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First published online August 9, 2007
Journal of Experimental Biology 210, 2829-2835 (2007)
Published by The Company of Biologists 2007
doi: 10.1242/jeb.006064

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The visual pigments of a deep-sea teleost, the pearl eye Scopelarchus analis

Marie A. Pointer, Livia S. Carvalho, Jill A. Cowing, James K. Bowmaker and David M. Hunt^*

UCL Institute of Ophthalmology, 11–43 Bath Street, London, EC1V 9EL, UK

View larger version (44K): [in this window] [in a new window]   Fig. 1. Scopelarchus analis. (A) Photograph of the tubular eye, courtesy of Prof S. Collin. (B) Diagram of a vertical section through the eye. AR, accessory retina; L, lens; LP, lens pad; MR, main retina; ON, optic nerve. Redrawn from Partridge et al. (Partridge et al., 1992).

View larger version (34K): [in this window] [in a new window]   Fig. 2. Identity of RH1 opsins. (A) Alignment of RH1A and RH1B amino acid sequences. The deleted residues in RH1B that are involved in phosphorylation are indicated by arrowheads. (B) Alignment of 5' upstream region of RH1A and RH1B. The putative TATA boxes and start codons are underlined. (C) Phylogenetic tree of RH1 opsin nucleotide sequences generated by the neighbour-joining method. The RH2 sequences from Carassius auratus and extra-retinal sequences from Takifugu rubripes and Salmo salar were used as outgroups. All sequences were aligned with ClustalV. GenBank accession numbers for the sequences (in order from the top of the tree) are as follows: AJ249203, AJ249202, U12328, Y18677, L11863, S74449, AF105152, EF517407, EF517408, AY141256, AF201470, Y18666, AB185235, AB180742, NM_001078631, EF517404, EF517405, L11865, AF201469, NM_001033849.

View larger version (29K): [in this window] [in a new window]   Fig. 3. Identification of RH2 opsin. (A) Amino acid sequence of S. analis RH2 opsin aligned with the orthologous sequence from Metriaclima zebra. Note the additional Ser residues near the C terminus of the S. analis sequence. (B) Phylogenetic tree of RH2 opsin nucleotide sequences generated by the neighbour-joining method. The RH1 sequence from Carassius auratus was used as an outgroup. All sequences were aligned with ClustalV. Accession numbers for the sequences (in order from the top of the tree) are as follows: DQ088651, DQ088650, AY296739, Y18680, AY631037, AY771352, NM_001033712, AF38524, EF517406, AY599603, DQ088652, AB223053, AY214132, AF425076, AB098703, AB098704, AB087805, AB087806, AB087807, AB087808, L11865, AB110603, L11866, AB110602, L11863.

View larger version (41K): [in this window] [in a new window]   Fig. 4. Differential expression of RH1A and RH1B opsins. (A) Agarose gel electrophoresis separation of PCR products using RH1A- or RH1B-specific primers and cDNA from 300 m and 950 m caught fish. Note presence of RH1A transcripts in fish caught at both depths but RH1B transcripts only in fish caught at 950 m. (B) Relative levels of expression of RH1A and RH1B in two fish caught at 300 m and one at 950 m. Agarose gel electrophoresis separation of PCR products amplified using forward and reverse primers to identical sequences in RH1A and RH1B sequences. Amplified fragments are arrowed. Note smaller digested SalI fragments identifying RH1B transcripts only in cDNA from 950 m fish.

View larger version (17K): [in this window] [in a new window]   Fig. 5. Spectra of in vitro expressed RH1A and RH1B pigments. In both cases, a difference spectrum obtained by subtracting the bleached spectrum from the dark spectrum is shown. max values were obtained by fitting a visual pigment template to the difference spectrum (Govardovskii et al., 2000).