Melanopsin-mediated amplification of cone signals in the human visual cortex.

The ambient daylight variation is coded by melanopsin photoreceptors and their luxotonic activity increases towards midday when colour temperatures are cooler, and irradiances are higher. Although melanopsin and cone photoresponses can be mediated via separate pathways, the connectivity of melanopsin cells across all levels of the retina enables them to modify cone signals. The downstream effects of melanopsin-cone interactions on human vision are however, incompletely understood. Here, we determined how the change in daytime melanopsin activation affects the human cone pathway signals in the visual cortex. A 5-primary silent-substitution method was developed to evaluate the dependence of cone-mediated signals on melanopsin activation by spectrally tuning the lights and stabilizing the rhodopsin activation under a constant cone photometric luminance. The retinal (white noise electroretinogram) and cortical responses (visual evoked potential) were simultaneously recorded with the photoreceptor-directed lights in 10 observers. By increasing the melanopsin activation, a reverse response pattern was observed with cone signals being supressed in the retina by 27% (p = 0.03) and subsequently amplified by 16% (p = 0.01) as they reach the cortex. We infer that melanopsin activity can amplify cone signals at sites distal to retinal bipolar cells to cause a decrease in the psychophysical Weber fraction for cone vision.

The role of melanopsin photoreception on visual attention linked pupil responses.

A decision during a visual task is marked by a task-evoked pupil dilation (TEPD) that is linked to the global cortical arousal state. Melanopsin expressing intrinsically photosensitive retinal ganglion cells (ipRGCs) form the afferent pathway for this pupil response. Melanopsin activation also influences mood and arousal and increases activity in decision-making brain areas that receive direct ipRGC projections. Here, an optical photostimulation method controlled the excitations of all five photoreceptor classes in the human eye to isolate melanopsin-mediated photoreception. We hypothesised that the TEPD can be driven by directing active visual covert attention through the ipRGC pathway. When observers are completely certain of the stimulus presence, melanopsin-directed stimulation produces a TEPD of similar amplitude to a cone-directed stimulation, with their combination producing larger amplitudes. This dilation is satisfactorily modelled by linear addition with a higher melanopsin weighting in ipRGCs. Visual reaction times were longest in response to melanopsin-directed lights. Next, we asked whether the afferent photoreceptor input and decision certainty, controlled by priming the observer's a priori expectation, interact to drive the TEPD. Signal detection analysis showed that by fixing the predecision certainty (bias), the phasic arousal and TEPD amplitude vary with observer criterion (c') and sensitivity (d') but not with preferential activation of melanopsin. The signature feature of the melanopsin response during attention was a biphasic TEPD. We conclude that active covert attention can be modulated by visual information mediated via ipRGCs, but that phasic arousal responses marked using the TEPD are not increased by higher levels of melanopsin activation.

Design and validation of a chart-based measure of the limits of spatial contrast sensitivity.

PURPOSE: Current chart-based tests of spatial contrast sensitivity (SCS) with fixed or narrow frequency ranges (≤18 cycles/°) cannot characterise the limits of spatial contrast vision. Here we present the design and validation of a chart-based measure of the spatial contrast envelope. METHODS: Following the principles of the standard visual acuity (Bailey-Lovie) and contrast sensitivity (Pelli-Robson) charts, a combined spatial-contrast and visual acuity chart was designed using a language-independent triangular symbol for a four-alternative forced-choice procedure plus chart rotation. Symbol frequencies ranged between 0.38 and 60 cycles/° spaced along 10 radial axes (0.55%-100% contrast). The chart was validated with reference to the Bailey-Lovie and Pelli-Robson charts; its reliability and sensitivity to changes in illumination, simulated cataract and blur was evaluated in healthy adults. RESULTS: The photopic SCS function could be measured in 5.5 ± 0.5 min; thresholding around the spatial contrast resolution limit reduced completion times to ~2 min. There was good agreement with high-contrast visual acuity (difference = 0.08 ± 0.02 logMAR) and contrast-sensitivity at 1.5 cycles/° (0.13 ± 0.06 logCS). Test-retest reliability was excellent at all spatial frequencies (ICC = 0.99). Mesopic illumination or simulated cataract caused a generalised SCS loss; myopic blur reduced high-frequency sensitivity. Spatial contrast sensitivity was independent of radial axis orientation (cardinal or oblique). CONCLUSIONS: The chart provides a time-efficient, reliable and inexpensive measure of SCS with applications in research and clinic for detecting subtle deficits in early stages of ocular and neurological conditions that often manifest at higher frequencies. It is sensitive to vision changes occurring in dim lighting and with simulated cataract and blur. The chart is available open-access for self-printing; contrast variation in print can be controlled through user calibration and/or establishing normative SCS functions using the theoretical values.

A five-primary Maxwellian-view display for independent control of melanopsin, rhodopsin, and three-cone opsins on a fine spatial scale.

Independent spatiotemporal control of the stimulation of the five photoreceptor classes requires a display with as many primary lights to probe their isolated spatial and temporal responses. No such system exists with suitable performance properties. We present a system to construct a five-primary display from commercially available three-primary digital light processing projectors. It optimizes the set of five primary lights required to maximize the achievable contrast of a single photoreceptor class in a silent substitution protocol, including where the background chromaticity is first specified. From these chosen five primaries, we describe a method to convert complex three-primary (RGB) images to five-primary representations with complete specification of the photoreceptor excitations at each pixel. Key to enabling this multiple display system with a single HDMI connection is a novel control protocol implemented in a deterministic field-programmable gate array controller that splits the data into five video streams to allow nearly synchronous presentation of primary image data through multiple displays. Each pixel is controlled over 9.5 bits for each primary over a single frame for measurement of threshold-level vision. In addition to a large contrast gamut, the Maxwellian view offers high retinal illumination to support the investigation of five opsin-based responses to complex spatiotemporal images with a truly silent substitution protocol, while avoiding the confounding effects of uncontrolled photoreceptor excitations as occurs in four-primary systems. The customizable primaries facilitate this display translation to species with different photoreceptor spectral responses, and the optics are designed for integration into microscopes for use as a stimulus generator in physiological experiments.

Melanopsin hypersensitivity dominates interictal photophobia in migraine.

PURPOSE: To define the melanopsin and cone luminance retinogeniculate pathway contributions to photophobia in healthy controls and migraineurs. METHODS: Healthy controls and migraineurs were categorized according to the International Classification of Headache Disorders criteria. Photophobia was measured under full-field illumination using electromyography in response to narrowband lights spanning the melanopsin and cone luminance action spectra. Migraineurs were tested during their interictal headache-free period. Melanopsin-mediated post-illumination pupil responses quantified intrinsically photosensitive Retinal Ganglion Cell (ipRGC) function. RESULTS: A model combining the melanopsin and cone luminance action spectra best described photophobia thresholds in controls and migraineurs; melanopsin contributions were ∼1.5× greater than cone luminance. In the illumination range causing photophobia, migraineurs had lower photophobia thresholds (∼0.55 log units; p < 0.001) and higher post-illumination pupil response amplitudes (p = 0.03) than controls. CONCLUSION: Photophobia is driven by melanopsin and cone luminance inputs to the cortex via the retino-thalamocortical pathway. In migraineurs, lower photophobia thresholds reflect hypersensitivity of ipRGC and cone luminance pathways, with the larger and prolonged post-illumination pupil response amplitude indicative of a supranormal melanopsin response. Our findings inform artificial lighting strategies incorporating luminaires with low melanopsin excitation and photopic luminance to limit the lighting conditions leading to photophobia.

Optimizing methods to isolate melanopsin-directed responses.

The intrinsic melanopsin photoresponse may initiate visual signals that differ in spatiotemporal characteristics from the cone-opsin- and rhodopsin-mediated signals. Applying the CIE standard observer functions in silent-substitution methods can require individual differences in photoreceptor spectral sensitivities and pre-receptoral filtering to be corrected; failure to do so can lead to the intrusion of more sensitive cone processes with putative melanopsin-directed stimuli. Here we evaluate heterochromatic flicker photometry (HFP) and photoreceptor-directed temporal white noise as techniques to limit the effect of these individual differences. Individualized luminous efficiency functions (V(λ)) were compared to the CIE standard observer functions. We show that adapting chromaticities used in silent-substitution methods can deviate by up to 54% in luminance when estimated with the individual and standard observer functions. These deviations lead to inadvertent cone intrusions in the visual functions measured with melanopsin-directed stimuli. To eliminate the intrusions, individual HFP corrections are sufficient at low frequencies (∼1Hz) but temporal white noise is also required at higher frequencies to desensitize penumbral cones. We therefore recommend the selective application of individualized observer calibration and/or temporal white noise in silent-substitution paradigms when studying melanopsin-directed photoresponses.

The accuracy of artificial and natural light measurements by actigraphs.

Actigraphs are the reference standard for measuring light exposure in human non-laboratory experiments due to their portability and long battery lives. However, actigraphs typically have a limited illuminance operating range not representative of real-world conditions, and for many actigraphs, the accuracy of their light measurement has not been verified independently. We assessed the illuminances recorded by Activinsights GENEActiv Original and Philips Actiwatch 2 actigraphs in comparison to a calibrated, laboratory-standard photometer, under both artificial light-emitting diode (LED) and natural sunlight illuminations that might be encountered by a person under real-world conditions. We show that in response to ~20,000 lux white LED light, the GENEActiv and Actiwatch 2 underestimate illuminance by recording 50% and 25% of the true value, respectively. Under ~30,000 lux sunlight, the GENEActiv readily saturates whereas the Actiwatch 2 reports ~46% of the true illuminance. These underestimations are highly linear and we provide correction factors to estimate the illuminance levels of the ambient environment measured by the actigraphs. We also evaluate the application of neutral density filters for extending the operating range of both devices in natural sunlight illuminations (as high as 30,000 lux during our measurements) and demonstrate that this may be a viable approach for increasing the operating range of the Actiwatch 2 but not the GENEActiv. We conclude that both actigraphs provide good performance in monitoring the temporal patterning of light, whereas the absolute illuminance values require correction to accurately evaluate the effects of light intensity on human health and behaviours.

Rhodopsin and melanopsin contributions to human brightness estimation.

We examined the contributions of rhodopsin and melanopsin to human brightness estimation under dim lighting. Absolute brightness magnitudes were estimated for full-field, rhodopsin-, or melanopsin-equated narrowband lights (${\lambda _{\rm max}}:\;{462}$λmax:462, 499, 525 nm). Our data show that in scotopic illumination ($ - {5.1}$-5.1 to $ - {3.9}\;{\log}\;\unicode{x00B5} {\rm Watts}\cdot{\rm cm}^{ - 2}$-3.9logµWatts⋅cm-2), the perceptual brightness estimates of rhodopic irradiance-equated conditions are independent of their corresponding melanopic irradiance, whereas brightness estimates with melanopic irradiance-equated conditions increase with increasing rhodopic irradiance. In mesopic illumination ($ - {3.4}$-3.4 to $ - {1.9}\;{\log}\;\unicode{x00B5} {\rm Watts}\cdot{\rm cm}^{ - 2}$-1.9logµWatts⋅cm-2), the brightness estimates with both lighting conditions increase with increasing rhodopic or melanopic irradiances. Rhodopsin activation therefore entirely signals scotopic brightness perception and plateaus in mesopic illumination where intrinsic melanopsin contributions become first evident. We infer that all photoreceptor signals are transmitted to higher visual centers for representing scene brightness in scotopic and mesopic illumination through both conventional and melanopsin ganglion cell pathways.

Cone and melanopsin contributions to human brightness estimation: reply.

Our analytical description of full-field brightness perception data [J. Opt. Soc. Am. A35, B19 (2018)JOAOD60740-323210.1364/JOSAA.35.000B19] with contributions from cone luminance and melanopsin expressing intrinsically photosensitive retinal ganglion cells has been extended [J. Opt. Soc. Am. A35, 1780 (2018)JOAOD60740-323210.1364/JOSAA.35.001780] to include S-cones through a blue-yellow opponent channel. We welcome this reanalysis and provide a few remarks on the approach.

Extrinsic cone-mediated post-receptoral noise inhibits the rod temporal impulse response function.

We determined how extrinsic white noise correlating with cone inputs to the three primary visual pathways affects both rod-pathway temporal contrast sensitivity and the impulse response function. A four-primary photostimulator provided independent control of rod and cone photoreceptor excitations under mesopic illumination (20 photopic Td). We show that rod-pathway temporal contrast sensitivity uniformly decreases across all temporal frequencies in the presence of cone noise correlating with the inferred magnocellular, parvocellular, or koniocellular pathways. The rod-pathway temporal impulse response functions derived using the Stork-Falk procedure (with a minimum phase assumption) had lower amplitudes in the pathway-specific cone noise. Therefore, cone noise impairs rod-pathway temporal contrast sensitivity without delaying rod-pathway signal transmission.

Correlated cone noise decreases rod signal contributions to the post-receptoral pathways.

This study investigated how invisible extrinsic temporal white noise that correlates with the activity of one of the three [magnocellular (MC), parvocellular (PC), or koniocellular (KC)] post-receptoral pathways alters mesopic rod signaling. A four-primary photostimulator provided independent control of the rod and three cone photoreceptor excitations. The rod contributions to the three post-receptoral pathways were estimated by perceptually matching a 20% contrast rod pulse by independently varying the LMS (MC pathway), +L-M (PC pathway), and S-cone (KC pathway) excitations. We show that extrinsic cone noise caused a predominant decrease in the overall magnitude and ratio of the rod contributions to each pathway. Thus, the relative cone activity in the post-receptoral pathways determines the relative mesopic rod inputs to each pathway.

Cone and melanopsin contributions to human brightness estimation.

We determined the contributions of cone and melanopsin luminance signaling to human brightness perception. The absolute brightness of four narrowband primary lights presented in a full-field Ganzfeld was estimated in two conditions, either cone luminance-equated (186.7-1,867.0 cd·m-2) or melanopsin luminance-equated (31.6-316.3 melanopsin cd·m-2). We show that brightness estimations for each primary light follow an approximately linear increase with increasing cone or melanopsin luminance (in log units), but are not equivalent for primary lights equated with either cone or melanopsin luminance. Instead, brightness estimations result from a combined interaction between cone and melanopsin signaling. Analytical modeling with wavelength-dependent coefficients signifies that melanopsin luminance positively correlates with brightness magnitudes, and the cone luminance has two contribution components, one that is additive to melanopsin luminance and a second that is negative, implying an adaptation process. These results provide a new framework for evaluating the physiological basis of brightness perception and have direct practical applications for the development of energy-efficient light sources.

Correlated and uncorrelated invisible temporal white noise alters mesopic rod signaling.

We determined how rod signaling at mesopic light levels is altered by extrinsic temporal white noise that is correlated or uncorrelated with the activity of one (magnocellular, parvocellular, or koniocellular) postreceptoral pathway. Rod and cone photoreceptor excitations were independently controlled using a four-primary photostimulator. Psychometric (Weibull) functions were measured for incremental rod pulses (50 to 250 ms) in the presence (or absence; control) of perceptually invisible subthreshold extrinsic noise. Uncorrelated (rod) noise facilitates rod detection. Correlated postreceptoral pathway noise produces differential changes in rod detection thresholds and decreases the slope of the psychometric functions. We demonstrate that invisible extrinsic noise changes rod-signaling characteristics within the three retinogeniculate pathways at mesopic illumination depending on the temporal profile of the rod stimulus and the extrinsic noise type.

Melanopsin-mediated post-illumination pupil response in the peripheral retina.

Intrinsically photosensitive retinal ganglion cells (ipRGCs) regulate pupil size by integrating extrinsic rod and cone signals with intrinsic melanopsin-mediated phototransduction. Light adapted pupil diameter is determined by the corneal flux density (CFD), and for central visual field stimulation the melanopsin-mediated post-illumination pupil response (PIPR) follows this same CFD relationship. Rods, cones, and ipRGCs vary in size, density, and distribution across the retina, but how these differences affect the amplitude and timing of the extrinsic and intrinsic pupil light reflex in the central and peripheral retina is unknown. We determined the relationship between stimulus area and photon flux with stimuli constant for CFD, irradiance, or area at central (0°) and peripheral (20°) eccentricities with high and low melanopsin excitation. We show that the pupil constriction amplitude was similar at both eccentricities and the time to minimum diameter increased as melanopsin excitation increased. In contrast, the peripheral PIPR follows a CFD relationship but with lower amplitude compared with that at the fovea. This indicates differences in the spatial and temporal characteristics of extrinsic and intrinsic ipRGC inputs to the pupil control pathway for the central and peripheral retina. The eccentricity-dependent change in PIPR amplitude may be analogous to the hill of vision observed in visual perimetry; such knowledge is an important precursor to the development of pupil perimetry paradigms to measure the PIPR in select regions of the visual field.

Peripheral detection and resolution with mid-/long-wavelength and short-wavelength sensitive cone systems.

This study compared neural resolution and detection limits of the human mid-/long-wavelength and short-wavelength cone systems with anatomical estimates of photoreceptor and retinal ganglion cell spacings and sizes. Detection and resolution limits were measured from central fixation out to 35° eccentricity across the horizontal visual field using a modified Lotmar interferometer. The mid-/long-wavelength cone system was studied using a green (550 nm) test stimulus to which S-cones have low sensitivity. To bias resolution and detection to the short-wavelength cone system, a blue (450 nm) test stimulus was presented against a bright yellow background that desensitized the M- and L-cones. Participants were three trichromatic males with normal visual functions. With green stimuli, resolution showed a steep central-peripheral gradient that was similar between participants, whereas the detection gradient was shallower and patterns were different between participants. Detection and resolution with blue stimuli were poorer than for green stimuli. The detection of blue stimuli was superior to resolution across the horizontal visual field and the patterns were different between participants. The mid-/long-wavelength cone system's resolution is limited by midget ganglion cell spacing and its detection is limited by the size of the M- and L-cone photoreceptors, consistent with previous observations. We found that no such simple relationships occur for the short-wavelength cone system between resolution and the bistratified ganglion cell spacing, nor between detection and the S-cone photoreceptor sizes.

A method for estimating intrinsic noise in electroretinographic (ERG) signals.

PURPOSE: To develop a signal processing paradigm for extracting ERG responses to temporal sinusoidal modulation with contrasts ranging from below perceptual threshold to suprathreshold contrasts and estimate the magnitude of intrinsic noise in ERG signals at different stimulus contrasts. METHODS: Photopic test stimuli were generated using a 4-primary Maxwellian view optical system. The 4-primary lights were sinusoidally temporally modulated in-phase (36 Hz; 2.5-50% Michelson contrast). The stimuli were presented in 1-s epochs separated by a 1-ms blank interval and repeated 160 times (160.160-s duration) during the recording of the continuous flicker ERG from the right eye using DTL fibre electrodes. After artefact rejection, the ERG signal was extracted using Fourier transforms in each of the 1-s epochs where a stimulus was presented. The signal processing allows for computation of the intrinsic noise distribution in addition to the signal-to-noise (SNR) ratio. RESULTS: We provide the initial report that the ERG intrinsic noise distribution is independent of stimulus contrast, whereas SNR decreases linearly with decreasing contrast until the noise limit at ~2.5%. The 1-ms blank intervals between epochs de-correlated the ERG signal at the line frequency (50 Hz) and thus increased the SNR of the averaged response. We confirm that response amplitude increases linearly with stimulus contrast. The phase response shows a shallow positive relationship with stimulus contrast. CONCLUSIONS: This new technique will enable recording of intrinsic noise in ERG signals above and below perceptual visual threshold and is suitable for measurement of continuous rod and cone ERGs across a range of temporal frequencies, and post-receptoral processing in the primary retinogeniculate pathways at low stimulus contrasts. The intrinsic noise distribution may have application as a biomarker for detecting changes in disease progression or treatment efficacy.

Cone ratios in myopia and emmetropia: a pilot study.

PURPOSE: There is a suggestion that the long wavelength-sensitive (LWS)-to-middle wavelength-sensitive (MWS) cone ratio in the retina is associated with myopia. The aim was to measure the LWS/MWS amplitude modulation ratio, an estimate of the LWS/MWS cone ratio, in young adult emmetropes and myopes. METHODS: Multifocal visual evoked potentials were measured when the LWS and MWS cone systems were excited separately using the method of silent substitution. The 30 young adult participants (22 to 33 years) included 10 emmetropes (mean [±SD] refraction, +0.3 [±0.4] diopters [D]) and 20 myopes (mean [±SD] refraction, -3.4 [±1.7] D). RESULTS: The LWS/MWS amplitude modulation ratios ranged from 0.56 to 1.80 in the central 3- to 13-degree diameter ring and from 0.94 to 1.91 in the peripheral 13- to 30-degree diameter ring. Within the central ring, the mean (±SD) ratios were 1.20 (±0.26) and 1.20 (±0.33) for the emmetropic and the myopic groups, respectively. For the peripheral ring, the mean (±SD) ratios were 1.48 (±0.27) and 1.30 (±0.27), respectively. There were no significant differences in the ratios between the emmetropic and myopic groups for either the central (p = 0.99) or peripheral (p = 0.08) rings. For the latter, more myopic refractive error was associated with lower LWS/MWS amplitude modulation ratio; the refraction explained 16% (p = 0.02) of variation in ratio. CONCLUSIONS: The relationship between the LWS/MWS amplitude modulation ratios and refraction at 13 to 30 degrees indicates that a large longitudinal study of changes in refraction in persons with known cone ratio is required to determine if a low LWS/MWS cone ratio is associated with myopia development.

Effect of rod-cone interactions on mesopic visual performance mediated by chromatic and luminance pathways.

We studied the effect of rod-cone interactions on mesopic visual reaction time (RT). Rod and cone photoreceptor excitations were independently controlled using a four-primary photostimulator. It was observed that (1) lateral rod-cone interactions increase the cone-mediated RTs; (2) the rod-cone interactions are strongest when rod sensitivity is maximal in a dark surround, but weaker with increased rod activity in a light surround; and (3) the presence of a dark surround nonselectively increased the mean and variability of chromatic (+L-M, S-cone) and luminance (L+M+S) RTs independent of the level of rod activity. The results demonstrate that lateral rod-cone interactions must be considered when deriving mesopic luminous efficiency using RT.

Melanopsin-expressing intrinsically photosensitive retinal ganglion cells in retinal disease.

Melanopsin-containing intrinsically photosensitive retinal ganglion cells (ipRGCs) are a class of photoreceptors with established roles in non-image-forming processes. Their contributions to image-forming vision may include the estimation of brightness. Animal models have been central for understanding the physiological mechanisms of ipRGC function and there is evidence of conservation of function across species. Intrinsically photosensitive retinal ganglion cells can be divided into five ganglion cell subtypes that show morphological and functional diversity. Research in humans has established that ipRGCs signal environmental irradiance to entrain the central body clock to the solar day for regulating circadian processes and sleep. In addition, ipRGCs mediate the pupil light reflex (PLR), making the PLR a readily accessible behavioral marker of ipRGC activity. Less is known about ipRGC function in retinal and optic nerve disease, with emerging research providing insight into their function in diabetes, retinitis pigmentosa, glaucoma, and hereditary optic neuropathy. We briefly review the anatomical distributions, projections, and basic physiological mechanisms of ipRGCs and their proposed and known functions in animals and humans with and without eye disease. We introduce a paradigm for differentiating inner and outer retinal inputs to the pupillary control pathway in retinal disease and apply this paradigm to patients with age-related macular degeneration (AMD). In these cases of patients with AMD, we provide the initial evidence that ipRGC function is altered and that the dysfunction is more pronounced in advanced disease. Our perspective is that with refined pupillometry paradigms, the PLR can be extended to AMD assessment as a tool for the measurement of inner and outer retinal dysfunction.