To facilitate the investigation of glial development inDrosophila, we present a detailed description of theDrosophila glial cells in the ventral nerve cord. A GAL4 enhancer-trap screen for glial-specific expression was performed. Using UAS-lacZ and UAS-kinesin-lacZ as reporter constructs, we describe the distribution and morphology of the identified glial cells in the fully differentiated ventral nerve cord of first-instar larvae just after hatching. The three-dimensional structure of the glial network was reconstructed using a computer. Using the strains with consistent GAL4 expression during late embryogenesis, we traced back the development of the identified cells to provide a glial map at embryonic stage 16. We identify typically 60 (54-64) glial cells per abdominal neuromere both in embryos and early larvae. They are divided into six subtypes under three categories: surface-associated glia (16-18 subperineurial glial cells and 6-8 channel glial cells), cortex-associated glia (6-8 cell body glial cells), and neuropile-associated glia (8-10 nerve root glial cells, 14-16 interface glial cells, and 3-4 midline glial cells). The proposed glial classification system is discussed in comparison with previous insect glial classifications.