Analyzing Neural Stem Cell Reactivation in Cultured Drosophila Brain Explants

In response to external stimuli, Drosophila brain neural stem cells, or NSCs, exit quiescence, a state of cell cycle arrest, and reactivate to begin proliferation.

Take freshly hatched Drosophila larvae, a stage where NSCs naturally exit quiescence in vivo.

Grasp the mouth hooks and the body to split the larva.

Locate the brain behind the mouth hooks and excise it.

Transfer the brains to culture plate wells containing media.

In treated wells, the media contains a test hormone, while control wells lack the hormone. Incubate.

Under control conditions, the NSCs remain quiescent due to the absence of external stimuli.

In treated wells, the hormone binds to specific NSC receptors, activating downstream signaling pathways and reactivating the NSCs.

The reactivated NSCs enter the cell cycle and undergo proliferation.

Post-incubation, the brains are now ready for downstream analysis to visualize NSC proliferation, confirming the hormone's reactivation potential.