This study investigates the reactivation of neural stem cells (NSCs) in the Drosophila brain in response to hormonal stimuli. By dissecting freshly hatched larvae and culturing their brains, researchers can analyze NSC proliferation and the effects of specific hormones.
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.
Dissect the brains out of the larvae placed in the second glass watch dish with SSM, using forceps under a dissecting microscope and adjusting the magnification as needed. Use one forceps to grab the mouth hooks, and with the other, gently grab the body halfway down, and pull in the opposite direction to split the larva into two pieces. After dissecting 15 to 20 brains, add 1 milliliter of SSM into one well of a sterile 24-well culture tray. Using a micropipette and a sterile tip, transfer the freshly dissected brains into the SSM, followed by incubation of the media with brains for 24 hours at 25 degrees Celsius.
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