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Fig. 2. SSCs express tph1. (A) Expression pattern of tph1 mRNA at stage 31. Signals were detected in the brain and in a punctate pattern in the epidermis. A sagittal histological section (A′) demonstrated tph1expression in the epiphysis (A′) in SSCs and in a subset of neuronal cells (n) in the floor plate of the neural tube (A′′). (B) SSCs express tph1 (blue arrowhead), as demonstrated by whole-mount in situ hybridization followed by IF for serotonin (green, green arrowhead) and cilia (acetylated-α-tubulin, red, red arrowhead). (C) Punctate expression pattern of aromatic-L-amino-acid decarboxylase (DOPA-decarboxylase; ddc) in the embryonic skin. Inset shows close-up. Embryos are shown in lateral views. a, anterior; d, dorsal; epi, epiphysis; l, left; r, right. |
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Fig. 3. Inhibition of serotonin synthesis impairs cilia-driven flow. (A) Experimental setup. Embryos were incubated with TPH inhibitor p-chloro-phenylalanin (PCPA) from stage 25-32, followed by assessment of flow (B), serotonin localization (C) and tph1 expression (D). (B) Reduction of flow velocity in PCPA-treated specimen was partially rescued by co-culture with exogenously applied serotonin. Flow was analyzed by data processing from time-lapse movies following the addition of fluorescent beads to the medium. The mean velocity in control embryo was set to 100%. (C) Absence of serotonin staining (red arrowheads) upon PCPA incubation (C′). (D) Unaltered tph1 expression following PCPA treatment (D′). |
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Fig. S2. Embryonic expression of serotonin pathway components.
(A, B) Epidermal expression of monoamine transporter slc18a1 mRNA in stage 14 neurula (A) and stage 35/36 tadpole (B) embryos. Transverse sections (A', B') reveal uniform superficial staining in the epidermis. Note lack of expression in the neural plate at stage 14. (C) tph2 expression was restricted to the hindbrain at stage 30. (C') Transverse histological section. Embryos are shown in lateral views except for panel (A), which shows a dorsal perspective. (D, D') tph1 mRNA expressing SCCs (blue arrowheads) secrete PNA positive vesicles (red arrowheads). (D) Close-up bright field image of a transverse section of a tph1 stained tadpole at stage 36 displaying three SSCs. (D') Fluorescent PNA signals (red) co-localize with tph1 mRNA. |
