Shifting roles ofDrosophilapair-rule gene orthologs: segmental expression and function in the milkweed bugOncopeltus fasciatus

The discovery of pair-rule genes (PRGs) in Drosophila revealed the existence of an underlying two-segment-wide prepattern directing embryogenesis. The milkweed bug Oncopeltus , a hemimetabolous insect, is a more representative arthropod: most of its segments form sequentially after gastrulation. Here we report the expression and function of orthologs of the complete set of nine Drosophila PRGs in Oncopeltus . Seven Of -PRG-orthologs are expressed in stripes in the primordia of every segment, rather than every-other segment, Of-runt is PR-like, and several are also expressed in the segment addition zone. RNAi-mediated knockdown of Of-odd-skipped , paired and sloppy-paired impacted all segments, with no indication of PR-like register. We confirm that Of - E75A is expressed in PR-like stripes, although it is not PR in Drosophila , demonstrating the existence of an underlying PR-like prepattern in Oncopeltus . These findings reveal that a switch occurred in regulatory circuits leading to segment formation: while several holometabolous insects are “ Drosophila -like,” utilizing PRG-orthologs for PR-patterning, most Of- PRGs are expressed segmentally in Oncopeltus , a more basally-branching insect. Thus, an evolutionarily stable phenotype – segment formation – is directed by alternate regulatory pathways in diverse species. Summary Statement Despite the broad of conservation of segmentation in insects, the regulatory genes underlying this process in Drosophila have different roles in the hemipteran, Oncopeltus fasciatus .