Glial cells are one of major components of central nervous system. In the Drosophila central brain, each of five different subtypes of glial cells form specific organization called glial assembly. In the nervous system of holometabolous insects including Drosophila, neural networks remodels during metamorphosis. Then what would happen to the glial assembly during the neuronal remodelling? Here we report that the remodeling of two subtypes of neuropil-associated glia (ensheathing glia and astrocyte-like glia) during metamorphosis. Both neuropil of larval and adult brains are wrapped by ensheathing glia and filled with astrocyte-like glia. We found immature glial population appears in the interface between neuropil and cortex in the late larval stage. They proliferate and migrate in early pupal stage, while larval neuropil-associated glia changed their molecular and morphological features. When these immature glial cells were genetically labeled in larval stage, labels were specifically found in both subtypes of neuropil-associated glia in the adult brain. These results suggest that the larval and adult assemblies of neuropile-associated glia are composed of different cell population. Thus, the neuropil-associated glia assembly remodels during metamorphosis. We will discuss the interaction between mature larval glia and immature glia during the remodeling of glial assembly.