| The developing midline of Drosophila consists of diverse cell types that must migrate and differentiate appropriately to form a functional central nervous system. Despite the paucity of midline cells, much is unknown about how these cells migrate to their final locations and acquire fates. In a previous RNA-sequencing screen, we found that 18-Wheeler (18w), a Toll-family receptor, is present in the midline during embryonic development. Fluorescence in situ hybridizations and genetic analyses revealed that 18w is expressed posteriorly in segments in the median neuroblast. 18w overexpression led to midline disorganization, while heterozygous 18w loss of function mutants showed no phenotype, indicating that one wild-type copy of 18w is sufficient to maintain function.;To further analyze midline gene expression, a protocol was optimized to dissociate cells from tissue for fluorescence-activated cell sorting. Isolated midline cells would subsequently undergo single-cell RNA-sequencing, revealing expression differences among cell types and allowing for transcriptome analysis. |
