Differential expression of the Enhancer of split genes in the developing Drosophila midgut.

The Notch signaling pathway plays an important role during development in animals from worms to humans and pathway components are required for the differentiation of many different cell types. In Drosophila, Su(H) dependent Notch activation up-regulates transcription of the Enhancer of split-Complex (E(spl)-C). The E(spl) genes are known to function during neurogenesis, although expression and genetics studies suggest that they also play roles in the development of other tissues. The majority of the E(spl) genes contain upstream binding sites for Su(H), proneural proteins, and E(spl) bHLH proteins resulting in overlapping expression patterns during embryonic development. However, their expression patterns are quite distinct during later embryonic stages and in larval imaginal discs. In order to characterize expression patterns of the E(spl) genes during development and determine potential mechanisms through which expression is controlled, we examined the expression levels and patterns of the E(spl) genes in the midgut during metamorphosis. Quantitative Reverse Transcriptase-PCR and X-Gal staining results show that the genes have different levels and patterns of expression in the developing midgut. Two ancestral E(spl) genes, malpha and mbeta, are highly expressed and increase significantly at puparium formation, whereas another gene, mgamma, is expressed at low levels and decreases in expression at puparium formation. We also show that mbeta is expressed in cells throughout the midgut, while mgamma is expressed in two small regions. These results provide further evidence that the E(spl) genes function during midgut development and that they are regulated by different factors.