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Fig. 5
Ectopic Hcrt neurons project to the locus coeruleus. (A) Confocal projection of a 120hpf Tg(hcrt:Kaede, dbh:EGFP) larva injected with an hs:lhx9 plasmid shows that ectopic photoconverted Hcrt cells in the medial hindbrain (red) project to the locus coeruleus (LC). (B) Confocal projection of a 120hpf Tg(dbh:EGFP) larva injected with an elavl3:Kaede plasmid to stochastically label neurons shows that some neurons in the medial hindbrain project to the LC (arrowhead) whereas others do not (arrow; axon projects orthogonally to the image). (C) All ectopic Hcrt neurons project to the LC, but only 20% of elavl3:Kaede-labeled cells in the same brain region project to the LC. n, number of neurons analyzed. ***P<0.001 compared with elavl3:Kaede (Fisher′s exact test). Yellow boxes in A,B indicate the regions used for quantification. (D) Schematic of hcrt, pdyn and vglut1 expression in the ventral zebrafish brain at 120 hpf. The locations of ectopic Hcrt neurons, dbh-expressing LC neurons and the photoconverted region are shown. Scale bars: 10µm.