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First published online April 18, 2008
Journal of Experimental Biology 211, 1495-1503 (2008)
Published by The Company of Biologists 2008
doi: 10.1242/jeb.012047

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The influence of ontogeny and light environment on the expression of visual pigment opsins in the retina of the black bream, Acanthopagrus butcheri

Julia Shand^1,*, Wayne L. Davies², Nicole Thomas¹, Lois Balmer¹, Jill A. Cowing², Marie Pointer², Livia S. Carvalho², Ann E. O. Trezise³, Shaun P. Collin⁴, Lyn D. Beazley¹ and David M. Hunt²

¹ School of Animal Biology, University of Western Australia, Crawley, WA 6009, Australia
² UCL Institute of Ophthalmology, 11–43 Bath Street, London EC1V 9EL, UK
³ Australian Equine Genetics Research Centre, School of Biomedical Sciences, University of Queensland, Brisbane, Qld 4072, Australia
⁴ Sensory Neurobiology Group, School of Biomedical Sciences, University of Queensland, Brisbane, Qld 4072, Australia

View larger version (14K): [in this window] [in a new window]   Fig. 1. Spectral characteristics of the lighting used for the two different rearing conditions. (A) Light intensity with broad spectrum fluorescent lights, plus a neutral density filter (0.2 optical density) around the tanks to equate the intensity of light with that of the short wavelength-reduced conditions (shown in C). (B) The transmission characteristics of the yellow filter used to reduce short wavelength light. (C) Light intensity with the tanks surrounded by the yellow filter (*note the reduction in the peaks at the shorter wavelengths).

View larger version (12K): [in this window] [in a new window]   Fig. 2. Changes in the frequency of cones during development in fish reared in standard lighting conditions. The individual max values from all cones scanned, as determined by curve fitting, are plotted at 1 nm intervals.

View larger version (21K): [in this window] [in a new window]   Fig. 3. Phylogenetic tree for visual opsin gene sequences. The tree was generated by the neighbour-joining method (Saitou and Nei, 1987) using amino acid sequences aligned by ClustalW (Higgins et al., 1996). The degree of support for internal branching was assessed by bootstrapping with 1000 replicates using the MEGA2 computer package (Kumar et al., 2001). GenBank accession numbers for the sequences (from top to bottom) are EU090913, EU090914, AY296739, DQ235683, DQ088651, DQ235682, DQ088650, AB223053, AB223054, AB223055, DQ0088652, DQ235681, AY214132, AB098703, AB098704, L11865, L11866, AF201470, L11863, DQ354577, AB180742, AY296738, AY296736, AB223057, AF247118, DQ354581, AY214134, L11864, DQ354580, AB223056, AF247114, AY296737, AY214131, L11867, DQ354578, AB223051, AY296739, D85863, AY214133, DQ354579, AY296735, AB223058, NM_057353.

View larger version (9K): [in this window] [in a new window]   Fig. 4. Quantification of opsin gene transcript levels in the larval and adult retina. Each bar shows the mean (±s.e.m.) relative expression of each opsin in either the larva or adult. The values were obtained from separate RNA preparations from four adult retinae and three pools of 20 larval heads. For each opsin, the difference in relative expression between larva and adult was statistically significant (Student's t-test) at the 1% probability level.

View larger version (12K): [in this window] [in a new window]   Fig. 5. Comparison of the frequency of cone classes measured by microspectrophotometry and opsin gene expression relative to total cone opsin expression in the larval and adult retina.

View larger version (6K): [in this window] [in a new window]   Fig. 6. Absorption spectra for Rh2A and Rh2Aβ pigments generated in vitro. The difference spectrum (dark minus bleached) for each pigment is shown as a fitted Govardovskii template (Govardovskii et al., 2000). The max values for Rh2A and Rh2Aβ are 527 and 534 nm, respectively.

View larger version (27K): [in this window] [in a new window]   Fig. 7. Frequency of cone classes at different developmental stages. Data from all four stages for fish reared under standard conditions are shown. No data were available for wild-caught larval fish or for adult yellow filter fish.