Abstract
Laboratories that study Drosophila melanogaster or other insects commonly use carbon dioxide (CO2) anaesthesia for sorting or other work. Unfortunately, the use of CO2 has potential unwanted physiological effects, including altered respiratory and muscle physiology, which impact motor function behaviours. The effects of CO2 at different levels and exposure times were examined on the subsequent recovery of motor function as assessed by climbing and flight assays. With as little as a five minute exposure to 100% CO2, D. melanogaster exhibited climbing deficits up to 24 hours after exposure. Any exposure length over five minutes produced climbing deficits that lasted for days. Flight behaviour was also impaired following CO2 exposure. Overall, there was a positive correlation between CO2 exposure length and recovery time for both behaviours. Furthermore, exposure to as little as 65% CO2 affected the motor capability of D. melanogaster. These negative effects are due to both a CO2-specific mechanism and an anoxic effect. These results indicate a heretofore unconsidered impact of CO2 anaesthesia on subsequent behavioural tests revealing the importance of monitoring and accounting for CO2 exposure when performing physiological or behavioural studies in insects.
Highlights
Laboratories that study Drosophila melanogaster or other insects commonly use carbon dioxide (CO2) anaesthesia for sorting or other work
For appropriate selection of an anaesthetic method to use in insect behavioural and physiological studies, it is important to determine if anoxia and CO2 anaesthesia have separate effects and how persistent these effects are
The results of the study reveal: 1) there is a chronic negative effect of a short exposure to 100% CO2 on climbing (Fig 1a), which is potentially indefinite in length; 2) there is a long-lasting effect of 100% CO2 exposure on flight for CO2 exposures of 15 minutes or longer (Fig. 2); and 3) the mechanism behind these detrimental effects is a combination of a CO2-specific mechanism and an anoxic effect (Figs 1 and 3)
Summary
Laboratories that study Drosophila melanogaster or other insects commonly use carbon dioxide (CO2) anaesthesia for sorting or other work. For appropriate selection of an anaesthetic method to use in insect behavioural and physiological studies, it is important to determine if anoxia and CO2 anaesthesia have separate effects and how persistent these effects are. This study determined the effects of CO2, hypoxia and anoxia on D. melanogaster climbing and flight behaviours, which are routinely used to assay motor function and performance in flies.
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