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This study investigates deep-learning (DL) sequence modeling techniques to reliably fit dark adaptation (DA) curves and estimate their key parameters in patients with age-related macular degeneration (AMD) to improve robustness and curve predictions.A long-short-term memory autoencoder was used as the DL method to model the DA curve. The performance was compared against the classical nonlinear regression method using goodness-of-fit and repeatability metrics. Experiments were performed to predict the latter portion of the curve using data from early measurements. The prediction accuracy was quantified as the rod intercept time (RIT) prediction error between predicted and actual curves.The two models had comparable goodness-of-fit measures, with root mean squared error (RMSE; SD) = 0.11 (0.04) log-units (LU) for the classical model and RMSE = 0.13 (0.06) LU for the DL model. Repeatability of the curve fits evaluated after introduction of random perturbations, and after performing repeated testing, demonstrated superiority of the DL method, especially among parameters related to cone decay. The DL method exhibited superior ability to predict the curve and RIT using points prior to -2 LU, with 3.1 ± 3.1 minutes RIT prediction error, compared to 19.1 ± 18.6 minutes RIT error for the classical method.The parameters obtained from the DL method demonstrated superior robustness as well as predictability of the curve. These could provide important advances in using multiple DA curve parameters to characterize AMD severity.Dark adaptation is an important functional measure in studies of AMD and curve modeling using DL methods can lead to improved clinical trial end points.

Purpose To determine whether dilation status has a clinically meaningful effect on sensitivity in normal subjects undergoing two-color dark-adapted perimetry, which can be useful to assess rod function. Methods A perimeter measured naturally and pharmacologically dilated scotopic sensitivities using a test grid consisting of 16 points across the horizontal meridian ranging from 60° temporal to 45° nasal using cyan (500 nm wavelength) or red (650 nm wavelength) stimuli. The primary outcome was average overall sensitivity based on dilation status, which was compared using a linear mixed effect model for each color stimuli. A difference of 2 dB or more was considered clinically significant. Results Twenty-nine eyes from 15 subjects (nine female) ages 23 to 63 with no known retinal pathology were included. Pharmacologically dilated eyes were 0.54 dB (95% confidence interval [CI], 0.05 dB to 1.03 dB; P = 0.032) more sensitive to a red stimulus than naturally dilated eyes, but this was not statistically significant after correction for multiple comparisons. Pharmacologically dilated eyes were 0.03 dB (95% CI, −0.20 dB to 0.14 dB; P = 0.734) less sensitive to a cyan stimulus compared to naturally dilated eyes. Conclusions These findings show no clinically significant differences in sensitivity of scotopic perimetry in eyes without retinal pathology based on dilation status for both cyan and red stimuli. Translational Relevance In this study, pharmacological dilation did not have a clinically meaningful effect on sensitivity, suggesting that this is not necessary when using two-color dark-adapted perimetry to assess for rod function.

Purpose The purpose of this study was to characterize changes in the full-field flash electroretinogram (ERG) in association with psychophysical dark-adapted visual thresholds in patients with genetically characterized Duchenne muscular dystrophy (DMD) either lacking Dp427 (Up 30) or at least Dp260 in addition to Dp427 (Down 30). Methods Twenty-one patients with DMD and 27 age-similar controls participated in this study. Dark-adapted (0.01, 3.0, and 10 cd.s/m² flashes) and light-adapted (3.0 cd.s/m² flash) ERGs were recorded following International Society for Clinical Electrophysiology of Vision (ISCEV) standard protocols. Visual detection thresholds to 625-nm (cone function) and 527-nm (rod function) light-emitting diode (LED) flashes (2 degree diameter) were measured during a dark adaptation period after a 1-minute exposure to a bleaching light (3000 cd/m²). Initially, 8 minutes of interleaved 625-nm and 527-nm thresholds were measured. After an additional 5 minutes of dark-adaptation, a second set of threshold measurements to 527-nm stimuli was performed during the subsequent 6 minutes. Results Dark-adapted b-wave amplitude was significantly reduced to all strengths of flash and a-wave in response to the strong flash stimulus was delayed (15.6 vs. 14.7 ms, P < 0.05) in patients with Down 30 compared with controls. Dark-adapted cone thresholds did not differ among the groups (−2.0, −1.8, and −1.7 log cd/m² for Down 30, Up 30, and controls, respectively, P = 0.21). In contrast, dark-adapted rod thresholds were elevated (F(2,36) = 8.537, P = 0.001) in patients with Down 30 (mean = −3.2 ± 1.1 log cd/m²) relative to controls (mean = −4.2 ± 0.3 log cd/m²). Dark-adapted b-wave amplitudes were correlated with dark-adapted rod sensitivity in patients with DMD (Spearman Rho = 0.943, P = 0.005). The changes were much smaller or absent in patients with intact Dp260. Conclusions Dp260 is particularly required for normal rod-system function in dark adaptation.

When scanning across a scene, luminance can vary by up to 100,000-to-1 (high dynamic range, HDR), requiring multiple normalizing mechanisms spanning from the retina to the cortex to support visual acuity and recognition. Vision models based on standard dynamic range (SDR) luminance contrast ratios below 100-to-1 have limited ability to generalize to real-world scenes with HDR luminance. To characterize how orientation and luminance are linked in brain mechanisms for luminance normalization, we measured orientation discrimination of Gabor targets under HDR luminance dynamics. We report a novel phenomenon, that abrupt 10- to 100-fold darkening engages contextual facilitation, distorting the apparent orientation of a high-contrast central target. Surprisingly, facilitation was influenced by grouping by luminance similarity, as well as by the degree of luminance variability in the surround. These results challenge vision models based solely on activity normalization and raise new questions that will lead to models that perform better in real-world scenes.

Purpose To evaluate the nature and extent of functional abnormality in X-linked retinoschisis (XLRS) by comparing three dark-adapted, full-field measures: the electroretinogram (ERG), pupillary light reflex (PLR), and luminance threshold. Methods ERGs, PLRs (pupil constriction due to light stimulation), and luminance thresholds were measured from seven XLRS subjects and from 10 normally sighted, age-similar controls. ERGs and PLRs were obtained for a range of flash strengths, and these data were fit with Naka–Rushton functions to derive the maximum saturated b-wave (Vmax) and PLR (Pmax) amplitudes. Additionally, semi-saturation constants were obtained for the b-wave (σ) and PLR (s). Values of 1/σ and 1/s provide sensitivity measures. Full-field, dark-adapted luminance thresholds were measured using 465-nm and 642-nm flash stimuli. Results V max and 1/σ were significantly reduced in XLRS compared to the controls (both t ≥ 5.33, P < 0.001). In comparison, Pmax was normal in the XLRS subjects (t = 1.39, P = 0.19), but 1/s was reduced (t = 7.84, P < 0.001). Luminance thresholds for the control and XLRS groups did not differ significantly (F = 3.57, P = 0.08). Comparisons among measures indicated that pupil sensitivity was correlated with luminance threshold for the long- and short-wavelength stimuli (both, r ≥ 0.77, P ≤ 0.04). Correlations among all other measures were not statistically significant. Conclusions The results indicate that the presumed bipolar cell dysfunction in XLRS, indicated by b-wave abnormalities, has complex downstream effects: Dark-adapted luminance threshold and maximum pupil responses are not significantly affected, but pupil sensitivity is reduced.

Purpose The purpose of this study was to investigate the acute effects of caffeine on retinal hemodynamics during dark to light adaptation using optical coherence tomography angiography (OCTA). Methods Thirteen healthy individuals (13 eyes) underwent OCTA imaging after dark adaptation and at repeated intervals during the transition to ambient light in two imaging sessions: control and after ingesting 200 mg of caffeine. We analyzed the parafoveal vessel density (VD) and adjusted flow index (AFI) of the superficial capillary plexus (SCP), middle capillary plexus (MCP), and deep capillary plexus (DCP), as well as the vessel length density (VLD) of the SCP. After adjusting for age, refractive error, and scan quality, we compared parameters between control and caffeine conditions. Results In the dark, MCP VD decreased significantly after caffeine (-2.63 ± 1.28%). During the transition to light, initially, DCP VD increased (12.55 ± 2.52%), whereas SCP VD decreased (-2.09 ± 0.91%) significantly with caffeine compared to control. By 15 minutes in light, DCP VD reversed and was significantly decreased (-5.45 ± 2.62%), whereas MCP VD increased (4.65 ± 1.74%). There were no differences in AFI or VLD. Conclusions We show that, overall, caffeine causes a trend of delayed vascular response in all three macular capillary plexuses in response to ambient light. Whereas the MCP is constricted in the dark, during the transition from dark to light, there is initially delay followed by prolonged constriction of the DCP and constriction followed by slow dilation of the SCP. We posit that these delayed vascular responses may present potential risk of capillary ischemia.

Purpose To assess which visual function measures are most strongly associated with overall retinal drusen volume in age-related macular degeneration (AMD). Methods A total of 100 eyes (16 eyes with early AMD, 62 eyes with intermediate AMD, and 22 eyes from healthy controls) were recruited in this cross-sectional study. All subjects underwent several functional assessments: best-corrected visual acuity (BCVA), low-luminance visual acuity (LLVA), visual acuity (VA) measured with the Moorfields Acuity Chart (MAC-VA), contrast sensitivity with the Pelli–Robson test, reading speed using the International Reading Speed texts, and mesopic and dark-adapted microperimetry. Drusen volume was automatically determined based on optical coherence tomography using an approach based on convolutional neural networks. The relationship between drusen volume and visual function was assessed with linear regressions controlling for confounders. Results Mean drusen volume and MAC-VA differed significantly among all AMD stages and controls (P < 0.001). In univariate linear regression, LLVA, MAC-VA, contrast sensitivity, and mesopic and dark-adapted microperimetry were significantly negatively associated with the overall drusen volume (all P < 0.006). After controlling for AMD stage, age, and the presence of subretinal drusenoid deposits, MAC-VA and mesopic and dark-adapted microperimetry were still significantly associated with drusen volume (P = 0.008, P = 0.023, and P = 0.022, respectively). Conclusions Our results suggest that MAC-VA, as well as mesopic and dark-adapted microperimetry, might indicate structural changes related to drusen volume in early stages of AMD.

Administration of voriconazole, an antifungal triazole, causes transient visual disturbances in patients and attenuates the b-wave of the ERG. We sought to identify the retinal target of voriconazole underlying the effect on the ERG b-wave.Electroretinograms were recorded from mice before and after intraperitoneal injection of voriconazole. The effect of voriconazole on ON-bipolar cells was tested by patch-clamp recordings of ON-bipolar cells in mouse retinal slices. Effects of voriconazole on mGluR6 and TRPM3 were assessed by patch-clamp recordings of Chinese hamster ovary (CHO) and HEK293 cells transfected with either TRPM3 or mGluR6 plus Kir3.1/Kir3.4.Voriconazole attenuated the ERG b-wave in mice, and inhibited ON-bipolar cell responses evoked by application of CPPG, an mGluR6 antagonist, onto the ON-bipolar cell dendrites, indicating that voriconazole blocks a step in the mGluR6-TRPM1 signal transduction pathway. Voriconazole almost completely blocked capsaicin-activated currents in ON-bipolar cells, which have been attributed to direct activation of the TRPM1 cation channel. Furthermore, application of voriconazole to CHO cells expressing TRPM3, a closely related channel to TRPM1, showed that voriconazole reversibly blocked pregnenolone sulfate-stimulated TRPM3 currents in transfected cells. In contrast, voriconazole only slightly inhibited mGluR6-mediated activation of G-protein activated inward rectifier potassium (GIRK) currents in cotransfected cells, suggesting that mGluR6 is not the primary target of voriconazole in ON-bipolar cells.The visual disturbances associated with voriconazole are likely due to block of TRPM1 channels in retinal ON-bipolar cells. Other neurological effects of voriconazole may be due to block of TRPM3 channels expressed in the brain.

PURPOSE We evaluated the test-retest reliability of current methods of inducing the melanopsin-driven postillumination pupil response (PIPR) under hemifield, central-field, and full-field stimulation conditions. METHODS Pupil response was recorded with an eye tracker in 10 visually normal participants. Light stimuli were presented using a Ganzfeld screen with a custom-built device that allows specific regions of the retina to be stimulated. Blue light stimulation at 400 cd/m(2) intensity was presented for 400 ms to the lower and upper halves of the central 30° fields (hemifields), central 30° field (central-field), and full-field to induce PIPR. Red light full-field stimulation also was presented with the same intensity and duration as a control condition. Test-retest reliability of the PIPR measures was assessed by calculating the intraclass correlation coefficient (ICC) of six repetitions for lower and upper hemifield stimulation, and three repetitions for central-field and full-field stimulation. RESULTS Hemifield, central-field, and full-field blue light stimulation induced increasingly greater PIPR in ascending order, while full-field red light stimulation induced no PIPR. Mean lower and upper hemifield PIPR was highly symmetric. Mean ICC of blue light PIPR was 0.87 for lower hemifield, 0.88 for upper hemifield, 0.95 for central-field, and 0.94 for full-field stimulation. CONCLUSIONS We validated a new and repeatable method to measure PIPR induced by hemifield, central-field, and full-field light stimulation. Good PIPR measurement reliability was obtained under all conditions. This practical and reliable protocol will facilitate the clinical application of PIPR testing in different disease populations.

PURPOSE To assess temporal summation in children with a history of retinopathy of prematurity (ROP) by determining the critical duration (tCRIT) for complete temporal summation under rod-mediated conditions. From prior ERG studies, it is known that the kinetics of activation of phototransduction are prolonged in the ROP rod photoreceptor. METHODS Dark-adapted thresholds for detecting 10° diameter stimuli with durations from 10 to 640 ms were measured. A two-alternative, spatial, forced-choice psychophysical procedure was used. The tCRIT for complete summation was estimated in former preterm subjects with a history of severe ROP (n = 7), mild ROP (n = 23), and no ROP (n = 15). The subjects ranged in age from 10.4 to 17.6 (median 15.6) years. Age-similar term-born control subjects (n = 5) were also tested. RESULTS Critical duration was significantly longer in subjects with a history of ROP than in subjects who never had ROP or who were born at term. Mean tCRIT in the mild ROP group [127.5 (SD = 19.9) ms] and severe group [147.6 (SD = 18.9) ms] did not differ significantly, but both were significantly longer than in former preterms who never had ROP [101.1 (SD = 16.5) ms] and in term-born controls [101.0 (SD = 19.5) ms]. CONCLUSIONS In ROP subjects, tCRIT is significantly prolonged. This is likely due to abnormal kinetics in the rod outer segment.

PURPOSE: We estimate and compare change in iris cross-sectional area (IA) after physiologic and pharmacologic mydriasis in subjects with different dominant mechanisms for primary angle closure. METHODS: Anterior segment optical coherence tomography (AS-OCT) measurements in light, dark, and following pharmacologic dilation were obtained on primary angle closure suspects (PACS), primary angle closure (PAC), and primary angle closure glaucoma (PACG) examined during the 5-year follow-up of the Handan Eye Study. Subjects were categorized into three subgroups according to their dominant angle closure (AC) mechanisms as determined by AS-OCT: pupillary block (PB), plateau iris configuration (PIC), and thick peripheral iris roll (TPIR). The IA and other biometric parameters measured using the Zhongshan Angle Assessment Program in the right eyes of all subjects were analyzed. RESULTS: A total of 364 right eyes of subjects with PACS (333), and PAC/PACG (31) was included in the analysis. Significant differences in the change of IAs (P = 0.030), IA loss per mm pupil diameter (PD) increase (P = 0.001) in light versus pharmacologic dilation, and IA loss per mm PD increase (P = 0.011) from dark versus pharmacologic dilation were observed among the three groups. The smallest decrease occurred in the PB group. CONCLUSIONS: There are significant differences in IA and IA loss per mm of pupil change following physiologic or pharmacologic mydriasis in Chinese subjects with dissimilar dominant mechanisms for AC. Dynamic iris change may have a more important role in angle closure where PB is the dominant mechanism.

The staircase method has been widely used in measuring perceptual learning. Recently, Zhao, Lesmes, and Lu (2017, 2019) developed the quick Change Detection (qCD) method and applied it to measure the trial-by-trial time course of dark adaptation. In the current study, we conducted two simulations to evaluate the performance of the 3-down/1-up staircase and qCD methods in measuring perceptual learning in a two-alternative forced-choice task. In Study 1, three observers with different time constants (40, 80, and 160 trials) of an exponential learning curve were simulated. Each simulated observer completed staircases with six step sizes (1%, 5%, 10%, 20%, 30%, and 60%) and a qCD procedure, each starting at five levels (+50%, +25%, 0, -25%, and -50% different from the true threshold in the first trial). We found the following results: Staircases with 1% and 5% step sizes failed to generate more than five reversals half of the time; and the bias and standard deviations of thresholds estimated from the post hoc segment-by-segment qCD analysis were much smaller than those from the staircase method with the other four step sizes. In Study 2, we simulated thresholds in the transfer phases with the same time constants and 50% transfer for each observer in Study 1. We found that the estimated transfer indexes from qCD showed smaller biases and standard deviations than those from the staircase method. In addition, rescoring the simulated data from the staircase method using the Bayesian estimation component of the qCD method resulted in much-improved estimates. We conclude that the qCD method characterizes the time course of perceptual learning and transfer more accurately, precisely, and efficiently than the staircase method, even with the optimal 10% step size.

Exposure of kittens to complete darkness for 10 days has been shown (Duffy & Mitchell, 2013) to reverse the loss of visual acuity that follows a prior period of monocular deprivation (MD). In that study, recovery of acuity in the previously deprived eye was fast despite the fact that darkness was imposed 2 months after the period of MD when kittens were 3 months old. In a later study (Holman, Duffy, & Mitchell, 2018), it was demonstrated that the same period of darkness was ineffective when it was imposed on cats about 1 year old, suggesting that dark exposure may only promote recovery when applied within an early critical period. To determine the profile of this critical period, the identical period of darkness (10 days) was imposed on kittens at various ages that had all received the same 7-day period of MD from postnatal day 30 (P30). Recovery of the acuity of the deprived eye as measured by use of a jumping stand was complete when darkness was imposed prior to P186 days, but thereafter, darkness induced progressively smaller acuity improvements and was ineffective in kittens when it began at or beyond P191 days of age. These data indicate a critical period for darkness-induced recovery with an abrupt end over a 5-day period.

Purpose To test the hypothesis that oxidative stress in the outer retina (OR = distance from external limiting membrane to the retinal pigment epithelium–choroid boundary) can be detected by using antioxidants (AOs) to correct an impaired light-evoked response as measured by optical coherence tomography (OCT). Methods C57BL/6J mice were maintained in the dark for ∼20 hours and studied by OCT before and after 1 hour of light exposure. OR thickness in dark or light was measured, and the light-dark difference (i.e., the photoresponse) was calculated. Subgroups of mice were given either saline or d-cis-diltiazem (an inducer of transient and nondamaging OR oxidative stress) ± methylene blue (24 hours before examination) and α-lipoic acid (1 hour before examination); one group was kept only in the dark and given only AOs. Results In uninjected or saline-injected control mice, the OR showed a similar and reproducible light-induced expansion; dark-adapted mice given AOs did not increase dark-adapted OR thickness. The d-cis-diltiazem–treated mice had no photoresponse (P > 0.05). The d-cis-diltiazem–treated mice given AOs corrected (P < 0.05) the suppressed OR photoresponse, indicating the presence of oxidative stress. Conclusions QUEnch-assiSTed (QUEST) OCT reproduced results from previous gold standard assays, showing that oxidative stress impairs the OR photoresponse and that d-cis-diltiazem produces OR oxidative stress. We envision future applications of QUEST OCT in a range of oxidative stress–based retinopathies.

Purpose The rhodopsin mutation P23H is responsible for a significant portion of autosomal-dominant retinitis pigmentosa, a disorder characterized by rod photoreceptor death. The mechanisms of toxicity remain unclear; previous studies implicate destabilization of P23H rhodopsin during light exposure, causing decreased endoplasmic reticulum (ER) exit and ER stress responses. Here, we probed phototransduction in Xenopus laevis rods expressing bovine P23H rhodopsin, in which retinal degeneration is inducible by light exposure, in order to examine early physiological changes that occur during retinal degeneration. Methods We recorded single-cell and whole-retina responses to light stimuli using electrophysiology. Moreover, we monitored morphologic changes in rods after different periods of light exposure. Results Initially, P23H rods had almost normal photoresponses, but following a brief light exposure varying from 4 to 32 photoisomerizations per disc, photoresponses became irreversibly prolonged. In intact retinas, rods began to shed OS fragments after a rod-saturating exposure of 12 minutes, corresponding to approximately 10 to 100 times more photoisomerizations. Conclusions Our results indicate that in P23H rods light-induced degeneration occurs in at least two stages, the first involving impairment of phototransduction and the second involving initiation of morphologic changes.

Purpose To define the nature and extent of cone photoreceptor abnormalities in diabetic individuals who have mild or no retinopathy by assessing the activation phase of cone phototransduction and the flicker ERG in these individuals. Methods Light-adapted single-flash and flicker ERGs were recorded from 20 diabetic individuals who have no clinically apparent retinopathy (NDR), 20 diabetic individuals who have mild nonproliferative diabetic retinopathy (NPDR), and 20 nondiabetic, age-equivalent controls. A-waves elicited by flashes of different retinal illuminance were fit with a delayed Gaussian model to derive Rmp3 (maximum amplitude of the massed photoreceptor response) and S (phototransduction sensitivity). Fundamental amplitude and phase of ERGs elicited by full-field sinusoidal flicker were obtained across a frequency range of 6 to 100 Hz. Results ANVOA indicated that both diabetic groups had significant S losses compared with the controls, whereas mean Rmp3 did not differ significantly among the groups. ANOVA also indicated significantly reduced flicker ERG amplitude for frequencies ≥56 Hz for both diabetic groups compared with the controls. Flicker ERG timing (phase) did not differ significantly among the groups. Log Rmp3 + log S was significantly correlated with the patients' high-frequency (62.5 Hz) flicker ERG amplitude loss (r = 0.69, P < 0.001). Conclusions The delayed Gaussian a-wave model is useful for characterizing abnormalities in the activation phase of cone phototransduction and can help explain flicker ERG abnormalities in early-stage diabetic retinopathy. Reduced cone sensitivity and attenuated high-frequency flicker ERGs provide evidence for impaired cone function in these individuals.

Purpose: A local colony of inbred mice (129S6/SvEvTac origin), in isolation for over a decade, were found to have absent light-adapted electroretinogram (ERG) responses. We investigated the inheritance and genetic basis of this phenotype of cone photoreceptor function loss. Methods: An affected 129S6/SvEvTac colony animal was outcrossed to a C57BL/6J mouse and intercrossed to investigate inheritance in the F2 generation. We performed ERG testing and targeted resequencing on genes of interest (Gnat2, Cnga3, Cngb3, Pde6c, Hcn1, Syne2). The eyes of a subset of animals underwent histologic immunostaining. Results: All 129S6/SvEvTac colony animals tested lacked cone pathway function by ERG testing (n = 12), although rod pathway–based ERG responses remained unaffected. Outcross-intercross breeding showed a recessive inheritance pattern. A novel missense mutation was identified in the Cngb3 gene, which causes an amino acid substitution at a conserved residue (NM_013927)c.692G>A; p.(R231H). The recessive phenotype only affected homozygotes (χ2 = 39, P = 3.2e−10). Cones had normal morphology at postnatal day (PND) 70, but cone cell counts declined from PND 30 to PND 335 (P = 0.038), indicating progressive cone photoreceptor death. Conclusions: We identified the spontaneous occurrence of a 10th model of cone photoreceptor function loss (cpfl10) in an isolated line of inbred mice. Our results indicate that this is caused by a novel missense mutation in the Cngb3 gene, with a fully recessive inheritance pattern. This mouse may provide a more appropriate background against which to assess CNGB3 achromatopsia gene therapy for missense mutations.

Purpose To describe the retinal phenotype of pediatric patients with mutations in the retinol dehydrogenase 12 (RDH12) gene. Methods Twenty-one patients from 14 families (ages 2-17 years) with RDH12-associated inherited retinal degeneration (RDH12-IRD) underwent a complete ophthalmic exam and imaging with spectral domain optical coherence tomography (SD-OCT) and near infrared and short-wavelength fundus autofluorescence. Visual field extent was measured with Goldmann kinetic perimetry, visual thresholds with dark-adapted static perimetry or with dark-adapted chromatic full-field stimulus testing (FST) and transient pupillometry. Results Visual acuity ranged from 20/40 to light perception. There was parafoveal depigmentation or atrophic maculopathies accompanied by midperipheral intraretinal pigment migration. SD-OCT revealed foveal thinning in all patients and detectable but thinned outer nuclear layer (ONL) at greater eccentricities from the fovea. Photoreceptor outer segment (POS) signals were only detectable in small pockets within the central retina. Measurable kinetic visual fields were limited to small (

Purpose To determine the progression rate and the variability of rod and cone sensitivities in patients with X-linked retinitis pigmentosa (XLRP) caused by mutations in ORF15-RPGR. Methods ORF15-RPGR-XLRP patients (n = 15) were studied prospectively over 2 years with static perimetry sampling the visual field under dark-adapted and light-adapted conditions on a 12° square grid covering 168° width and 84° height. Natural history of rod and cone sensitivity loss and test-retest variability were estimated. Data were analyzed pointwise as well as averaged across small regions of neighboring loci of approximately 80 mm2 (900 deg2) in size representing the likely extent of localized gene therapy injections. Results Retinal loci with mild to moderate loss of sensitivity tended to be in the mid- to far-peripheral retina in most patients. When averaged across small regions, dark-adapted rod vision progressed at an average of 2 dB per year with a coefficient of repeatability (CR) of 6.3 dB, and light-adapted cone vision with white stimulus progressed at an average of 0.9 dB per year with a CR of 3.8 dB. For an average patient enrolled in an early-phase clinical trial, significant (α = 0.05) progression would be predicted to occur with 80% power in 4.5 years for rod vision and 6.1 years for cone vision. Localization of regions in the temporal hemifield and grouping of results from multiple patients would permit trial designs of shorter duration. Conclusions Measurement of rod sensitivity under dark-adapted conditions averaged across a small region showed the greatest potential for detectability of progression in the shortest period.

Purpose There is ample evidence that retinal dopamine (DA) is involved in the biochemical signaling cascade that controls emmetropization, but it is unknown how its release depends on the spectral composition of ambient light. We have studied DA release, refractive development, and growth in chicken eyes that were exposed to light of different spectral bands, and had either normal vision or were covered by frosted diffusers to induce myopia. Methods Experiment 1: After spending the night in the dark, chicks were exposed to white room light (spectral range, 430-630 nm) or kept in the dark. Additional chicks were unilaterally exposed to blue (peak at 470 nm), red (620 nm), or UV lighting (375 nm) for 30 minutes and their fellow eyes covered with black occluders to minimize light exposure. Experiment 2: In the second experiment, chicks wore diffusers over one eye to induce deprivation myopia and were raised for 5 days in either white room light or in lighting supplied by UV, blue, or red light-emitting diodes (LEDs). Refractive states were recorded daily with infrared photoretinoscopy, and ocular dimensions at the start and end of the experiment with A-scan ultrasonography. DA and its metabolites were measured in retina and vitreous by high pressure liquid chromatography-electrochemical detection (HPLC-ED) in all cases. Results Compared to chicks kept in the dark, retinal DA and vitreal 3,4-dihydroxyphenylacetic acid (DOPAC) concentrations were clearly elevated after 30 minutes in white light. Vitreal DOPAC was also increased in red, blue, and UV lighting, compared to the fellow eyes covered with black occluders (black occluder versus blue light: 1.31 ± 0.32 vs. 1.70 ± 0.37; red: 1.26 ± 0.33 vs. 1.64 ± 0.38; UV: 1.13 ± 0.19 vs. 1.63 ± 0.21 ng/0.1 g wet weight). Chickens developed significantly less deprivation myopia, with shorter eyes, when raised under UV and blue lighting for 5 days, compared to under red and white light. Eyes with normal vision became more hyperopic in blue and UV lighting. Vitreal DOPAC levels were lowest after 5 days of exposure to UV lighting. Conclusions Red, blue, and UV lighting all stimulated the release of retinal DA, but there were wavelength-dependent differences in DA release and metabolism. Less deprivation myopia developed in UV and blue lighting, compared to white and red light. The application of these findings to humans is limited by the fact that, different from chicks, humans have very low sensitivity in the near-UV region of the spectrum.

Purpose: To compare static rod function obtained with and without photobleach in control and intermediate age-related macular degeneration (iAMD) participants with and without subretinal drusenoid deposits (SDD). Methods: In this cross-sectional study, retinal sensitivities within the central 24° retina were obtained twice using a dark-adapted chromatic perimeter, both with 505- and 625-nm stimuli. Tests were performed after 30 minutes of dark-adaptation either with or without a preceding photobleach. Multimodal imaging was performed to grade AMD and SDD status, and other retinal changes considered being risk factors for progression to late AMD. The sensitivity difference between both stimuli was used to assess rod function. The average point wise sensitivity difference (PWSD) was compared among the study groups. Results: Twenty-nine control subjects and 20 iAMD without SDD and 17 iAMD with SDD cases were recruited. The average PWSD of the SDD group was significantly reduced (more with photobleach) compared with that of the control (P < 0.001) and no-SDD groups (P < 0.001), but only within the central 8°. The average PWSD of the non-SDD group was also reduced compared with the control group but only for measurements with photobleach (P = 0.020). There was no difference in average PWSD between the presence and absence of hyperreflective foci and/or nascent geographic atrophy in iAMD eyes without SDD (P = 0.60) or with SDD (P = 0.12). Conclusions: iAMD eyes with SDD are associated with worse static rod function compared with eyes without SDD. The greatest abnormality in rods is observed within the central 8° and when tested with a preceding photobleach.

Purpose: The primary aim was to determine optimal test conditions for evaluating dark adaptation in intermediate age-related macular degeneration (iAMD) in order to minimize test time while maintaining diagnostic sensitivity.\ud \ud Methods: People with AMD and age-similar controls were recruited (aged >55 years). Rod intercept time (RIT) was assessed after a 76%, 70%, and 65% rhodopsin bleach at 5° eccentricity and 76% and 70% bleach at 12°. Test order was randomized and a 30-minute washout period added between tests. Results were compared between control and iAMD groups and receiver operating characteristics (ROC) curves were constructed.\ud \ud Results: A total of 26 participants with variable grades of macular health attended for two visits. There was a statistically significant difference in average RIT between the control and iAMD groups at 5° (median, IQR controls = 5.8 minutes, 3.8–7.5; iAMD = 20.6 minutes, 11.1–30.0; Mann-Whitney, P = 0.01) and at 12° (mean, controls: 4.54 minutes ± 2.12 SD, iAMD = 7.72 minutes ± 3.37 SD; independent samples t-test, P = 0.03) following a 76% bleach. Area under the ROC curves was 0.83 (confidence interval [CI]: 0.64–1.0) and 0.79 (CI: 0.59–0.99) for these two test conditions, respectively. Five participants (45%) in the iAMD group had RITs >20 minutes for 76% bleach at 5°, but none for any other test condition.\ud \ud Conclusions: Nearly half of the participants with iAMD produced unacceptably long recovery times (>20 minutes) using a 76% bleach at 5° eccentricity. The 76% bleach at 12° provided almost equivalent separation between AMD and controls but recovery was achieved within 20 minutes.

Purpose To examine spatial changes in rod-mediated function in relationship to local structural changes across the central retina in eyes with a spectrum of age-related macular degeneration (AMD) disease severity. Methods Participants were categorized into five AMD severity groups based on fundus features. Scotopic thresholds were measured at 14 loci spanning ±18° along the vertical meridian from one eye of each of 42 participants (mean = 71.7 ± 9.9 years). Following a 30% bleach, dark adaptation was measured at eight loci (±12°). Rod intercept time (RIT) was defined from the time to detect a −3.1 log cd/m2 stimulus. RITslope was defined from the linear fit of RIT with decreasing retinal eccentricity. The presence of subretinal drusenoid deposits (SDD), ellipsoid (EZ) band disruption, and drusen at the test loci was evaluated using optical coherence tomography. Results Scotopic thresholds indicated greater rod function loss in the macula, which correlated with increasing AMD group severity. RITslope, which captures the spatial change in the rate of dark adaptation, increased with AMD severity (P < 0.0001). Three rod function phenotypes emerged: RF1, normal rod function; RF2, normal scotopic thresholds but slowed dark adaptation; and RF3, elevated scotopic thresholds with slowed dark adaptation. Dark adaptation was slowed at all loci with SDD or EZ band disruption, and at 32% of loci with no local structural changes. Conclusions Three rod function phenotypes were defined from combined measurement of scotopic threshold and dark adaptation. Spatial changes in dark adaptation across the macula were captured with RITslope, which may be a useful outcome measure for functional studies of AMD.

Purpose Using optical coherence tomography (OCT) to analyze the effects of light/dark adaptation in a mouse model of inherited photoreceptor degeneration (rd10), and to study dynamics of subretinal fluid during the progress of retinal degeneration. Methods rd10 and wild-type (WT) C57BL/6J mice were reared in cyclic light or darkness and imaged with Bioptigen UHR-OCT or Spectralis HRA+OCT after adaptation to either light or darkness. Results OCT images from rd10 mice were analyzed at three progressive stages of degeneration. After light-adaptation, stage I (postnatal age [P]26-29) eyes demonstrated no apparent subretinal fluid. At stage II (P32-38), subretinal fluid was present and restricted to parapapillary area, while at stage III (P44-45) extensive subretinal fluid was present across many retinal areas. Following overnight dark-adaptation, WT eyes showed a large reduction in outer retinal thickness (4.6 ± 1.4 μm, n = 16), whereas this change was significantly smaller in stage I rd10 eyes (1.5 ± 0.5 μm, n = 14). In stage II rd10 eyes, dark-adaptation significantly reduced the extent of subretinal fluid, with the amount of reduction correlating with the amount of fluid pre-existing in the light-adapted state. However, dark-adaptation did not significantly alter the amount of subretinal fluid observed in stage III rd10 mice. In addition, dark-rearing of rd10 mice from P6 to P30 slowed retinal degeneration. Conclusions Visual experience in the form of light/dark adaptation exerts a significant effect on outer retinal structure in the context of photoreceptor degeneration. This effect may arise from light-dependent alterations in fluid transport across the RPE monolayer, and promote photoreceptor survival as induced by dark-rearing.

Sensitivity regulation enables the visual system to function effectively from the absorption of a few photons at the lowest visual threshold to the absorption of enough photons to bleach nearly all the light-sensitive photopigment in the eye. Here, we investigate sensitivity regulation in the rod (or scotopic) range from -3.8 to -0.8 log(10) scotopic trolands. Over most of this range, the rate of photon absorption per rod is too low for sensitivity regulation to be practicable within the rod photoreceptor itself, so that regulation must occur postreceptorally. We measured adaptation-dependent changes in visual sensitivity and visual delay, which together provide a much more complete characterization of the effects of light adaptation than the usual method of measuring sensitivity changes alone. Our results demonstrate clearly that changes in scotopic sensitivity with increasing light levels are achieved in large part by a speeding up of the scotopic response and a decrease in the temporal integration time. Thus, the scotopic and the photopic systems both regulate their steady-state sensitivity using the same strategy, even though the scotopic system does it largely postreceptorally and the photopic system largely receptorally.

Purpose To evaluate the effect of acutely elevated intraocular pressure (IOP) on the functional and blood flow responses of the rat retina to flicker stimulation. Methods Brown Norway (n = 15) rats were dark-adapted before ketamine/xylazine anesthesia. IOP was raised acutely in one eye to ∼45 mm Hg with a vascular loop. In 11 rats, white light flicker stimulus (10 Hz, 2 seconds duration, 0.80 log scotopic cd·s/m2) was applied before and during IOP elevation, and 10 minutes after loop removal. Changes in the total retinal blood flow (TRBF) and retinal function induced by the visual stimulus were measured simultaneously with a combined optical coherence tomography (OCT) + electroretinography (ERG) system. Systemic blood pressure was measured in the remaining four rats frequently from 10 to 90 minutes post anesthesia injection. Results The systemic blood pressure remained at 99 ± 4 mm Hg throughout the measurements (n = 4). Under normal IOP, the TRBF was 5.6 ± 1.9 μL/min, and the average retinal blood vessel size (BVS) in the vicinity of the optic nerve head (ONH) was 44.1 ± 4.5 μm. During IOP elevation, the TRBF was significantly lower (3.8 ± 1.2 μL/min, P < 0.01) and the BVS was significantly smaller (35.1 ± 2.6 μm, P < 0.01). Both TRBF and BVS returned to baseline within ∼10 minutes from removal of the vascular loop. The flicker-induced TRBF change measured under normal IOP (6.0 ± 3.3%) was reduced significantly to 0.1 ± 0.3% (P < 0.01) during IOP elevation, and recovered to 5.9 ± 1.7% within 10 minutes after loop removal. During IOP elevation, the magnitude of the ERG second harmonic component (SHC) decreased to 55% of its baseline value (P < 0.01) and remained significantly smaller than baseline (P < 0.01). Conclusions Acute IOP elevation to 45 mm Hg caused suppression of the retinal functional and TRBF response to flicker stimulation.

Purpose To characterize the natural history of rod-mediated dark adaptation (RMDA) over 2 years in eyes with intermediate age-related macular degeneration (AMD). This information will be useful in understanding the potential of RMDA to serve as a functional endpoint in proof-of-concept studies and clinical trials on intermediate AMD. Methods RMDA was measured in eyes with intermediate AMD at baseline and follow-up visits over 2 years at 6, 12, 18, and 24 months. A computerized dark adaptometer measured sensitivity for targets centered at 11° on the superior vertical meridian of the retina. Rod intercept time (RIT) characterized the speed of dark adaptation and was defined as the duration (in minutes) required for sensitivity to reach a criterion level of 3.0 log units of attenuation of the stimulus. Results Mean change in RIT over 24 months for 30 eyes was 10.5 minutes (standard deviation 19.4), p < 0.0001; 73.3% of eyes had a RIT increase >1 minute, 56.7% had an increase >3 minutes, and 36.7% had an increase >6 minutes; for 26.7% RIT was unchanged (0- to 1-minute increase) or decreased. Greater increase in RIT over 24 months was associated with smoking. Conclusions RMDA slows in intermediate AMD over 2 years in most eyes. There was wide variability in RIT at both baseline and in the extent to which it increased over 24 months. A major risk factor for AMD, smoking, exacerbated RMDA slowing. Translational Relevance RMDA as assessed by RIT may be useful as a functional endpoint in proof-of-concept studies and clinical trials on intermediate AMD with 2-year designs.

Purpose To investigate the chromatic characteristics and intensity-response function of light-induced reflex lacrimation and its correlation with the melanopsin-driven postillumination pupil response (PIPR). Methods Eleven visually normal participants completed the experiment. Lacrimation was measured in one eye by placing a calibrated filter paper strip in the conjunctival sac over a 1 minute-interval (Schirmer's test) during which participants received either no light stimulation (baseline trial) or one flash of blue or red light stimuli presented binocularly with a Ganzfeld stimulator, while the pupil response was recorded simultaneously from the fellow eye by using an eye tracker. Light stimulation trials were presented in alternating fashion at seven incremental intensity steps (0.1, 1, 3.16, 10, 31.6, 100, and 400 cd/m2). Postillumination pupil response was defined as the mean pupil constriction from 10 to 30 seconds post illumination. Results The amount of lacrimation in response to 10 to 400 cd/m2 blue light was significantly greater than baseline and increased monotonically with increasing light intensity. Red light did not induce significant reflex lacrimation until the brightest stimulation at 400 cd/m2. There was a positive linear correlation between PIPR and lacrimation in response to blue light (r = 0.74, P < 0.001) but not to red light (r = 0.13, P = 0.25). Conclusions The chromatic characteristics and intensity-response of light-induced lacrimation are highly consistent with the features of melanopsin phototransduction. This finding is the first in vivo evidence in humans, supporting the hypothesis that light-induced reflex lacrimation is mediated primarily by melanopsin photoactivity, and provides new insight into the putative mechanisms of photophobia.

Purpose Two-photon excited fluorescence (TPEF) imaging has potential as a functional tool for tracking visual pigment regeneration in the living eye. Previous studies have shown that all-trans-retinol is likely the chief source of time-varying TPEF from photoreceptors. Endogenous TPEF from retinol could provide the specificity desired for tracking the visual cycle. However, in vivo characterization of native retinol kinetics is complicated by visual stimulation from the imaging beam. We have developed an imaging scheme for overcoming these challenges and monitored the formation and clearance of retinol. Methods Three macaques were imaged by using an in vivo two-photon ophthalmoscope. Endogenous TPEF was excited at 730 nm and recorded through the eye's pupil for more than 90 seconds. Two-photon excited fluorescence increased with onset of light and plateaued within 40 seconds, at which point, brief incremental stimuli were delivered at 561 nm. The responses of rods to stimulation were analyzed by using first-order kinetics. Results Two-photon excited fluorescence resulting from retinol production corresponded to the fraction of rhodopsin bleached. The photosensitivity of rhodopsin was estimated to be 6.88 ± 5.50 log scotopic troland. The rate of retinol clearance depended on intensity of incremental stimulation. Clearance was faster for stronger stimuli and time constants ranged from 50 to 300 seconds. Conclusions This study demonstrates a method for rapidly measuring the rate of clearance of retinol in vivo. Moreover, TPEF generated due to retinol can be used as a measure of rhodopsin depletion, similar to densitometry. This enhances the utility of two-photon ophthalmoscopy as a technique for evaluating the visual cycle in the living eye.

Purpose Mutations in the retinal transcription factor cone-rod homeobox (CRX) gene result in severe dominant retinopathies. A large animal model, the Rdy cat, carrying a spontaneous frameshift mutation in Crx, was reported previously. The present study aimed to further understand pathogenesis in this model by thoroughly characterizing the Rdy retina. Methods Structural and functional changes were found in a comparison between the retinas of CrxRdy/+ kittens and those of wild-type littermates and were determined at various ages by fundus examination, electroretinography (ERG), optical coherence tomography, and histologic analyses. RNA and protein expression changes of Crx and key target genes were analyzed using quantitative reverse-transcribed PCR, Western blot analysis, and immunohistochemistry. Transcription activity of the mutant Crx was measured by a dual-luciferase transactivation assay. Results CrxRdy/+ kittens had no recordable cone ERGs. Rod responses were delayed in development and markedly reduced at young ages and lost by 20 weeks. Photoreceptor outer segment development was incomplete and was followed by progressive outer retinal thinning starting in the cone-rich area centralis. Expression of cone and rod Crx target genes was significantly down-regulated. The mutant Crx allele was overexpressed, leading to high levels of the mutant protein lacking transactivation activity. Conclusions The CrxRdy mutation exerts a dominant negative effect on wild-type Crx by overexpressing mutant protein. These findings, consistent with those of studies in a mouse model, support a conserved pathogenic mechanism for CRX frameshift mutations. The similarities between the feline eye and the human eye with the presence of a central region of high cone density makes the CrxRdy/+ cat a valuable model for preclinical testing of therapies for dominant CRX diseases.

Night myopia, which is a shift in refraction with light level, has been widely studied but still lacks a complete understanding. We used a new infrared open-view binocular Hartmann-Shack wave front sensor to quantify night myopia under monocular and natural binocular viewing conditions. Both eyes' accommodative response, aberrations, pupil diameter, and convergence were simultaneously measured at light levels ranging from photopic to scotopic conditions to total darkness. For monocular vision, reducing the stimulus luminance resulted in a progression of the accommodative state that tends toward the subject's dark focus or tonic accommodation and a change in convergence following the induced accommodative error. Most subjects presented a myopic shift of accommodation that was mitigated in binocular vision. The impact of spherical aberration on the focus shift was relatively small. Our results in monocular conditions support the hypothesis that night myopia has an accommodative origin as the eye progressively changes its accommodation state with decreasing luminance toward its resting state in total darkness. On the other hand, binocularity restrains night myopia, possibly by using fusional convergence as an additional accommodative cue, thus reducing the potential impact of night myopia on vision at low light levels.

Purpose The purpose of this study was to develop a convenient means to measure rod (and cone) function by automated perimetry in patients with inherited retinal degenerations (IRDs). Methods A currently available automated perimeter was used to determine sensitivity (in decibels) to a blue target in the dark-adapted (DA) state and a white target in the light-adapted (LA) state. Normal subjects and IRD patients were evaluated with a full-threshold 71-locus strategy (the retinitis pigmentosa [RP] test) and a size III target. Comparisons were made with results from the more commonly used methods of two-color DA perimetry and middle/long-wavelength LA perimetry in the same patients. Results Rod function using the blue target and the RP test was determined for normal subjects by measuring DA sensitivities. If patients detected the blue stimulus in the DA state, it was determined whether the value was rod mediated by using normal data acquired during the cone plateau phase of dark adaptation. If rod mediated, rod sensitivity loss (RSL) was calculated and mapped across the visual field. Light-adapted sensitivities in normal subjects were also measured, permitting cone sensitivity losses (CSL) to be calculated for the patients. Multiple methods were used to compare RSL and CSL results with those from two-color DA perimetry and chromatic LA perimetry, and there was close correspondence between the methods. Conclusions The unmodified automated static perimeter used in the DA and LA states presents a practical approach to accomplish current goals of treatment trials in IRDs. This proof-of-principle study is an initial step toward establishing a clinical method to gather reproducible data on photoreceptor-mediated sensitivity.

Purpose To compare the impact of intraocular pressure (IOP) elevation on scotopic and photopic contrast sensitivity in mice. Methods We chronically elevated the IOP of wild-type mice via injection of polystyrene beads or acutely via injection of highly cohesive sodium hyaluronate. Some eyes with chronically elevated IOP were treated with either topical brimonidine tartrate 0.1% or brinzolamide 1%. Scotopic and photopic contrast sensitivity was assessed at peak spatiotemporal frequencies at multiple time points, with an established optokinetic technique. Retinal ganglion cell (RGC) counts were determined with an antibody to class III beta-tubulin. Correlations among IOP level, RGC count, and scotopic or photopic contrast sensitivity were performed. Results Six weeks of IOP elevation caused a generalized reduction of photopic contrast sensitivity and a preferential reduction of scotopic contrast sensitivity at peak spatiotemporal frequencies. The administration of brinzolamide but not brimonidine caused a significant reduction in cumulative IOP, whereas brimonidine, but not brinzolamide, prevented RGC loss. Both brimonidine and brinzolamide prevented contrast sensitivity loss, but brimonidine did so at earlier time points and across a wider range of lighting conditions. Following either chronic or acute IOP elevation, scotopic contrast sensitivity was impacted most prominently by IOP level and not by RGC count, while photopic contrast sensitivity was impacted by a combination of factors. Conclusions It is possible that scotopic-specific retinal circuitry is altered preferentially by IOP elevation, and that changes in scotopic contrast sensitivity will assist with glaucoma detection. Brimonidine appears to prevent RGC loss via an IOP-independent mechanism.

Purpose Moderately elevated intraocular pressure (IOP) is a risk factor for open-angle glaucoma. Some patients suffer glaucoma despite clinically measured normal IOPs. Fluctuations in IOP may have a significant role since IOPs are higher during sleep and inversion activities. Controlled transient elevations of IOPs in rats over time lead to optic nerve structural changes that are similar to the early changes observed in constant chronic models of glaucoma. Because early intervention decreases glaucoma progression, this study was done to determine if early physiological changes to the retina could be detected with noninvasive electrophysiological and optical imaging tests during moderately elevated IOP. Methods Intraocular pressures were raised to moderately high levels (35 mm Hg) in one eye of Sprague-Dawley rats while the other (control) eye was untreated. One group of rats underwent scotopic threshold response (STR) and electroretinogram (ERG) testing, while another 3 groups underwent optical coherence tomography (OCT) imaging, Western blot, or histologic evaluation. Results The amplitudes of the STR and ERG responses in eyes with moderately elevated IOPs were enhanced compared to the values before IOP elevation, and compared to untreated contralateral eyes. Structural changes to the optic nerve also occurred during IOP elevation. Conclusions Although ischemic IOP elevations are well-known to globally reduce components of the scotopic ERG, acute elevation in rats to levels often observed in untreated glaucoma patients caused an increase in these parameters. Further exploration of these phenomena may be helpful in better understanding the mechanisms mediating early retinal changes during fluctuating or chronically elevated IOP.

Purpose Point and null mutations in interphotoreceptor retinoid-binding protein (IRBP) cause retinal dystrophy in affected patients and IRBP-deficient mice with unknown mechanism. This study investigated whether IRBP protects cells from damages induced by all-trans-retinal (atRAL), which was increased in the Irbp(-/-) retina. Methods Wild-type and Irbp(-/-) mice retinal explants in buffer with or without purified IBRP were exposed to 800 lux light for different times and subjected to retinoid analysis by high-performance liquid chromatography. Purity of IRBP was determined by Coomassie Brilliant Blue staining and immunoblot analysis. Cellular damages induced by atRAL in the presence or absence of IRBP were evaluated in the mouse photoreceptor-derived 661W cells. Cell viability and death were measured by 3-(4,5-dimethyl-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) and TUNEL assays. Expression and modification levels of retinal proteins were determined by immunoblot analysis. Intracellular reactive oxygen species (ROS) and nitric oxide (NO) were detected with fluorogenic dyes and confocal microscopy. Mitochondrial membrane potential was analyzed by using JC-1 fluorescent probe and a flow cytometer. Results Content of atRAL in Irbp(-/-) retinal explants exposed to light for 40 minutes was significantly higher than that in wild-type retinas under the same light conditions. All-trans-retinal caused increase in cell death, tumor necrosis factor activation, and Adam17 upregulation in 661W cells. NADPH oxidase-1 (NOX1) upregulation, ROS generation, NO-mediated protein S-nitrosylation, and mitochondrial dysfunction were also observed in 661W cells treated with atRAL. These cytotoxic effects were significantly attenuated in the presence of IRBP. Conclusions Interphotoreceptor retinoid-binding protein is required for preventing accumulation of retinal atRAL, which causes inflammation, oxidative stress, and mitochondrial dysfunction of the cells.

Purpose Abnormalities of blood vessel anatomy, morphology, and ratio can serve as important diagnostic markers for retinal diseases such as AMD or diabetic retinopathy. Large cohort studies demand automated and quantitative image analysis of vascular abnormalities. Therefore, we developed an analytical software tool to enable automated standardized classification of blood vessels supporting clinical reading. Methods A dataset of 61 images was collected from a total of 33 women and 8 men with a median age of 38 years. The pupils were not dilated, and images were taken after dark adaption. In contrast to current methods in which classification is based on vessel profile intensity averages, and similar to human vision, local color contrast was chosen as a discriminator to allow artery vein discrimination and arterial-venous ratio (AVR) calculation without vessel tracking. Results With 83% ± 1 standard error of the mean for our dataset, we achieved best classification for weighted lightness information from a combination of the red, green, and blue channels. Tested on an independent dataset, our method reached 89% correct classification, which, when benchmarked against conventional ophthalmologic classification, shows significantly improved classification scores. Conclusions Our study demonstrates that vessel classification based on local color contrast can cope with inter- or intraimage lightness variability and allows consistent AVR calculation. We offer an open-source implementation of this method upon request, which can be integrated into existing tool sets and applied to general diagnostic exams.

Mesopic and scotopic vision extend over an illuminance range of 106. The goal of the present study was to determine the effect of decreasing light level on the underlying motion mechanism that integrates spatiotemporally separated motion signals. To accomplish this, we took advantage of the phenomenon of visual motion priming, in which the perceived direction of a directionally ambiguous test stimulus is influenced by the directional movement of a preceding priming stimulus. After terminating a drifting priming stimulus, a 180° phase-shifted grating was presented as a test stimulus. The priming and test stimuli were separately presented to the central and peripheral retinas, respectively. The participants judged the perceived direction of this test stimulus at various light levels from photopic to scotopic levels. We found that the effects of motion priming disappeared over 1 log unit of mesopic light levels. When the test stimulus was presented before the offset of the priming stimulus to compensate for the temporal delay in the rod pathway or when both stimuli were presented at the same location in the periphery, a motion-priming effect appeared at mesopic light levels. These results suggest that different temporal characteristics between the cone pathway and rod pathway disturb the function of the putative motion mechanism responsible for the spatiotemporal integration of motion signals, which leads to specific modulation of motion perception over a wide range of mesopic vision.

Purpose The pathophysiology of vision loss in persons with diabetic retinopathy (DR) is complex and incompletely defined. We hypothesized that retinal pigment epithelium (RPE) and rod and cone photoreceptor dysfunction, as measured by dark adaptometry, would increase with severity of DR, and that pan-retinal photocoagulation (PRP) would exacerbate this dysfunction. Methods Dark adaptation (DA) was measured in subjects with diabetes mellitus and healthy controls. Dark adaptation was measured at 5° superior to the fovea following a flash bleach, and the data were analyzed to yield cone and rod sensitivity curves. Retinal layer thicknesses were quantified using spectral-domain optical coherence tomography (OCT). Results The sample consisted of 23 controls and 73 diabetic subjects. Subjects with moderate nonproliferative diabetic retinopathy (NPDR) exhibited significant impairment of rod recovery rate compared with control subjects (P = 0.04). Cone sensitivity was impaired in subjects with proliferative diabetic retinopathy (PDR) (type 1 diabetes mellitus [T1DM]: P = 0.0047; type 2 diabetes mellitus [T2DM]: P < 0.001). Subjects with untreated PDR compared with subjects treated with PRP exhibited similar rod recovery rates and cone sensitivities. Thinner RPE as assessed by OCT was associated with slower rod recovery and lower cone sensitivity, and thinner photoreceptor inner segment/outer segment layer was associated with lower cone sensitivity. Conclusions The results suggest that RPE and photoreceptor cell dysfunction, as assessed by cone sensitivity level and rod- and RPE-mediated dark adaptation, progresses with worsening DR, and rod recovery dysfunction occurs earlier than cone dysfunction. Function was preserved following PRP. The findings suggest multiple defects in retinoid function and provide potential points to improve visual function in persons with PDR.