Abstract

Scallop retinas contain ciliary photoreceptor cells that respond to light by hyperpolarization like vertebrate rods and cones, but the response is generated by a different phototransduction cascade from those of rods and cones. To elucidate the cascade, we investigated a visual pigment and a G-protein functioning in the hyperpolarizing cell. Sequencing of cDNAs and in situ hybridization experiments showed that the hyperpolarizing cells express a novel subtype of visual pigment, which showed significant differences in amino acid sequence from other visual pigments. Cloning cDNA genes of G-protein and immunohistochemical analysis revealed the presence of an alpha subunit of a Go type G-protein, 83% identical in amino acid sequence to mammalian Go(alpha) in the nervous system, in the photoreceptive region of the cells. The results demonstrate that a novel, Go-mediated, phototransduction cascade is present in the hyperpolarizing cells. The phototransduction cascade in the scallop hyperpolarizing cell provides an alternative system to investigate Go-mediated transduction pathways in the nervous system. Molecular phylogenetic analysis strongly suggests that the Go-mediated phototransduction system emerged before the divergence of animals into vertebrate and invertebrate in the course of evolution.

Highlights

  • The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AB006454, AB006455, AB006456, and AB006457

  • One is the Gt1-mediated system of vertebrate hyperpolarizing photoreceptor cells in which the visual pigment activates a cGMP-specific phosphodiesterase via a heterotrimeric G-protein, transducin (Gt) [1,2,3]

  • After the first electrophysiological recordings of Hartline [10], the mechanism of the hyperpolarizing response as well as its evolution have been discussed in comparison with vertebrate hyperpolarizing ciliary photoreceptor cells, rods and cones [11,12,13,14]

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Summary

A Novel Go-mediated Phototransduction Cascade in Scallop Visual Cells*

Daisuke Kojima‡§¶, Akihisa Terakita‡, Toru Ishikawa‡, Yasuo Tsukaharaʈ, Akio Maeda‡, and Yoshinori Shichida‡**. Scallop retinas contain ciliary photoreceptor cells that respond to light by hyperpolarization like vertebrate rods and cones, but the response is generated by a different phototransduction cascade from those of rods and cones. In the photoreceptor cells of animals’ eyes, visual pigments trigger a G-protein-mediated phototransduction cascade, which eventually generates an electrical response of the cells. After the first electrophysiological recordings of Hartline [10], the mechanism of the hyperpolarizing response as well as its evolution have been discussed in comparison with vertebrate hyperpolarizing ciliary photoreceptor cells, rods and cones [11,12,13,14] It has been reported, that the hyperpolarizing response in the scallop cell is due to opening of a cGMPsensitive potassium channel [11,12,13,14], which is different from that of the vertebrate cells (closing of a cGMP-sensitive cationic channel) [15]. We show evidence that the phototransduction system in the invertebrate hyperpolarizing photoreceptor cells is not mediated by Gt or Gq, but rather uses a novel, Go-mediated cascade

EXPERIMENTAL PROCEDURES
A Novel Go-mediated Phototransduction
RESULTS AND DISCUSSION
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