Figure 3

Identification of peptidergic neurons and their projections in the larval CNS. (a) Strong expression of AST-CC in cells 27/704 of the TG2 detected by ISH. (b, b´) Low levels of AST-CC transcript in IN-704 neurons of AG3 identified with antibodies to CCAP and MIPs (arrows; yellow). (c, c’) Lateral VL8 neurons in the TAG (arrowheads; yellow) identified by ISH with sNPF probe followed by double staining with antibodies to MIP (red) and RFamide (green). VL8 axons project via ventral nerves (small arrows; yellow) and arborize on muscle surface of 9th segment (d). Numerous varicosities in axon terminals indicate neuropeptide release from these putative neurohemal sites. (e) Medial PM8 and PM9 neurons detected by ISH with CCH1 probe (arrows). (f) Antibody staining confirmed colocalization of CCH1 (red) and CT (green) in PM8 and PM9 neurons (arrows; yellow) that project axons via proctodeal nerves (arrowheads) and (g) innervate the hindgut. These medial neurons are also stained with MIP antibody (c’; large arrows; red). Scale bars a-c´, e, f = 50 µm, d = 25 µm, g = 300 µm.