy-R sodium channels exhibited not only altered gating properties, including a 8.88-mV right shift in voltage-dependent activation （V0.5act） and a 6.54-mV right shift in voltage-dependent inactivation （V0.5inact）, but also a reduced peak in sodium channel density （Ⅰdensity） （55.2% of that in Cy-S neurons）. Cy-R sodium channels also showed low excitability, as evidenced by right shift of activation potential （Ⅴacti） by 5-10 mV and peak potential （Ⅴpcak） by 20 mV. FIX exerted significant effects on Cy-S sodium channels, reducing sodium channel density by 70.04%, right shifting V0.5act by 14.41 mV and V0.5inact by 9. 38 mV. It did not cause any significant changes of the parameters mentioned above in the Cy-R sodium channels. The activation time （Tpeak） from latency to peak at peak voltage and the fast inactivation time constant （τinact） in both Cy-S and Cy-R neurons were not affected. The results suggest that cotton bollworm resistant to pyrethroid insecticides involves not only mutations and allosteric alterations of voltage-gated sodium channels, but also might implicate perturbation of PTX-sensitive Gαi/o-COupled signaling Wansduction pathways.
