Effects of Guadipyr on voltage-gated calcium and potassium channels in central neurons of Helicoverpa armigera

doi:10.7679/j.issn.2095-1353.2020.086

Abstract

Abstract:

[Objectives] Guadipyr is a novel insecticide that has both neonicotinoid and semicarbazone insecticidal activity. How it affects ion channels and channel gating characteristics and functions is currently unknown. We examined the effects of Guadipyr on voltage-gated calcium and potassium channels, and explored whether these channels are potential targets for this pesticide. [Methods] The effects of three concentrations (1, 10, and 100 μmol?L ^–1) of Guadipyr on Ca ²⁺and K ⁺ channels were studied using the patch clamp technique. [Results] All three concentrations of Guadipyr significantly (P<0.05) shifted Ca ²⁺channel Ⅰ-Ⅴ and activation curves about 10-15 mV in the negative direction. The inactivation curves of Ca ²⁺channels were also shifted 5 mV towards the hyperpolarizing direction, which is a not significant shift (P>0.05). The Ca ²⁺peak current was significantly decreased by Guadipyr treatment relative to the control. The I_peak decreased more slowly as the concentration of Guadipyr increased. The area of window current increased the most (93.20%) following treatment with 1 μmol?L ^–1 Guadipyr than by treatment with 10 μmol?L ^–1 and 100 μmol?L ^–1. The number of Ca ²⁺ channels activated increased after Guadipyr treatment at a test voltage of ﹣60 mV to +10 mV. The K ⁺peak current decreased gradually when Guadipyr was added to the external solution; the I_peak decreased more slowly as the Guadipyr concentration increased from 10 μmol?L ^–1 to 100 μmol?L ^–1. Ⅰ-Ⅴ curves declined and the activation of K ⁺channels was shifted 8 mV towards the depolarizing direction, which was not significant (P>0.05). This shows that K ⁺ channels can be activated at higher potentials by Guadipyr. [Conclusion] Guadipyr can effectively inhibit the I_peak of the Ca ²⁺and K ⁺channels, and shift the activation and inactivation curves of these channels, thereby affecting the gating of Ca ²⁺ channels and K ⁺ channels. Therefore, Ca ²⁺ and K ⁺ channels in the central neurons of H. armigera are potential Guadipyr target sites.

Key words: Guadipyr, Helicoverpa armigera, voltage-gated calcium channels, voltage-gated potassium channels, Whole-cell Patch Clamp Technique

Dan-Yang GUAN, Xiao-Wei JIANG, Qing-Ya LI, Xiao LIU, Yong-Qiang MA, Qiang CHEN, Hongmei LI-Byarlay, Bing-Jun HE. Effects of Guadipyr on voltage-gated calcium and potassium channels in central neurons of Helicoverpa armigera[J]. Chinese Journal of Applied Entomology, 2020, 57(4): 841-849.

Figures/Tables 11

Fig. 1 ICa of the Helicoverpa armigera before and after the application of Gua A. Control; B. ICa after Gua (10 μmol?L–1).

Fig. 2 TheⅠ-Ⅴcurves and Ipeak of Ca2+ channels before and after the application of Gua A. Gua's effects on the Ⅰ-Ⅴ curve of Ca2+ channels: Standardized current amplitude; B. The Ipeak of Ca2+ channels after Gua. * indicats significant difference compared with the control (P<0.05, One-way analysis of variance). The same as Fig.7.

Fig. 3 Effects of Gua on the steady-state activation of ICa

Table 1 Effects of Gua on the steady-state activation of ICa

条件 Condition	激活 Activation
条件 Condition	V_0.5^a	k^b	n
对照 Control	–9.83±0.73	4.55±0.38	10
1 μmol?L^–1戊吡虫胍 1 μmol?L^–1Gua	–22.85±5.02*	5.27±0.26	5
10 μmol?L^–1戊吡虫胍 10 μmol?L^–1Gua	–23.45±3.36*	5.48±0.21	8
100 μmol?L^–1戊吡虫胍 100 μmol?L^–1Gua	–24.21±3.74*	5.46±0.26	5

Fig. 4 Effects of Gua on the steady-state inactivation of ICa

Table 2 Effects of Gua on the steady-state inactivation of ICa

条件 Condition	失活 Inactivation
条件 Condition	V_0.5^a	k^b	n
对照 Control	–22.30±2.46	18.45±3.60	7
1 μmol?L^–1戊吡虫胍 1 μmol?L^–1Gua	–26.26±2.86	13.58±3.36	5
10 μmol?L^–1戊吡虫胍 10 μmol?L^–1Gua	–24.76±2.44	12.28±2.95	6
100 μmol?L^–1戊吡虫胍 100 μmol?L^–1Gua	–28.69±3.12	13.41±3.13	5

Fig. 5 Comparative analysis about the effect of Gua on the Window current of ICa A. 1 μmol?L–1 Gua; B. 10 μmol?L–1 Gua; C. 100 μmol?L–1 Gua.

Fig. 6 IK of the Helicoverpa armigera before and after the application of Gua A. Control; B. IK after Gua (100 μmol?L–1).

Fig. 7 TheⅠ-Ⅴ curves and Ipeak of IK before and after the application of Gua A. Gua’s effects on the I-V curve of K+ channels: Current amplitude is not standardized; B. The Ipeak of K+ channels after Gua.

Fig. 8 Effects of Gua on the steady-state activation of IK

Table 3 Effects of Gua on the steady-state activation of IK

条件 Condition	激活 Activation
条件 Condition	V_0.5^a	k^b	n
对照Control	22.15±1.72	21.10±0.74	13
10 μmol?L^–1戊吡虫胍 10 μmol?L^–1Gua	31.72±1.07*	22.31±0.78	7
100 μmol?L^–1戊吡虫胍 100 μmol?L^–1Gua	29.35±1.55	24.16±1.93	4

References 24

[1]	Ahmad M, Gladwell RT, McCaffery AR , 1989. Decreased nerve sensitivity is a mechanism of resistance in a pyrethroid resistant strain of Heliothis armigera from Thailand. Pesticide Biochemistry and Physiology, 35(2):165-171.
[2]	Ahmad M, McCaffery AR , 1991. Elucidation of detoxication mechansims involved in resistance to insecticides in the third instar larvae of field-selected strain of Helicoverpa armigera with the use of synergists. Pesticide Biochemistry and Physiology, 41(1):41-52.
[3]	Bloomquist JR , 1996. Ion channels as targets for insecticides. Annual Review of Entomology, 41:163-190. doi: 10.1146/annurev.en.41.010196.001115 URL pmid: 8546445
[4]	Cao Z, Shafer TJ, Murray TF , 2010. Mechanisms of pyrethroid insecticide-induced stimulation of calcium influx in neocortical neurons. Journal of Pharmacology and Experimental Therapeutics, 336(1):197-205. doi: 10.1124/jpet.110.171850 URL pmid: 20881019
[5]	Clark JM, Symington SB , 2007. Pyrethroid action on calcium channels: Neurotoxicological implications. Invertebrate Neuroscience, 7(1):3-6. URL pmid: 17294162
[6]	Hamill OF, Marty A, Neher E, Sakmann B, Sigworth FJ , 1981. Improved patch clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Archiv European Journal of Physiology, 391(2):85-100. URL pmid: 6270629
[7]	He BJ, Chen JT, Guo SY, Rui HH, Meng XQ, Wang XL, Wang YN, Sun JS, Liu AX , 2001. Nerve cell culture and ultrastrucal analysis of central nerve genglia from resistant Helicoverpa armigera. Acta Scientiarum Naturalium Universitatis Nankaiensis, 34(1):30-36.
	[ 贺秉军, 陈家童, 郭世宜, 芮黄辉, 孟香清, 王秀玲, 王亦农, 孙金生, 刘安西 , 2001. 抗性棉铃虫神经细胞的离体培养和超微结构分析. 南开大学学报, 34(1):30-36.]
[8]	He BJ, Liu AX, Chen JT, Sun JS, Rui CH , 2002. Study on the mechanism of action of cyhalothrin on sodium and calcium channels of nerve cells from Helicoverpa armigera. Acta Biophysica Sinica, 18(2):201-205.
	[ 贺秉军, 刘安西, 陈家童, 孙金生, 芮昌辉 , 2002. 三氟氯氰菊酯对棉铃虫神经细胞钠及钙通道作用机理研究. 生物物理学报, 18(2):201-205.]
[9]	He XW, Yin RY, Chen YH, LV J, Xie ZP, He FS , 1997. Effect of pyrethroids on Na⁺、Ca²⁺ channel currents in rat brain neurons. Chinese Journal of Industrial Hygiene and Occupational Diseases, 15(5):261-264.
	[ 贺锡文, 殷若元, 陈寅红, 吕京, 谢佐平, 何凤生 , 1997. 拟除虫菊酯对神经细胞膜Na⁺、Ca²⁺离子通道的影响. 中华劳动卫生职业病杂志, 15(5):261-264.]
[10]	Li DZ, Wang K, Xu L, Chai TT, Cui F, Qiu LH, Zheng MQ, Qin ZH, Wang CJ , 2006. Acute toxicity of guadipyr to non-target organisms. Asian Journal of Ecotoxicology, 11(3):331-337.
	[ 李冬植, 王凯, 徐莉, 柴婷婷, 崔峰, 邱立红, 郑明奇, 覃兆海, 王成菊 , 2006. 戊吡虫胍对几种非靶标生物的急性毒性. 生态毒理学报, 11(3):331-337.]
[11]	Liu AX, Chen ST , 1990. Modified action of cypermethrin enantiomers on axonal sodium and potassium channels of Periplaneta Fulginosa(Serville). Acta Entomologica Sinica, 33(1):1-6.
	[ 刘安西, 陈守同 , 1990. 氯氰菊酯异构体对黑胸大蠊神经钠钾通道的调制作用. 昆虫学报, 33(1):1-6.]
[12]	Neal AP, Yuan Y, Atchison WD , 2010. Allethrin differentially modulates voltage-gated calcium channel subtypes in rat PC12 cells. Toxicological Sciences, 116(2):604-613. doi: 10.1093/toxsci/kfq139 URL pmid: 20466778
[13]	Nishimura K, Kanda Y, Okazawa A, Ueno T , 1994. Relationship between insecticidal and neyrophysiological activities of imidacloprid and related compounds. Pesticide Biochemistry and Physiology, 50(1):51-59. doi: 10.1006/pest.1994.1057 URL
[14]	Qi SZ, Wang C, Chen XF, Qin ZH, Li XF, Wang CJ , 2013. Toxicity assessments with Daphnia magna of Guadipyr, a new neonicotinoid insecticide and studies of its effect on acetylcholinesterase (AChE), glutathione-S-transferase (GST), catalase (CAT) and chitobiase activities. Ecotoxicology and Environmental Safety, doi.org/ 10.1016/j.ecoenv.2013.09.013. doi: 10.1016/j.ecoenv.2020.111192 URL pmid: 32858326
[15]	Rao GV, Rao KS , 1997. Modulation of K⁺ transport across synaptosomes of rat brain by synthetic pyrethroids . Journal of the Neurological Sciences, 147(2):127-133. doi: 10.1016/s0022-510x(96)05327-0 URL pmid: 9106117
[16]	Salgado VL , 1992. The neurotoxic insecticidal mechanism of the nonsterodial ecdysone agonist RH-5849: K⁺ channel block in nerve and muscle. Pestic Biochem. Physiol., 43(1):1-13. doi: 10.1016/0048-3575(92)90013-P URL
[17]	Shang XL , 2014. Effects of tefluthrin and deltamethrin on calcium channels and BK channels in central neurons isolated from Helicoverpa armigera. Master dissertation. Tianjin: Nankai University.
	[ 商学良 , 2014. 七氟菊酯和溴氰菊酯对棉铃虫神经细胞Ca ²+通道及Ca ²+激活K +通道(BK)作用机理的研究 . 硕士学位论文. 天津: 南开大学.]
[18]	Sheng CF, Xuan WJ, Su JW, Wang HT , 2001. A protracted disaster of cotton bollworm in China and the role of sex pheromone in the disaster reduction. Journal of Natural Disasters, 10(1):75-79.
	[ 盛承发, 宣维健, 苏建伟, 王红托 , 2001. 棉铃虫灾害的长期性及性信息素的减灾控害作用. 自然灾害学报, 10(1):75-79.]
[19]	Soderlund DM, Clark JM, Sheets L P, Mullin LS, Piccirillo VJ, Sargent D, Stevens JT, Weiner ML , 2001. Mechanisms of pyrethroid neurotoxicity: Implications for cumulative risk assessment. Toxicology, 171(1):3-59. doi: 10.1016/s0300-483x(01)00569-8 URL pmid: 11812616
[20]	Su W, Zhou Y, Ma Y, Wang L, Zhang Z, Rui C, Duan H, Qin Z , 2012. N'-Nitro-2-hydrocarbyliden-ehydrazinecarboximidamides: Design, synthesis, crystal structure, insecticidal activity, and structure- activity relationships. Journal of Agricultural & Food Chemistry, 60(20):5028-5034. doi: 10.1021/jf300616x URL pmid: 22546079
[21]	Von Stein RT, Silver KS, Soderlund DM , 2013. Indoxacarb, metaflumizone, and other sodium channel inhibitor insecticides: Mechanism and site of action on mammalian voltage-gated sodium channels. Pesticide Biochemistry and Physiology, 106(3):101-112. doi: 10.1016/j.pestbp.2013.03.004 URL pmid: 24072940
[22]	Wang Y, He BJ, Zhao Q, Liang Z, Liu AX , 2006a. Effects of cyhalothrin on the transient outward potassium current in central neurons of Helicoverpa armigera. Insect Science, 13(1):13-17. doi: 10.1111/ins.2006.13.issue-1 URL
[23]	Wang Y, He BJ, Wu CL, Liu AX , 2006b. Inhibition effects of cyhaliothrin on the delayed rectifier potassium current in the central neurons of Helicoverpa armigera. Acta Entomologica Sinica, 49(2):235-240.
[24]	Zhang MF, Fan JY, Zhang HW, Zhang XZ, Ma XG , 2009. Research development of studies on neonicotinoid insecticides. World Pesticides, 31(1):34-37, 64.
	[ 张梅凤, 范金勇, 张宏伟, 张秀珍, 马新刚 , 2009. 新烟碱类杀虫剂的研究进展. 世界农药, 31(1):34-37, 64.]