Mathematical analysis of phototransduction reaction parameters in rods and cones.

Retinal photoreceptor cells, rods and cones, convert photons of light into chemical and electrical signals as the first step of the visual transduction cascade. Although the chemical processes in the phototransduction system are very similar to each other in these photoreceptors, the light sensitivity and time resolution of the photoresponse in rods are functionally different than those in the photoresponses of cones. To systematically investigate how photoresponses are divergently regulated in rods and cones, we have developed a detailed mathematical model on the basis of the Hamer model.

Comparison of Bovine and Carp Fish Visual Pigment Photo-Intermediates at Room Temperature.

This paper presents room temperature nanoseconds to milliseconds time-resolved spectra and kinetics of the intermediate states and species of bovine and carp fish rhodopsin visual pigments, which also contained ~5% cone pigments. The nanoseconds to milliseconds range cover all the major intermediates in the visual phototransduction process except the formation of bathorhodopsin intermediate which occurs at the femtosecond time scale. The dynamics of these visual pigment intermediates are initiated by excitation with a 532 nm nanosecond laser pulse.

Molecular bases of rod and cone differences.

In the vertebrate retina, rods and cones both detect light, but they differ in functional aspects such as light sensitivity and temporal resolution, and in some cell biological aspects. For functional aspects, both types of photoreceptors use a phototransduction cascade, consisting of a series of enzymatic reactions, to convert photon capture to an electrical signal. To understand the mechanisms underlying the functional differences between rods and cones at the molecular level, we compared biochemically, each of the reactions in the phototransduction cascades of rods and cones using the cells isolated and purified from carp retina.

Transducin activates cGMP phosphodiesterase by trapping inhibitory γ subunit freed reversibly from the catalytic subunit in solution.

Activation of cGMP phosphodiesterase (PDE) by activated transducin α subunit (Tα*) is a necessary step to generate a light response in vertebrate photoreceptors. PDE in rods is a heterotetramer composed of two catalytic subunits, PDEα and PDEβ, and two inhibitory PDEγ subunits, each binding to PDEα or PDEβ. Activation of PDE is achieved by relief of the inhibitory constraint of PDEγ on the catalytic subunit.

Purification of cone outer segment for proteomic analysis on its membrane proteins in carp retina.

Rods and cones are both photoreceptors in the retina, but they are different in many aspects including the light response characteristics and, for example, cell morphology and metabolism. These differences would be caused by differences in proteins expressed in rods and cones. To understand the molecular bases of these differences between rods and cones, one of the ways is to compare proteins expressed in rods and cones, and to find those expressed specifically or dominantly.

Dephosphorylation during bleach and regeneration of visual pigment in carp rod and cone membranes.

On absorption of light by vertebrate visual pigment, the chromophore, 11-cis retinal, is isomerized to all-trans retinal to activate the phototransduction cascade, which leads to a hyperpolarizing light response. Activated pigment is inactivated by phosphorylation on the protein moiety, opsin. Isomerized all-trans retinal is ultimately released from opsin, and the pigment is regenerated by binding to 11-cis retinal.

Phosphorylation-independent suppression of light-activated visual pigment by arrestin in carp rods and cones.

Visual pigment in photoreceptors is activated by light. Activated visual pigment (R*) is believed to be inactivated by phosphorylation of R* with subsequent binding of arrestin. There are two types of photoreceptors, rods and cones, in the vertebrate retina, and they express different subtypes of arrestin, rod and cone type.

RDH13L, an enzyme responsible for the aldehyde-alcohol redox coupling reaction (AL-OL coupling reaction) to supply 11-cis retinal in the carp cone retinoid cycle.

Cone photoreceptors require effective pigment regeneration mechanisms to maintain their sensitivity in the light. Our previous studies in carp cones suggested the presence of an unconventional and very effective mechanism to produce 11-cis retinal, the necessary component in pigment regeneration. In this reaction (aldehyde-alcohol redox coupling reaction, AL-OL coupling reaction), formation of 11-cis retinal, i.

Quantitative aspects of cGMP phosphodiesterase activation in carp rods and cones.

Cones are less light-sensitive than rods. We showed previously in carp that more light (>100-fold) is required in cones than in rods to activate 50% of cGMP phosphodiesterase (PDE). The lower effectiveness of PDE activation in carp cones is due partly to the fact that the activation rate of transducin (Tr) by light-activated visual pigment (R*) is 5-fold lower in carp cones than in rods.

Substrate specificity and subcellular localization of the aldehyde-alcohol redox-coupling reaction in carp cones.

Our previous study suggested the presence of a novel cone-specific redox reaction that generates 11-cis-retinal from 11-cis-retinol in the carp retina. This reaction is unique in that 1) both 11-cis-retinol and all-trans-retinal were required to produce 11-cis-retinal; 2) together with 11-cis-retinal, all-trans-retinol was produced at a 1:1 ratio; and 3) the addition of enzyme cofactors such as NADP(H) was not necessary. This reaction is probably part of the reactions in a cone-specific retinoid cycle required for cone visual pigment regeneration with the use of 11-cis-retinol supplied from Müller cells.

Different phosphorylation rates among vertebrate cone visual pigments with different spectral sensitivities.

Cone photoreceptor subtypes having different spectral sensitivities exhibit different recovery kinetics in their photoresponses in some vertebrates. Phosphorylation by G protein-coupled receptor kinase (GRK) is essential for the rapid inactivation of light-activated visual pigment, which is the rate-limiting step of the cone photoresponse recovery in salamander. In this study we compared the rate of light-dependent phosphorylation by GRK7 of carp green- and blue-sensitive cone visual pigments.

Larger inhibition of visual pigment kinase in cones than in rods.

In the carp retina, visual pigment kinase, GRK1 (G-protein coupled receptor kinase 1) in rods and GRK7 in cones, is inhibited by a photoreceptor neuronal Ca(2+)-sensor protein, S-modulin (or recoverin) in rods and visinin (formerly named s26) in cones. Here, we compared Ca(2+)-dependent inhibition of GRK1 by S-modulin and that of GRK7 by visinin. First, the concentrations of S-modulin and visinin in the outer segment were estimated: the concentration of visinin (1.

High cGMP synthetic activity in carp cones.

Cones show briefer light responses than rods and do not saturate even under very bright light. Using purified rod and cone homogenates, we measured the activity of guanylate cyclase (GC), an enzyme responsible for cGMP synthesis and therefore recovery of a light response. The basal GC activity was 36 times higher in cones than in rods: It was mainly caused by higher expression levels of GC in cones (GC-C) than in rods (GC-R).

Highly efficient retinal metabolism in cones.

After bleaching of visual pigment in vertebrate photoreceptors, all-trans retinal is reduced to all-trans retinol by retinol dehydrogenases (RDHs). We investigated this reaction in purified carp rods and cones, and we found that the reducing activity toward all-trans retinal in the outer segment (OS) of cones is >30 times higher than that of rods. The high activity of RDHs was attributed to high content of RDH8 in cones.

Rod and cone photoreceptors: molecular basis of the difference in their physiology.

Vertebrate retinal photoreceptors consist of two types of cells, the rods and cones. Rods are highly light-sensitive but their flash response time course is slow, so that they can detect a single photon in the dark but are not good at detecting an object moving quickly. Cones are less light-sensitive and their flash response time course is fast, so that cones mediate daylight vision and are more suitable to detect a moving object than rods.

Identification of differentially expressed genes in carp rods and cones.

Purpose: Rods and cones differ in their photoresponse characteristics, morphology, and susceptibilities to certain diseases. To contribute to the studies at the molecular level of these differences, we tried to identify genes expressed preferentially in rods or cones.

Methods: From purified carp rods and cones, we extracted their RNA and obtained corresponding cDNA pools (rod cDNA and cone cDNA).

Amino acid residues in GRK1/GRK7 responsible for interaction with S-modulin/recoverin.

GRK1 is a visual pigment kinase in rods and is essential for inactivation of light-activated rhodopsin. The GRK1 activity is inhibited by binding of the Ca(2+)-bound form of S-modulin/recoverin. We previously identified the S-modulin/recoverin site to interact with GRK1.

Molecular mechanisms characterizing cone photoresponses.

In the vertebrate retina, rods mediate twilight vision and cones mediate daylight vision. Their photoresponse characteristics are different. The light-sensitivity of a cone is 10(2)-10(3) times lower than that of a rod.

Highly effective phosphorylation by G protein-coupled receptor kinase 7 of light-activated visual pigment in cones.

Cone photoreceptors show briefer photoresponses than rod photoreceptors. Our previous study showed that visual pigment phosphorylation, a quenching mechanism of light-activated visual pigment, is much more rapid in cones than in rods. Here, we measured the early time course of this rapid phosphorylation with good time resolution and directly compared it with the photoresponse time course in cones.

Neuronal calcium sensor proteins are direct targets of the insulinotropic agent repaglinide.

The NCS (neuronal calcium sensor) proteins, including neurocalcins, recoverins and visinin-like proteins are members of a family of Ca2+-sensitive regulators, each with three Ca2+-binding EF-hand motifs. In plants, lily CCaMK [chimaeric Ca2+/CaM (calmodulin)-dependent protein kinase] and its PpCaMK ( Physcomitrella patens CCaMK) homologue are characterized by a visinin-like domain with three EF-hands. In the present study, in an effort to discover NCS antagonists, we screened a total of 43 compounds using Ca2+-dependent drug affinity chromatography and found that the insulinotropic agent repaglinide targets the NCS protein family.

Stimulatory effect of cyanidin 3-glycosides on the regeneration of rhodopsin.

Anthocyanins have been suggested to improve visual functions. This study examined the effect of four anthocyanins in black currant fruits on the regeneration of rhodopsin using frog rod outer segment (ROS) membranes. Cyanidin 3-glycosides, glucoside and rutinoside, stimulated the regeneration, but the corresponding delphinidins showed no significant effect.

S-modulin.

S-Modulin is a Ca2+-binding protein found in frog rod photoreceptors (1,2) and its bovine homologue is known as recoverin (3,4). In the Ca2+-bound form, S-modulin inhibits rhodopsin phosphorylation5 through inhibition of rhodopsin kinase. (6-9) Because rhodopsin phosphorylation is the quench mechanism of light-activated rhodopsin (R*), (10,11) the inhibition of the phosphorylation by S-modulin probably contributes to increase the lifetime of R* to result in sustained hydrolysis of cGMP5.