Fig. 6. Novel, developmentally regulated expression of MsNSF. Pharate adult brains were analyzed by staining with antibodies against MsNSF (A,C,F), diuretic hormone (DH) (B,C), eclosion hormone (EH) (D,E) or bombyxin (A). Bombyxin is synthesized in four pairs of cells in the anterior protocerebrum (A, green) that do not colocalize with enriched MsNSF immunoreactivity (A, red). Diuretic hormone (DH) in later stages is localized to 80-100 neurosecretory cells (B), which lie medial to the eNSF-IR cells, as revealed by costaining with DH and MsNSF antibodies (C, arrows). EH in larvae is synthesized by paired ventro-medial cells (D). In later stages of development, EH-containing ventromedial cells in the brain migrate anteriorly, to the proctocerebrum, and are positioned ventrally to the eNSF-IR cells (E). Axons of eNSF-IR run parallel to EH axons to end in the CC—CA complex (F). The posterior aspect of brains stained for EH (E) or MsNSF is shown (F). Note the axons of eNSF-IR from Type IIa₄ cells run ventrally from the dorsally situated cell bodies (outside the plane of focus, arrow) in F; asterisks denote Type I staining.