RESUMEN
The retinal circadian system consists of a network of clocks located virtually in every retinal cell-type. Although it is established that the circadian clock regulates many rhythmic processes in the retina, the links between retinal cell-specific clocks and visual function remain to be elucidated. Bmal1 is a principal, non-redundant component of the circadian clock in mammals and is required to keep 24 h rhythms in the retinal transcriptome and in visual processing under photopic light condition. In the current study, we investigated the retinal function in mice with a rod-specific knockout of Bmal1. For this purpose, we measured whole retina PER2::Luciferase bioluminescence and the dark-adapted electroretinogram (ERG). We observed circadian day-night differences in ERG a- and b-waves in control mice carrying one allele of Bmal1 in rods, with higher amplitudes during the subjective night. These differences were abolished in rod-specific Bmal1 knockout mice, whose ERG light-responses remained constitutively low (day-like). Overall, PER2::Luciferase rhythmicity in whole retinas was not defective in these mice but was characterized by longer period and higher rhythmic power compared to retinas with wild type Bmal1 gene. Taken together, these data suggest that a circadian clock located in rods regulates visual processing in a cell autonomous manner.
Asunto(s)
Relojes Circadianos/fisiología , Adaptación a la Oscuridad/fisiología , Células Fotorreceptoras Retinianas Bastones/metabolismo , Factores de Transcripción ARNTL/genética , Animales , Electrorretinografía , Femenino , Regulación de la Expresión Génica/fisiología , Luciferasas/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Visión Nocturna/fisiología , Proteínas Circadianas Period/metabolismo , Estimulación Luminosa , Reacción en Cadena en Tiempo Real de la Polimerasa , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , Rodopsina/genética , Sinaptofisina/genéticaRESUMEN
Rod photoreceptors can be saturated by exposure to bright background light, so that no flash superimposed on the background can elicit a detectable response. This phenomenon, called increment saturation, was first demonstrated psychophysically by Aguilar and Stiles and has since been shown in many studies to occur in single rods. Recent experiments indicate, however, that rods may be able to avoid saturation under some conditions of illumination. We now show in ex vivo electroretinogram and single-cell recordings that in continuous and prolonged exposure even to very bright light, the rods of mice from both sexes recover as much as 15% of their dark current and that responses can persist for hours. In parallel to recovery of outer segment current is an â¼10-fold increase in the sensitivity of rod photoresponses. This recovery is decreased in transgenic mice with reduced light-dependent translocation of the G protein transducin. The reduction in outer-segment transducin together with a novel mechanism of visual-pigment regeneration within the rod itself enable rods to remain responsive over the whole of the physiological range of vision. In this way, rods are able to avoid an extended period of transduction channel closure, which is known to cause photoreceptor degeneration.SIGNIFICANCE STATEMENT Rods are initially saturated in bright light so that no flash superimposed on the background can elicit a detectable response. Frederiksen and colleagues show in whole retina and single-cell recordings that, if the background light is prolonged, rods slowly recover and can continue to produce significant responses over the entire physiological range of vision. Response recovery occurs by translocation of the G protein transducin from the rod outer to the inner segment, together with a novel mechanism of visual-pigment regeneration within the rod itself. Avoidance of saturation in bright light may be one of the principal mechanisms the retina uses to keep rod outer-segment channels from ever closing for too long a time, which is known to produce photoreceptor degeneration.
Asunto(s)
Células Fotorreceptoras Retinianas Bastones/metabolismo , Transducina/metabolismo , Animales , Electrorretinografía , Femenino , Luz , Masculino , Ratones , Transporte de Proteínas , Células Fotorreceptoras Retinianas Bastones/fisiología , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , Análisis de la Célula Individual , Transducina/genética , Visión OcularRESUMEN
Photobiomodulation (PBM) by far-red (FR) to near-infrared (NIR) light has been demonstrated to restore the function of damaged mitochondria, increase the production of cytoprotective factors and prevent cell death. Our laboratory has shown that FR PBM improves functional and structural outcomes in animal models of retinal injury and retinal degenerative disease. The current study tested the hypothesis that a brief course of NIR (830 nm) PBM would preserve mitochondrial metabolic state and attenuate photoreceptor loss in a model of retinitis pigmentosa, the P23H transgenic rat. P23H rat pups were treated with 830 nm light (180 s; 25 mW/cm2; 4.5 J/cm2) using a light-emitting diode array (Quantum Devices, Barneveld, WI) from postnatal day (p) 10 to p25. Sham-treated rats were restrained, but not treated with 830 nm light. Retinal metabolic state, function and morphology were assessed at p30 by measurement of mitochondrial redox (NADH/FAD) state by 3D optical cryo-imaging, electroretinography (ERG), spectral-domain optical coherence tomography (SD-OCT), and histomorphometry. PBM preserved retinal metabolic state, retinal function, and retinal morphology in PBM-treated animals compared to the sham-treated group. PBM protected against the disruption of the oxidation state of the mitochondrial respiratory chain observed in sham-treated animals. Scotopic ERG responses over a range of flash intensities were significantly greater in PBM-treated rats compared to sham controls. SD-OCT studies and histological assessment showed that PBM preserved the structural integrity of the retina. These findings demonstrate for the first time a direct effect of NIR PBM on retinal mitochondrial redox status in a well-established model of retinal disease. They show that chronic proteotoxic stress disrupts retinal bioenergetics resulting in mitochondrial dysfunction, and retinal degeneration and that therapies normalizing mitochondrial metabolism have considerable potential for the treatment of retinal degenerative disease.
Asunto(s)
Metabolismo Energético/efectos de la radiación , Terapia por Luz de Baja Intensidad/métodos , Mitocondrias/efectos de la radiación , Degeneración Retiniana/radioterapia , Retinitis Pigmentosa/radioterapia , Animales , Modelos Animales de Enfermedad , Electrorretinografía , Flavina-Adenina Dinucleótido/metabolismo , Rayos Infrarrojos , Mitocondrias/metabolismo , NAD/metabolismo , Oxidación-Reducción , Ratas , Ratas Transgénicas , Degeneración Retiniana/diagnóstico por imagen , Degeneración Retiniana/metabolismo , Degeneración Retiniana/patología , Células Fotorreceptoras Retinianas Bastones/metabolismo , Células Fotorreceptoras Retinianas Bastones/patología , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , Retinitis Pigmentosa/diagnóstico por imagen , Retinitis Pigmentosa/metabolismo , Retinitis Pigmentosa/patología , Tomografía de Coherencia Óptica , Resultado del TratamientoRESUMEN
Noninvasive, objective measurement of rod function is as significant as that of cone function, and for retinal diseases such as retinitis pigmentosa and age-related macular degeneration, rod function may be a more sensitive biomarker of disease progression and efficacy of treatment than cone function. Functional imaging of single human rod photoreceptors, however, has proven difficult because their small size and rapid functional response pose challenges for the resolution and speed of the imaging system. Here, we describe light-evoked, functional responses of human rods and cones, measured noninvasively using a synchronized adaptive optics optical coherence tomography (OCT) and scanning light ophthalmoscopy (SLO) system. The higher lateral resolution of the SLO images made it possible to confirm the identity of rods in the corresponding OCT volumes.
Asunto(s)
Luz , Oftalmoscopía/métodos , Células Fotorreceptoras Retinianas Conos/citología , Células Fotorreceptoras Retinianas Conos/efectos de la radiación , Células Fotorreceptoras Retinianas Bastones/citología , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , HumanosRESUMEN
Photoreceptors are specialized cells devoted to the transduction of the incoming visual signals. Rods are able also to shed from their tip old disks and to synthesize at the base of the outer segment (OS) new disks. By combining electrophysiology, optical tweezers (OTs), and biochemistry, we investigate mechanosensitivity in the rods of Xenopus laevis, and we show that 1) mechanosensitive channels (MSCs), transient receptor potential canonical 1 (TRPC1), and Piezo1 are present in rod inner segments (ISs); 2) mechanical stimulation-of the order of 10 pN-applied briefly to either the OS or IS evokes calcium transients; 3) inhibition of MSCs decreases the duration of photoresponses to bright flashes; 4) bright flashes of light induce a rapid shortening of the OS; and 5) the genes encoding the TRPC family have an ancient association with the genes encoding families of protein involved in phototransduction. These results suggest that MSCs play an integral role in rods' phototransduction.
Asunto(s)
Fototransducción , Mecanotransducción Celular , Células Fotorreceptoras Retinianas Bastones/metabolismo , Xenopus laevis/metabolismo , Animales , Calcio/metabolismo , Fluorescencia , Luz , Fototransducción/efectos de la radiación , Mecanotransducción Celular/efectos de la radiación , Familia de Multigenes , Estimulación Luminosa , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , Canales Catiónicos TRPC/genética , Proteínas de Xenopus/genéticaRESUMEN
Retinitis pigmentosa (RP) is a debilitating blinding disease affecting over 1.5 million people worldwide, but the mechanisms underlying this disease are not well understood. One of the common models used to study RP is the retinal degeneration-10 (rd10) mouse, which has a mutation in Phosphodiesterase-6b (Pde6b) that causes a phenotype mimicking the human disease. In rd10 mice, photoreceptor cell death occurs with exposure to normal light conditions, but as demonstrated in this study, rearing these mice in dark preserves their retinal function. We found that inactivating rhodopsin signaling protected photoreceptors from degeneration suggesting that the pathway activated by this G-protein-coupled receptor is causing light-induced photoreceptor cell death in rd10 mice. However, inhibition of transducin signaling did not prevent the loss of photoreceptors in rd10 mice reared under normal light conditions implying that the degeneration caused by rhodopsin signaling is not mediated through its canonical G-protein transducin. Inexplicably, loss of transducin in rd10 mice also led to photoreceptor cell death in darkness. Furthermore, we found that the rd10 mutation in Pde6b led to a reduction in the assembled PDE6αßγ2 complex, which was corroborated by our data showing mislocalization of the γ subunit. Based on our findings and previous studies, we propose a model where light activates a non-canonical pathway mediated by rhodopsin but independent of transducin that sensitizes cyclic nucleotide gated channels to cGMP and causes photoreceptor cell death. These results generate exciting possibilities for treatment of RP patients without affecting their vision or the canonical phototransduction cascade.
Asunto(s)
Muerte Celular , Luz , Degeneración Retiniana/patología , Células Fotorreceptoras Retinianas Bastones/patología , Rodopsina/metabolismo , Transducina/fisiología , cis-trans-Isomerasas/fisiología , Animales , GMP Cíclico/metabolismo , Modelos Animales de Enfermedad , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Degeneración Retiniana/genética , Degeneración Retiniana/metabolismo , Células Fotorreceptoras Retinianas Bastones/metabolismo , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , Retinitis Pigmentosa , Transducción de SeñalRESUMEN
Retinal ganglion cells (GCs) are important visual neurons which carry complex spatiotemporal information from the retina to higher visual centers in the brain. By taking advantage of pathway-specific knockout/mutant mice and multi-electrode array (MEA) recording techniques, we analyze contributions of rod and cone pathways to responsiveness, kinetics and receptive field profiles of GCs under scotopic and photopic conditions. Our data suggest: (1) Scotopic responses of some GCs require all three rod pathways, some require only the secondary and tertiary rod pathways, and others require only the tertiary rod pathway. (2) There are more responsive GCs in photopic conditions than responsive GCs in scotopic conditions. (3) Gap junctions slow down GCs' scotopic light responses and increase GCs' ratio of antagonistic to center inputs. (4) Cone pathways do not affect the kinetics but alter the ratio of antagonistic to center inputs of scotopic GC responses, and they speed up GCs photopic responses and alter the ratio of GCs' antagonistic to center synaptic inputs and receptive field profiles. (5) Rod bipolar cells shorten response latency of ON GCs and increase the ratio of GCs' antagonistic to center synaptic inputs. (6) Light adaptation speeds up GCs' temporal processing and tunes GC photopic responses to higher frequencies, and the tertiary rod pathway plays a significant role in adaptation-induced TTP changes in some GCs. (7) GC RF center sizes are partially mediated by AIIACs and GC-GC coupling. (8) Connexin36 gap junctions and cone pathways alter synaptic circuits underlying antagonistic surround inputs to GCs in photopic conditions.
Asunto(s)
Luz , Células Fotorreceptoras Retinianas Conos/efectos de la radiación , Células Ganglionares de la Retina/fisiología , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , Vías Visuales/fisiología , Adaptación Ocular , Animales , Visión de Colores/fisiología , Adaptación a la Oscuridad/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Visión Nocturna/fisiología , Tiempo de Reacción , Campos VisualesRESUMEN
Purpose: The purpose of this paper is to describe the extent to which scotopic and photopic measures of visual function predict color fundus photograph (CFP) and fundus autofluorescence (FAF) changes in early and intermediate nonexudative AMD. Methods: Sixty-nine observers were recruited: 56 AMD patients (mean age, 73 ± 12.98 years) and 13 controls (mean age, 67.77 ± 9.72 years). A nonmydriatic retinal camera was used to obtain stereo fundus photographs and FAF images were recorded with a cSLO Heidelberg Spectralis HRA+OCT. Visual acuity (VA) was measured using an Early Treatment of Diabetic Retinopathy Study chart. Contrast sensitivity (CS) was assessed with a Pelli-Robson chart. Dark adaptation (DA) curves were recorded at 3° eccentricity using a PC-based technique. Analysis of these curves yielded five parameters: cone threshold (CT), cone time constant (CC), cone-rod break (α), slope of the second rod component (S2), and rod-rod break (ß). Results: Both cone and rod sensitivity recovery were grossly abnormal in the patients. The rod recovery slope (S2) most accurately predicted the fundus photograph-based grade and the FAF classification (ρ = 0.61 and ρ = 0.60, respectively; both P < 0.0001). CS showed a strong association with FAF (ρ = 0.50, P < 0.0001) and with fundus photograph-based grade (ρ = 0.38, P < 0.002). There was no correlation between VA and either imaging method. Conclusions: Dynamic, rod-based measures most accurately reflect the severity of early AMD. Although less specific to AMD than DA changes, static photopic abnormalities such as CS also correspond with morphologic changes. Assessment of function in early AMD should include dynamic rod- and cone-mediated measurements of sensitivity recovery.
Asunto(s)
Adaptación a la Oscuridad/fisiología , Luz , Degeneración Macular/diagnóstico , Células Fotorreceptoras Retinianas Conos/patología , Células Fotorreceptoras Retinianas Bastones/patología , Agudeza Visual , Anciano , Femenino , Angiografía con Fluoresceína/métodos , Fondo de Ojo , Humanos , Degeneración Macular/fisiopatología , Masculino , Oftalmoscopía , Pronóstico , Recuperación de la Función , Células Fotorreceptoras Retinianas Conos/efectos de la radiación , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , Índice de Severidad de la Enfermedad , Tomografía de Coherencia Óptica/métodosRESUMEN
Purpose: Accumulating evidence suggests that dopamine, the major catecholamine in the vertebrate retina, may modulate cAMP-mediated signaling in photoreceptors to optimize vision in the light/dark cycle. The main putative mechanism of dopamine-induced adaptation changes in photoreceptors is activation of D2-like receptors (D2R), which leads to a decrease of the intracellular cAMP level and reduction of protein kinase A (PKA) activity. However, the mechanisms by which dopamine exerts its regulating effect on the phototransduction cascade remain largely unknown. The aim of the present study was to investigate the effects of dopamine and dopamine receptor agonists on rod photoresponses. Methods: The experiments were performed on solitary rods of the Rana ridibunda frog. Photoreceptor currents were recorded using a suction pipette technique. The effects of dopamine (0.1-50 µM) and selective dopamine receptor agonists-D1R agonist SKF-38393 (0.1-50 µM), D2R agonist quinpirole (2.5-50 µM), and D1-D2 receptor heterodimer agonist SKF-83959 (50 µM)-were examined. Results: We found that, when applied to the rod inner segments (RISs), dopamine and dopamine receptor agonists had no effect on photoresponses. In contrast, the rods responded to dopamine and all agonists applied to their outer segments by decreasing sensitivity to light. At the highest tested concentration (50 µM), the most prominent effect on light sensitivity was induced by D1R agonist SKF-38393, while dopamine, D2R agonist quinpirole, and D1-D2 receptor heterodimer agonist SKF-83959 produced somewhat lower and approximately equal effects. Moreover, SKF-38393 reduced sensitivity at all tested concentrations starting from the smallest one (0.1 µM), whereas dopamine and quinpirole started their action from the higher concentrations of 2.5 µM and 50 µM, respectively. In addition, dopamine, SKF-38393, and quinpirole, on average, did not change the intracellular calcium level as judged from the "exchange current", while SKF-83959 increased it by ~1.3 times. Conclusions: Dopamine induces a decrease in rod sensitivity, mostly by reducing the activation rate of the cascade, and to a much lesser extent, speeding up the turning off of the cascade. The sign of the reaction to all tested drugs, lack of selectivity of dopamine and dopamine receptor agonist action, and analysis of factors that determine sensitivity of photoreceptors suggest that, in rod outer segments (ROSs), dopamine action is mediated by D1-D2 receptor heterodimers but not D1R or D2R alone. This work supports the assumption made earlier by other authors that dopamine exercises its regulatory effect via at least two independent mechanisms, which are cAMP and Ca2+ mediated.
Asunto(s)
Agonistas de Dopamina/farmacología , Dopamina/farmacología , Fototransducción/efectos de los fármacos , Ranidae/fisiología , Receptores de Dopamina D1/agonistas , Células Fotorreceptoras Retinianas Bastones/fisiología , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , 2,3,4,5-Tetrahidro-7,8-dihidroxi-1-fenil-1H-3-benzazepina/farmacología , Animales , Calcio/metabolismo , Cinética , Luz , Fototransducción/efectos de la radiación , Receptores de Dopamina D1/metabolismo , Segmento Externo de la Célula en Bastón/efectos de los fármacos , Segmento Externo de la Célula en Bastón/metabolismo , Segmento Externo de la Célula en Bastón/efectos de la radiación , Factores de TiempoRESUMEN
Purpose: To study rod- and cone-driven adaptation dynamics separately, we used the silent substitution technique to selectively stimulate rods or cones in the Opn1lwLIAIS (LIAIS) mouse, in which the native M-cone pigment is replaced by a human L-cone pigment (L*). Methods: ERG recordings were performed on anesthetized LIAIS mice. ERG stimuli were sinusoidally modulated. After 10 minutes of adaptation to 0.4 candela per square meter (cd/m2) ERGs were measured, followed by 11-minute adaptation to 8.8 cd/m2 background and recordings directly after the luminance increase and every second minute. Finally, during adaptation to 0.4 cd/m2 for 32 minutes, ERG responses were recorded directly after the change in background and every second minute. This protocol was repeated with rod-isolating stimuli (8 Hz; 75% rod contrast), L*-cone-isolating stimuli (12 Hz; 55% cone contrast) and white light (8 Hz and 12 Hz; 100% Michelson contrast). Results: At 8.8 cd/m2, responses directly displayed photopic response properties without further changes in either cone or white light responses. Rod-driven responses were very small. After the return to 0.4 cd/m2, both rod-driven and white light responses increased over a time course of about 30 minutes. Cone-driven responses were very small. Response phases changed directly after a change in background without further alterations. Conclusions: Rod- and cone-driven signal pathways display strongly different adaptation characteristics: adaptation of cone-driven responses to photopic conditions is very fast, whereas rod-driven responses change with a time course up to 30 minutes during scotopic conditions. Luminance responses are cone-driven at 8.8 cd/m2 and rod-driven at 0.4 cd/m2.
Asunto(s)
Adaptación Ocular/fisiología , Células Fotorreceptoras Retinianas Conos , Células Fotorreceptoras Retinianas Bastones , Animales , Electrorretinografía , Ratones , Ratones Endogámicos C57BL , Estimulación Luminosa/métodos , Células Fotorreceptoras Retinianas Conos/fisiología , Células Fotorreceptoras Retinianas Conos/efectos de la radiación , Células Fotorreceptoras Retinianas Bastones/fisiología , Células Fotorreceptoras Retinianas Bastones/efectos de la radiaciónRESUMEN
We previously reported autophagic structures in rod photoreceptors expressing a misfolding RHO (rhodopsin) mutant (RHOP23H), suggesting that autophagy may play a role in degrading the mutant RHO and/or be involved in photoreceptor cell death. To further examine autophagy in normal and diseased rods, we generated transgenic Xenopus laevis tadpoles expressing the dually fluorescent autophagy marker mRFP-eGFP-LC3 in rods, which changes from green to yellow and finally red as autophagic structures develop and mature. Using transgenic lines with constitutive and inducible expression, we determined the time-course of autophagy in rod photoreceptors: autophagosomes last for 6 to 8 hours before fusing with lysosomes, and acidified autolysosomes last for about 28 hours before being degraded. Autophagy was diurnally regulated in normal rods, with more autophagic structures generated during periods of light, and this regulation was non-circadian. We also found that more autophagosomes were produced in rods expressing the misfolding RHOP23H mutant. The RHO chromophore absorbs photons to initiate phototransduction, and is consumed in this process; it also promotes RHO folding. To determine whether increased autophagy in light-exposed normal rods is caused by increased RHO misfolding or phototransduction, we used CRISPR/Cas9 to knock out the RPE65 and GNAT1 genes, which are essential for chromophore biosynthesis and phototransduction respectively. Both knockouts suppressed light-induced autophagy, indicating that although light and misfolded rhodopsin can both induce autophagy in rods, light-induced autophagy is not due to misfolding of RHO, but rather due to phototransduction. Abbreviations: CYCS: cytochrome c; bRHOP23H: bovine RHOP23H; Cas9: CRISPR associated protein 9; dpf: days post-fertilization; eGFP: enhanced green fluorescent protein; GNAT1: guanine nucleotide-binding protein G(t) subunit alpha-1 aka rod alpha-transducin; HSPA1A/hsp70: heat shock protein of 70 kilodaltons; LAMP1: lysosomal-associated membrane protein 1; LC3: microtubule-associated protein 1A/1B light chain 3; mRFP: monomeric red fluorescent protein; RHO: rhodopsin; RP: retinitis pigmentosa; RPE65: retinal pigment epithelium-specific 65 kDa protein: sfGFP: superfolding GFP; sgRNA: single guide RNA; WGA: wheat germ agglutinin; RHOp: the Xenopus laevis RHO.2.L promoter.
Asunto(s)
Autofagia/genética , Autofagia/efectos de la radiación , Fototransducción/genética , Células Fotorreceptoras Retinianas Bastones/metabolismo , Retinitis Pigmentosa/metabolismo , Rodopsina/metabolismo , Animales , Animales Modificados Genéticamente , Autofagosomas/metabolismo , Autofagosomas/efectos de la radiación , Ritmo Circadiano/genética , Ritmo Circadiano/efectos de la radiación , Fluorescencia , Proteínas Fluorescentes Verdes/química , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Proteínas de Unión al GTP Heterotriméricas/genética , Proteínas de Unión al GTP Heterotriméricas/metabolismo , Larva/genética , Larva/metabolismo , Larva/ultraestructura , Fototransducción/efectos de la radiación , Mutación , Células Fotorreceptoras Retinianas Bastones/citología , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , Células Fotorreceptoras Retinianas Bastones/ultraestructura , Retinitis Pigmentosa/genética , Rodopsina/química , Rodopsina/genética , Rodopsina/efectos de la radiación , Factores de Tiempo , Xenopus laevis , cis-trans-Isomerasas/genética , cis-trans-Isomerasas/metabolismoRESUMEN
OBJECTIVE: To review clinical and pre-clinical evidence supporting the role of visual pathways, from the eye to the cortex, in the development of photophobia in headache disorders. BACKGROUND: Photophobia is a poorly understood light-induced phenomenon that emerges in a variety of neurological and ophthalmological conditions. Over the years, multiple mechanisms have been proposed to explain its causes; however, scarce research and lack of systematic assessment of photophobia in patients has made the search for answers quite challenging. In the field of headaches, significant progress has been made recently on how specific visual networks contribute to photophobia features such as light-induced intensification of headache, increased perception of brightness and visual discomfort, which are frequently experienced by migraineurs. Such progress improved our understanding of the phenomenon and points to abnormal processing of light by both cone/rod-mediated image-forming and melanopsin-mediated non-image-forming visual pathways, and the consequential transfer of photic signals to multiple brain regions involved in sensory, autonomic and emotional regulation. CONCLUSION: Photophobia phenotype is diverse, and the relative contribution of visual, trigeminal and autonomic systems may depend on the disease it emerges from. In migraine, photophobia could result from photic activation of retina-driven pathways involved in the regulation of homeostasis, making its association with headache more complex than previously thought.
Asunto(s)
Cefalea/fisiopatología , Fotofobia/fisiopatología , Vías Visuales/fisiopatología , Animales , Ceguera/fisiopatología , Tronco Encefálico/fisiopatología , Color , Cefalea/complicaciones , Humanos , Luz/efectos adversos , Mesencéfalo/fisiopatología , Ratones , Trastornos Migrañosos/complicaciones , Trastornos Migrañosos/fisiopatología , Estimulación Luminosa/efectos adversos , Fotofobia/etiología , Células Ganglionares de la Retina/fisiología , Células Fotorreceptoras Retinianas Bastones/fisiología , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , Opsinas de Bastones/fisiología , Corteza Somatosensorial/fisiopatología , Tálamo/fisiopatologíaRESUMEN
Retinal dopamine is a critical modulator of high acuity, light-adapted vision and photoreceptor coupling in the retina. Dopaminergic amacrine cells (DACs) serve as the sole source of retinal dopamine, and dopamine release in the retina follows a circadian rhythm and is modulated by light exposure. However, the retinal circuits through which light influences the development and function of DACs are still unknown. Intrinsically photosensitive retinal ganglion cells (ipRGCs) have emerged as a prime target for influencing retinal dopamine levels because they costratify with DACs in the inner plexiform layer and signal to them in a retrograde manner. Surprisingly, using genetic mouse models lacking specific phototransduction pathways, we find that while light influences the total number of DACs and retinal dopamine levels, this effect does not require ipRGCs. Instead, we find that the rod pathway is a critical modulator of both DAC number and retinal dopamine levels.
Asunto(s)
Células Amacrinas/citología , Células Amacrinas/efectos de la radiación , Dopamina/metabolismo , Luz , Células Amacrinas/metabolismo , Animales , Recuento de Células , Femenino , Fototransducción , Masculino , Ratones , Modelos Biológicos , Células Fotorreceptoras Retinianas Conos/metabolismo , Células Fotorreceptoras Retinianas Conos/efectos de la radiación , Células Ganglionares de la Retina/metabolismo , Células Ganglionares de la Retina/efectos de la radiación , Células Fotorreceptoras Retinianas Bastones/metabolismo , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , Opsinas de Bastones/metabolismo , Tirosina 3-Monooxigenasa/metabolismoRESUMEN
Vision is mediated by two types of photoreceptors: rods, enabling vision in dim light; and cones, which function in bright light. Despite many similarities in the components of their respective phototransduction cascades, rods and cones have distinct sensitivity, response kinetics, and adaptation capacity. Cones are less sensitive and have faster responses than rods. In addition, cones can function over a wide range of light conditions whereas rods saturate in moderately bright light. Calcium plays an important role in regulating phototransduction and light adaptation of rods and cones. Notably, the two dominant Ca2+-feedbacks in rods and cones are driven by the identical calcium-binding proteins: guanylyl cyclase activating proteins 1 and 2 (GCAPs), which upregulate the production of cGMP; and recoverin, which regulates the inactivation of visual pigment. Thus, the mechanisms producing the difference in adaptation capacity between rods and cones have remained poorly understood. Using GCAPs/recoverin-deficient mice, we show that mammalian cones possess another Ca2+-dependent mechanism promoting light adaptation. Surprisingly, we also find that, unlike in mouse rods, a unique Ca2+-independent mechanism contributes to cone light adaptation. Our findings point to two novel adaptation mechanisms in mouse cones that likely contribute to the great adaptation capacity of cones over rods.
Asunto(s)
Adaptación Ocular/fisiología , Calcio/metabolismo , Luz , Células Fotorreceptoras Retinianas Conos/efectos de la radiación , Animales , GMP Cíclico/metabolismo , Proteínas Activadoras de la Guanilato-Ciclasa/deficiencia , Proteínas Activadoras de la Guanilato-Ciclasa/genética , Proteínas Activadoras de la Guanilato-Ciclasa/metabolismo , Iones/química , Cinética , Ratones , Ratones Noqueados , Recoverina/deficiencia , Recoverina/genética , Recoverina/metabolismo , Células Fotorreceptoras Retinianas Conos/metabolismo , Células Fotorreceptoras Retinianas Bastones/metabolismo , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , Regulación hacia Arriba/efectos de la radiaciónRESUMEN
Stimulus- or context-dependent routing of neural signals through parallel pathways can permit flexible processing of diverse inputs. For example, work in mouse shows that rod photoreceptor signals are routed through several retinal pathways, each specialized for different light levels. This light-level-dependent routing of rod signals has been invoked to explain several human perceptual results, but it has not been tested in primate retina. Here, we show, surprisingly, that rod signals traverse the primate retina almost exclusively through a single pathway - the dedicated rod bipolar pathway. Identical experiments in mouse and primate reveal substantial differences in how rod signals traverse the retina. These results require reevaluating human perceptual results in terms of flexible computation within this single pathway. This includes a prominent speeding of rod signals with light level - which we show is inherited directly from the rod photoreceptors themselves rather than from different pathways with distinct kinetics.
Asunto(s)
Células Fotorreceptoras Retinianas Bastones/metabolismo , Transducción de Señal , Células Amacrinas/metabolismo , Células Amacrinas/efectos de la radiación , Animales , Cinética , Luz , Fototransducción/efectos de la radiación , Macaca , Ratones Endogámicos C57BL , Estimulación Luminosa , Células Bipolares de la Retina/metabolismo , Células Bipolares de la Retina/efectos de la radiación , Células Fotorreceptoras Retinianas Conos/metabolismo , Células Fotorreceptoras Retinianas Conos/efectos de la radiación , Células Ganglionares de la Retina/metabolismo , Células Ganglionares de la Retina/efectos de la radiación , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , Relación Señal-RuidoRESUMEN
Rhodopsin homeostasis is tightly coupled to rod photoreceptor cell survival and vision. Mutations resulting in the misfolding of rhodopsin can lead to autosomal dominant retinitis pigmentosa (adRP), a progressive retinal degeneration that currently is untreatable. Using a cell-based high-throughput screen (HTS) to identify small molecules that can stabilize the P23H-opsin mutant, which causes most cases of adRP, we identified a novel pharmacological chaperone of rod photoreceptor opsin, YC-001. As a non-retinoid molecule, YC-001 demonstrates micromolar potency and efficacy greater than 9-cis-retinal with lower cytotoxicity. YC-001 binds to bovine rod opsin with an EC50 similar to 9-cis-retinal. The chaperone activity of YC-001 is evidenced by its ability to rescue the transport of multiple rod opsin mutants in mammalian cells. YC-001 is also an inverse agonist that non-competitively antagonizes rod opsin signaling. Significantly, a single dose of YC-001 protects Abca4 -/- Rdh8 -/- mice from bright light-induced retinal degeneration, suggesting its broad therapeutic potential.
Asunto(s)
Fármacos Neuroprotectores/farmacología , Pliegue de Proteína/efectos de los fármacos , Degeneración Retiniana/tratamiento farmacológico , Células Fotorreceptoras Retinianas Bastones/efectos de los fármacos , Rodopsina/metabolismo , Tiofenos/farmacología , Transportadoras de Casetes de Unión a ATP/genética , Oxidorreductasas de Alcohol/genética , Animales , Línea Celular Tumoral , Modelos Animales de Enfermedad , Diterpenos , Femenino , Células HEK293 , Ensayos Analíticos de Alto Rendimiento , Humanos , Luz/efectos adversos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Mutación , Células 3T3 NIH , Fármacos Neuroprotectores/uso terapéutico , Transporte de Proteínas/efectos de los fármacos , Transporte de Proteínas/genética , Degeneración Retiniana/etiología , Degeneración Retiniana/patología , Células Fotorreceptoras Retinianas Bastones/metabolismo , Células Fotorreceptoras Retinianas Bastones/patología , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , Retinaldehído/farmacología , Retinaldehído/uso terapéutico , Rodopsina/agonistas , Rodopsina/antagonistas & inhibidores , Rodopsina/genética , Tiofenos/uso terapéutico , Resultado del TratamientoRESUMEN
Vision in dim light depends on synapses between rods and rod bipolar cells (RBCs). Here, we find that these synapses exist in multiple configurations, in which single release sites of rods are apposed by one to three postsynaptic densities (PSDs). Single RBCs often form multiple PSDs with one rod; and neighboring RBCs share ~13% of their inputs. Rod-RBC synapses develop while ~7% of RBCs undergo programmed cell death (PCD). Although PCD is common throughout the nervous system, its influences on circuit development and function are not well understood. We generate mice in which ~53 and ~93% of RBCs, respectively, are removed during development. In these mice, dendrites of the remaining RBCs expand in graded fashion independent of light-evoked input. As RBC dendrites expand, they form fewer multi-PSD contacts with rods. Electrophysiological recordings indicate that this homeostatic co-regulation of neurite and synapse development preserves retinal function in dim light.
Asunto(s)
Homeostasis , Plasticidad Neuronal/fisiología , Sinapsis/fisiología , Vías Visuales/fisiología , Animales , Axones/metabolismo , Recuento de Células , Dendritas/fisiología , Dendritas/efectos de la radiación , Homeostasis/efectos de la radiación , Luz , Ratones Transgénicos , Plasticidad Neuronal/efectos de la radiación , Células Bipolares de la Retina/citología , Células Bipolares de la Retina/efectos de la radiación , Células Fotorreceptoras Retinianas Bastones/citología , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , Sinapsis/efectos de la radiación , Vías Visuales/efectos de la radiaciónRESUMEN
The conventional view was that cones are responsible for pupil constriction in photopic lighting conditions. With the discovery of intrinsically photosensitive retinal ganglion cells (ipRGC), it was found that signals from ipRGCs along with cones mediated the pupil light reflex in photopic lighting conditions. Although both signals contributed, it was unclear how these signals were summed. In the work reported here, steady-state pupil size was measured with an infrared camera under LED lighting conditions with different color temperatures and luminance. A formula was then derived for pupil size according to the linear summation of cirtopic and photopic luminance. This formula allowed direct calculations to predict pupil size well when LED photopic luminance ranged from about 50cd/m2 to 300cd/m2, which is the general luminance level range for computer and smartphone screens.
Asunto(s)
Visión de Colores/fisiología , Estimulación Luminosa , Pupila/fisiología , Reflejo Pupilar/fisiología , Células Fotorreceptoras Retinianas Conos/efectos de la radiación , Células Ganglionares de la Retina/efectos de la radiación , Células Fotorreceptoras Retinianas Bastones/efectos de la radiación , Adolescente , Femenino , Humanos , Masculino , Adulto JovenRESUMEN
Hallmarks of Retinitis Pigmentosa (RP), a family of genetic diseases, are a typical rod-cone-degeneration with initial night blindness and loss of peripheral vision, followed by decreased daylight sight and progressive visual acuity loss up to legal blindness. Great heterogeneity in nature and function of mutated genes, variety of mutations for each of them, variability in phenotypic appearance and transmission modality contribute to make RP a still incurable disease. Translational research relies on appropriate animal models mimicking the genetic and phenotypic diversity of the human pathology. Here, we provide a systematic, morphological and functional analysis of RhoTvrm4/Rho+ rhodopsin mutant mice, originally described in 2010 and portraying several features of common forms of autosomal dominant RP caused by gain-of-function mutations. These mice undergo photoreceptor degeneration only when exposed briefly to strong, white light and allow controlled timing of induction of rod and cone death, which therefore can be elicited in adult animals, as observed in human RP. The option to control severity and retinal extent of the phenotype by regulating intensity and duration of the inducing light opens possibilities to exploit this model for multiple experimental purposes. Altogether, the unique features of this mutant make it an excellent resource for retinal degeneration research.
Asunto(s)
Modelos Animales de Enfermedad , Retina/patología , Retina/fisiología , Retinitis Pigmentosa/patología , Rodopsina/genética , Rodopsina/metabolismo , Animales , Supervivencia Celular/efectos de la radiación , Luz , Ratones , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Células Fotorreceptoras Retinianas Conos/fisiología , Células Fotorreceptoras Retinianas Conos/efectos de la radiación , Células Fotorreceptoras Retinianas Bastones/fisiología , Células Fotorreceptoras Retinianas Bastones/efectos de la radiaciónRESUMEN
The light absorbing chromophore in opsin visual pigments is the protonated Schiff base of 11-cis-retinaldehyde (11cRAL). Absorption of a photon isomerizes 11cRAL to all-trans-retinaldehyde (atRAL), briefly activating the pigment before it dissociates. Light sensitivity is restored when apo-opsin combines with another 11cRAL to form a new visual pigment. Conversion of atRAL to 11cRAL is carried out by enzyme pathways in neighboring cells. Here we show that blue (450-nm) light converts atRAL specifically to 11cRAL through a retinyl-phospholipid intermediate in photoreceptor membranes. The quantum efficiency of this photoconversion is similar to rhodopsin. Photoreceptor membranes synthesize 11cRAL chromophore faster under blue light than in darkness. Live mice regenerate rhodopsin more rapidly in blue light. Finally, whole retinas and isolated cone cells show increased photosensitivity following exposure to blue light. These results indicate that light contributes to visual-pigment renewal in mammalian rods and cones through a non-enzymatic process involving retinyl-phospholipids.It is currently thought that visual pigments in vertebrate photoreceptors are regenerated exclusively through enzymatic cycles. Here the authors show that mammalian photoreceptors also regenerate opsin pigments in light through photoisomerization of N-ret-PE (N-retinylidene-phosphatidylethanolamine.