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Stop the crop: insights into the insecticidal mode of action of cinnamodial against mosquitoes

By Megha Kalsi, Anton Walter, Beenhwa Lee, Andrew DeLaat, Renata Rusconi Trigueros, Katharina Happel, Rose Sepesy, Bao Nguyen, Preston K. Manwill, H. Liva Rakotondraibe, Peter M. Piermarini

Posted 31 May 2020
bioRxiv DOI: 10.1101/2020.05.30.125203 (published DOI: 10.1016/j.pestbp.2020.104743)

Cinnamodial (CDIAL) is a drimane sesquiterpene dialdehyde found in the bark of Malagasy medicinal plants ( Cinnamosma species; family Canellaceae). We previously demonstrated that CDIAL was insecticidal, antifeedant, and repellent against Aedes aegypti mosquitoes. The goal of the present study was to generate insights into the insecticidal mode of action for CDIAL, which is presently unknown. We evaluated the effects of CDIAL in vitro on the contractility of the ventral diverticulum (crop) in adult female Ae. aegypti . The crop is a food storage organ surrounded by visceral muscle that spontaneously contracts in vitro . We found that CDIAL completely inhibited spontaneous contractions of the crop as well as those stimulated by the agonist 5-hydroxytryptamine. Several derivatives of CDIAL with known insecticidal activity also inhibited crop contractions. Morphometric analyses of crops suggested that CDIAL induced a tetanic paralysis that was dependent on extracellular Ca2+ and inhibited by Gd3+, a non-specific blocker of plasma membrane Ca2+ channels. Screening of numerous pharmacological agents revealed that a Ca2+ ionophore (A23187) was the only compound other than CDIAL to completely inhibit crop contractions via a tetanic paralysis. Taken together, our results suggest that CDIAL inhibits crop contractility by elevating intracellular Ca2+ through the activation of plasma membrane Ca2+ channels thereby leading to a tetanic paralysis, which may explain the insecticidal effects of CDIAL against mosquitoes. Our pharmacological screening efforts also revealed the presence of two regulatory pathways in mosquito crop contractility not previously described: an inhibitory glutamatergic pathway and a stimulatory octopaminergic pathway. The latter was also completely inhibited by CDIAL. ### Competing Interest Statement The authors have declared no competing interest.

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