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First published online January 25, 2005
Journal of Experimental Biology 208, 433-438 (2005)
Published by The Company of Biologists 2005
doi: 10.1242/jeb.01420

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Pigmentation in the sensory organs of the ascidian larva is essential for normal behavior

Di Jiang^1,*, Jason W. Tresser^1,*, Takeo Horie², Motoyuki Tsuda² and William C. Smith¹

¹ Department of Molecular, Cellular and Developmental Biology, University of California, Santa Barbara, California 93106, USA
² Department of Life Science, Graduate School of Life Science, University of Hyogo, 3-2-1 Kouto, Kamigori, Ako-gun, Hyogo 678-1297, Japan

View larger version (68K): [in a new window]   Fig. 1. Larvae of the ascidian Ciona savignyi. (A) Wild type, (B) homozygous immaculate (C) homozygous spotless. oto, otolith; oce, ocellus. Scale bars, 50 µm.

View larger version (75K): [in a new window]   Fig. 2. Pigmentation in the sensory vesicle of Ciona savignyi imc and spt mutants is deficient. (A) Diagram of the Ciona sensory vesicle. (B,D,F) Nomarski images of the otolith in wild type (B), homozygous imc (D) and homozygous spt (F). (C,E,G) Nomarski images of the ocellus in wild type (C), homozygous imc (E) and homozygous spt (G). lc, lens cells; p, pigmented cell in ocellus; ph, photoreceptors; Scale bars, 20 µm.

View larger version (58K): [in a new window]   Fig. 3. Immunohistochemical staining of opsin1 (red) and arrestin (green) in wild type (A), homozygous imc (B) and homozygous spt (C) Swimming larva. Scale bars, 20 µm.

View larger version (53K): [in a new window]   Fig. 4. L-dopa staining of the sensory vesicle in wild type (A), homozygous imc (B), and homozygous spt (C) Swimming larva. Scale bars, 20 µm.

View larger version (73K): [in a new window]   Fig. 5. spt mutation and rescue. (A) Schematic diagram of Ciona savignyi tyrosinase protein structure and the site of mutation in spt. (B,C) Pigmentation in otolith (B) and ocellus (C) in a homozygous spt tadpole rescued by electroporation of wild-type C. savignyi tyrosinase genomic DNA. Scale bars, 10 µm.

View larger version (25K): [in a new window]   Fig. 6. Homozygous imc and spt tadpoles failed to swim towards shaded areas and against gravity. (A-C) Diagrams of the apparatus; (A) top view; (B,C) side views. (A,B) For phototaxis assays Petri dishes were half shaded (black) and larvae were placed in the shaded area and their response monitored. (C) For geotaxis assays tadpoles were placed in Petri dish containing 60 ml filtered seawater with antibiotics. A second Petri dish was suspended on top of the first dish at a height of 1 cm to sandwich the larvae between two potential settling surfaces. (D,E) Percentage of tadpoles settled on either dark (black) or illuminated (white) area. (F,G) Percentage of tadpoles settled on either top (black) or bottom (white) of the apparatus.