Lokale hittesondetest: een methode om nociceptie van hoge temperaturen bij Drosophila-larven te beoordelen

- To prepare Drosophila larvae for a heat probe assay, collect animals of the desired developmental stage, and transfer single larvae onto a test plate with a thin film of water. The water allows the larvae to move freely without sticking to the plate surface.

Drosophila larvae detect noxious thermal stimuli through multidendritic sensory neurons in their epidermis known as nociceptors. These don't fire until a certain sensory threshold is reached, such as potentially harmful high temperatures.

Set the heat probe to a temperature that will overcome this sensory threshold, and cause the nociceptors to fire. Gently press the tip of the probe against the larva to apply heat. The larva will respond to the noxious temperature with thermo-nociceptive behaviors, withdrawing from the probe. Stop when the larva exhibits a 360-degree role away from the heat probe, or when a defined time has elapsed. Record the response time.

In the example protocol, we will see a demonstration of a localized heat probe assay on early third instar Drosophila larvae.

- First, preset the temperature of the thermal probe to the desired set point. Use a paintbrush or forceps to gently transfer an individual larva