Figure 4. Whole-mount IF of Platynereis larvae with antibodies raised against neuropeptides labels distinct subsets of neuronal tracks in the VNC.
(A) Ventral view of Platynereis VNC, stained with an anti-acetylated tubulin antibody. This is the registered average VNC generated from scans of 36 larvae. (B–L) Whole-mount IF of Platynereis larvae with an antibody raised against (B) FVa, (C) RGWa, (D) FVRIa, (E) RYa, (F) PDF, (G) LEUC, (H) LUQ, (I) MIP, (J) ATO, (K) PENK, (L) FMRFa. Whole-mount scans were cropped to show only the VNC region. Scale bar: 20 μm.
Figure 4—figure supplement 1. Morpholino-mediated knockdown of proneuropeptides followed by whole-mount IF indicates antibody specificities.
Whole-mount IF of 72 hpf uninjected control Platynereis larvae and larvae micro-injected with translation-blocking morpholinos (MOs) targeting different neuropeptide precursor genes. FVa-knockdown and control larvae were stained with an antibody raised against FVa and counterstained with acetylated tubulin. All other control and knockdown larvae were co-stained with an antibody raised against FVa and an antibody raised against the MO target peptide indicated and counterstained with acetylated tubulin. All larvae are shown in ventral view. For each neuropeptide, three control larvae (top) and three MO-knockdown larvae (bottom) are shown. (A) FVa knockdown and control larvae with antibody raised against FVa. (B) FMRFa knockdown and control larvae with antibody raised against FMRFa. (C) LUQ knockdown and control larvae with antibody raised against LUQ. (D) ATO knockdown and control larvae with antibody raised against ATO. (E) FVRIa knockdown and control larvae with antibody raised against FVRIa. (F) PDF knockdown and control larvae with antibody raised against PDF. (G) RGWa knockdown and control larvae with antibody raised against RGWa. Note that the staining in the head is eliminated following knockdown. The staining in the VNC was too weak in these samples. (H) RYa knockdown and control larvae with antibody raised against RYa. (I) PENK knockdown and control larvae with antibody raised against PENK. (J) LEUC knockdown and control larvae with antibody raised against LEUC. Scale bar: 30 µm.