pitx2c morphants display larger parapineals. (A,B) Single confocal sections of the pineal complex showing in situ hybridisation against gfi1.2 in control and pitx2c morphant embryos at 72 hpf. The outline of the pineal gland is highlighted with a dotted line. (C) Counts of cell expressing gfi1.2 in control and pitx2c morphants. The number of gfi1.2-positive cells is significantly increased in the morphant context; **P<0.01 using a t-test. Error bars represent s.e.m. (D-D22) Single confocal sections extracted from a time-lapse series showing the migrating parapineal (indicated with a dotted line) in a Tg(foxd3:GFP)zf15 embryo; nuclei are labelled in red with Histone2B:RFP. At 24 hpf (D), the parapineal has only just delaminated from the anterior region of the pineal complex. Leftward migration is clearly engaged at 6 hours (D2) and continues throughout the analysis (D2,D22). Time after the initiation of the analysis is indicated on each frame. (E-E22) Single confocal sections extracted from a time-lapse series showing the migrating parapineal (indicated with a dotted line) in a pitx2c morphant embryo carrying the Tg(foxd3:GFP)zf15 transgene; nuclei are labelled in red with Histone2B:RFP. As in wild-type siblings, pitx2c morphant parapineals have migrated to the left at 6 hours. Time after the initiation of the analysis is indicated on each frame. (F) Numbers of cells in the parapineal during migration in wild-type (green) and pitx2c morphant embryos (orange). A statistically significant difference in parapineal cell numbers is detected in the morphant versus the control as early as 2 hours into the time-lapse datasets. *P>0.1; ***P<0.0.001 using a two-way repeated measures ANOVA test. Error bars represent s.e.m. Embryos are viewed dorsally with the anterior towards the top.
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