Drosophila Burrowing and Tunneling Assay: een methode om weefselhypoxie bij vlieglarven te beoordelen

- To set up the embryo collection cage, use grape juice agar plates supplemented with fresh yeast paste and place the plates onto cages containing male and female Drosophila flies of the appropriate genotype. The odor from the yeast and grape juice is attractive and promotes egg laying by the females. Allow egg laying for a timed period. Then remove the adults and incubate the embryo plates for 24 hours to obtain first instar larvae. Then gently transfer a few individual larvae from the grape juice plate onto an agar-only test plate where a hole in the agar in the center of the plate was cut out and filled with yeast paste.

Young larvae must feed to gain body weight to develop, so they burrow in the food source, the yeast paste. As they develop, larvae enter a wandering phase and tunnel away from the food to seek out a site for pupation. To assess hypoxia, examine the tunneling patterns larvae form in the substrate. Lack or insufficient tunneling is a sign of oxygen deprivation. In the example protocol, we will see how to set up the burrowing and tunneling assay.

- Set up the control and experimental genetic crosses in egg lay dishes as described in the text protocol. Keep the flie