Molecular and functional identification of a novel photopigment in Pecten ciliary photoreceptors

• Published in: The Journal of general physiology Vol. 150; no. 3; pp. 401 - 415
• Main Authors: Arenas, Oscar, Osorno, Tomás, Malagón, Gerardo, Pulido, Camila, Gomez, María Del Pilar, Nasi, Enrico
• Format: Journal Article
• Language: English
• Published: United States Rockefeller University Press 05-03-2018
• Subjects: Complementary DNA; Cyclic nucleotides; Gene expression; Hybridization; Nucleotides; Opsins; Photopigments; Photoreceptors; Pigments; Polymerase chain reaction; Retina; Rhodopsin; RNA-mediated interference; Signal transduction; siRNA; Transcription; Visual perception; Western blotting
• ISSN: 0022-1295; 1540-7748
• DOI: 10.1085/jgp.201711938

The two basic animal photoreceptor types, ciliary and microvillar, use different light-transduction schemes: their photopigments couple to G
versus G
proteins, respectively, to either mobilize cyclic nucleotides or trigger a lipid signaling cascade. A third class of photoreceptors has been described in the dual retina of some marine invertebrates; these present a ciliary morphology but operate via radically divergent mechanisms, prompting the suggestion that they comprise a novel lineage of light sensors. In one of these organisms, an uncommon putative opsin was uncovered that was proposed to signal through G
Orthologues subsequently emerged in diverse phyla, including mollusks, echinoderms, and chordates, but the cells in which they express have not been identified, and no studies corroborated their function as visual pigments or their suggested signaling mode. Conversely, in only one invertebrate species,
, have the ciliary photoreceptors been physiologically characterized, but their photopigment has not been identified molecularly. We used the transcriptome of
retina to guide the cloning by polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE) extensions of a new member of this group of putative opsins. In situ hybridization shows selective transcription in the distal retina, and specific antibodies identify a single band of the expected molecular mass in Western blots and distinctly label ciliary photoreceptors in retina sections. RNA interference knockdown resulted in a reduction in the early receptor current-the first manifestation of light transduction-and prevented the prolonged aftercurrent, which requires a large buildup of activated rhodopsin. We also obtained a full-length clone of the α-subunit of a G
from
retina complementary DNA and localized it by in situ hybridization to the distal photoreceptors. Small interfering RNA targeting this G
caused a specific depression of the photocurrent. These results establish this novel putative opsin as a bona fide visual pigment that couples to G
to convey the light signal.