| Summary: | Glia are crucial for the normal development and functioning of the nervous system in many animals. Insects are widely used for studies of glia genetics and physiology. <i>Drosophila melanogaster</i> surface glia (perineurial and subperineurial) form a blood–brain barrier in the central nervous system and blood–nerve barrier in the peripheral nervous system. Under the subperineurial glia layer, in the cortical region of the central nervous system, cortex glia encapsulate neuronal cell bodies, whilst in the peripheral nervous system, wrapping glia ensheath axons of peripheral nerves. Here, we show that the expression of the evolutionarily conserved <i>swiss cheese</i> gene is important in several types of glia. <i>swiss cheese</i> knockdown in subperineurial glia leads to morphological abnormalities of these cells. We found that the number of subperineurial glia nuclei is reduced under <i>swiss cheese</i> knockdown, possibly due to apoptosis. In addition, the downregulation of <i>swiss cheese</i> in wrapping glia causes a loss of its integrity. We reveal transcriptome changes under <i>swiss cheese</i> knockdown in subperineurial glia and in cortex + wrapping glia and show that the downregulation of <i>swiss cheese</i> in these types of glia provokes reactive oxygen species acceleration. These results are accompanied by a decline in animal mobility measured by the negative geotaxis performance assay.
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