Abstract
FlyNap (triethylamine) is commonly used to anesthetize Drosophila melanogaster fruit flies. The purpose of this study was to determine whether triethylamine is a suitable anesthetic agent for research into circulatory physiology and immune competence in the mosquito, Anopheles gambiae (Diptera: Culicidae). Recovery experiments showed that mosquitoes awaken from traditional cold anesthesia in less than 7 minutes, but that recovery from FlyNap anesthesia does not begin for several hours. Relative to cold anesthesia, moderate exposures to FlyNap induce an increase in the heart rate, a decrease in the percentage of the time the heart contracts in the anterograde direction, and a decrease in the frequency of heartbeat directional reversals. Experiments employing various combinations of cold and FlyNap anesthesia then showed that cold exposure does not affect basal heart physiology, and that the differences seen between the cold and the FlyNap groups are due to a FlyNap-induced alteration of heart physiology. Furthermore, exposure to FlyNap eliminated the cardioacceleratory effect of crustacean cardioactive peptide (CCAP), and reduced a mosquito’s ability to survive a bacterial infection. Together, these data show that FlyNap is not a suitable substitute to cold anesthesia in experiments assessing mosquito heart function or immune competence. Moreover, these data also illustrate the intricate biology of the insect heart. Specifically, they confirm that the neurohormone CCAP modulates heart rhythms and that it serves as an anterograde pacemaker.
Highlights
A common need in experiments evaluating biological processes in mosquitoes is the complete immobilization of the insect
We found that exposure to FlyNap moderately elevates heart rates, biases the proportional directionality of heart contractions, eliminates the heart’s response to the cardioacceleratory neuropeptide crustacean cardioactive peptide (CCAP), and decreases the ability of mosquitoes to survive the early stages of a bacterial infection
5% of the mosquitoes recovered by 1 min after being returned to room temperature, 60% recovered by 4 min, and 100% recovered by 7 min (Figure 1A)
Summary
A common need in experiments evaluating biological processes in mosquitoes is the complete immobilization of the insect. The gold standard for mosquito immobilization is cold-induced anesthesia [1,2,3], mosquitoes are frequently anesthetized by exposure to CO2 [4,5]. Both of these methods, while effective, have their limitations. The purpose of this study was to determine whether FlyNap (triethylamine) is a suitable anesthetic agent for mosquito research
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