To explore astrocytic ALKBH5 modulate depression-related behaviors via glutamatergic neuronal activity, we bidirectionally injected the AAV-CamkIIα-hM4D(Gi)-mCherry virus into the mPFC of Astrocyte cKO mice to specifically manipulate the activity of glutamatergic neurons. This evidence concerns the gene ALKBH5 and major depressive disorder.