title: Cyclic nucleotide-gated ion channels in rod photoreceptors are protected from retinoid inhibition.
abstract: In vertebrate rods, photoisomerization of the 11-cis retinal chromophore of rhodopsin to the all-trans conformation initiates a biochemical cascade that closes cGMP-gated channels and hyperpolarizes the cell. All-trans retinal is reduced to retinol and then removed to the pigment epithelium. The pigment epithelium supplies fresh 11-cis retinal to regenerate rhodopsin. The recent discovery that tens of nanomolar retinal inhibits cloned cGMP-gated channels at low [cGMP] raised the question of whether retinoid traffic across the plasma membrane of the rod might participate in the signaling of light. Native channels in excised patches from rods were very sensitive to retinoid inhibition. Perfusion of intact rods with exogenous 9- or 11-cis retinal closed cGMP-gated channels but required higher than expected concentrations. Channels reopened after perfusing the rod with cellular retinoid binding protein II. PDE activity, flash response kinetics, and relative sensitivity were unchanged, ruling out pharmacological activation of the phototransduction cascade. Bleaching of rhodopsin to create all-trans retinal and retinol inside the rod did not produce any measurable channel inhibition. Exposure of a bleached rod to 9- or 11-cis retinal did not elicit channel inhibition during the period of rhodopsin regeneration. Microspectrophotometric measurements showed that exogenous 9- or 11-cis retinal rapidly cross the plasma membrane of bleached rods and regenerate their rhodopsin. Although dark-adapted rods could also take up large quantities of 9-cis retinal, which they converted to retinol, the time course was slow. Apparently cGMP-gated channels in intact rods are protected from the inhibitory effects of retinoids that cross the plasma membrane by a large-capacity buffer. Opsin, with its chromophore binding pocket occupied (rhodopsin) or vacant, may be an important component. Exceptionally high retinoid levels, e.g., associated with some retinal degenerations, could overcome the buffer, however, and impair sensitivity or delay the recovery after exposure to bright light.
authors.affiliation: Department of Ophthalmology, Massachusetts Eye and Ear Infirmary and Harvard Medical School, Boston, MA 02114, USA.
authors.forename: Quanhua
authors.initials: Q
authors.lastname: He
authors.forename: Dmitriy
authors.initials: D
authors.lastname: Alexeev
authors.forename: Maureen E
authors.initials: ME
authors.lastname: Estevez
authors.forename: Sarah L
authors.initials: SL
authors.lastname: McCabe
authors.forename: Peter D
authors.initials: PD
authors.lastname: Calvert
authors.forename: David E
authors.initials: DE
authors.lastname: Ong
authors.forename: M Carter
authors.initials: MC
authors.lastname: Cornwall
authors.forename: Anita L
authors.initials: AL
authors.lastname: Zimmerman
authors.forename: Clint L
authors.initials: CL
authors.lastname: Makino
mesh_terms: D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000378:metabolism; D000557:Ambystoma; D000818:Animals; D006152:Cyclic GMP / Q000096:biosynthesis; D054815:Cyclic Nucleotide-Gated Cation Channels; D004224:Diterpenes; D006162:Guanylate Cyclase / Q000378:metabolism; D007473:Ion Channels / Q000037:antagonists & inhibitors / Q000502:physiology*; D008027:Light; D015591:Microspectrophotometry; D018408:Patch-Clamp Techniques; D017948:Retinal Rod Photoreceptor Cells / Q000187:drug effects / Q000502:physiology* / Q000528:radiation effects; D012172:Retinaldehyde / Q000378:metabolism / Q000494:pharmacology; D012176:Retinoids / Q000378:metabolism / Q000494:pharmacology*; D012177:Retinol-Binding Proteins / Q000494:pharmacology; D054839:Retinol-Binding Proteins, Plasma; D012243:Rhodopsin / Q000378:metabolism; D012374:Rod Cell Outer Segment / Q000378:metabolism; D014801:Vitamin A / Q000494:pharmacology
chemical_list: D054815:Cyclic Nucleotide-Gated Cation Channels; D004224:Diterpenes; D007473:Ion Channels; D012176:Retinoids; D012177:Retinol-Binding Proteins; D054839:Retinol-Binding Proteins, Plasma; D014801:Vitamin A; C031390:9-cis-retinal; D012243:Rhodopsin; D015106:3',5'-Cyclic-GMP Phosphodiesterases; D006162:Guanylate Cyclase; D006152:Cyclic GMP; D012172:Retinaldehyde; D015056:1-Methyl-3-isobutylxanthine