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Fig. 4 Ectopic gene expression in the pineal gland driven by the zebrafish rh chimeric promoter carrying the PIPE sequence. (A) Schematic representation of the constructs, Rh(–1084), Rh(–1084)/PIPE, and PIPE-Rh(–1084). (B) Comparison of PIPE and nearby sequence in Ex(–1055) with those in the corresponding region of Rh(–1084) or Rh(–1084)/PIPE. Bold characters in Rh(–1084)/PIPE represent five nucleotides (four substitutions and a single insertion) modified from Rh(–1084) to create an ectopic PIPE sequence. (C–F) Dorsal views (anterior up) of Rh(–1084) (C and D) and Rh(–1084)/PIPE (E and F) transgenic larvae at 7 dpf. Nomarski (C and E) and fluorescent (D and F) images were taken at the same focal plane without moving the larvae. EGFP fluorescence signals in the pineal gland of Rh(–1084)/PIPE transgenic larva are marked by an arrowhead (F). (F Inset) High-magnification image of the EGFP-positive pineal structure. The larvae in C–F were not treated with 1-phenyl-2-thiourea, so that strong EGFP fluorescence signals in the pigmented eyes (D and F) are only visible through the pupil. [Bars = 100 μm (C–F), 20 μm (F Inset).]