Prolonged Inner Retinal Photoreception Depends on the Visual Retinoid Cycle.

In addition to rods and cones, mammals have inner retinal photoreceptors called intrinsically photosensitive retinal ganglion cells (ipRGCs), which use the photopigment melanopsin and mediate nonimage-forming visual responses, such as pupil reflexes and circadian entrainment. After photic activation, photopigments must be reverted to their dark state to be light-sensitive again. For rods and to some extent cones, photopigment regeneration depends on the retinoid cycle in the adjacent retinal pigment epithelium (RPE). By contrast, ipRGCs are far from the RPE, and previous work suggests that melanopsin is capable of light-dependent self-regeneration. Here, we used in vitro ipRGC recording and in vivo pupillometry to show that the RPE is required for normal melanopsin-based responses to prolonged light, especially at high stimulus intensities. Melanopsin-based photoresponses of rat ipRGCs were remarkably sustained when a functional RPE was attached to the retina, but became far more transient if the RPE was removed, or if the retinoid cycle was inhibited, or when Müller glia were poisoned. Similarly, retinoid cycle inhibition markedly reduced the steady-state amplitude of melanopsin-driven pupil reflexes in both mice and rats. However, melanopsin photoresponses in RPE-separated rat retinas became more sustained in the presence of an 11-cis-retinal analog. In conclusion, during prolonged illumination, melanopsin regeneration depends partly on 11-cis-retinal from the RPE, possibly imported via Müller cells. Implications for RPE-related eye diseases and the acne drug isotretinoin (a retinoid cycle inhibitor) are discussed. SIGNIFICANCE STATEMENT: Intrinsically photosensitive retinal ganglion cells (ipRGCs) contain the photopigment melanopsin and drive subconscious physiological responses to light, e.g., pupillary constriction and neuroendocrine regulation. In darkness, each photopigment molecule in ipRGCs, as well as rod/cone photoreceptors, contains 11-cis-retinal (a vitamin A derivative) and light isomerizes it to all-trans-retinal, which activates the photopigment. To make this photopigment excitable again,all-trans-retinal must be reisomerized to 11-cis-retinal. For rods and to some extent cones, this reisomerization occurs in the adjacent retinal pigment epithelium (RPE), but because ipRGCs are far from the RPE, they are thought to regenerate excitable melanopsin exclusively through RPE-independent means. Here, we present electrophysiological and behavioral evidence that ipRGCs depend on the RPE to continuously regenerate melanopsin during intense prolonged photostimulation.

Melatonin Suppression by Light in Humans Is More Sensitive Than Previously Reported.

The retina drives various non-image-forming photoresponses, including circadian photoentrainment and pupil constriction. Previous investigators showed that in humans, photic suppression of the clock-controlled hormone melatonin is most sensitive to 460-nm blue light, with a threshold of ~12 log photons cm(-2) s(-1). This threshold is surprising because non-image-forming vision is mediated by intrinsically photosensitive retinal ganglion cells, which receive rod-driven synaptic input and can respond to light levels as low as ~7 log photons cm(-2) s(-1). Using a protocol that enhances data precision, we have found the threshold for human melatonin suppression to be ~10 log photons cm(-2) s(-1) at 460 nm. This finding has far-reaching implications since there is mounting evidence that nocturnal activation of the circadian system can be harmful.

The rat retina has five types of ganglion-cell photoreceptors.

Intrinsically photosensitive retinal ganglion cells (ipRGCs) are inner retinal photoreceptors that mediate non-image-forming visual functions, e.g. pupillary constriction, regulation of pineal melatonin release, and circadian photoentrainment. Five types of ipRGCs were recently discovered in mouse, but whether they exist in other mammals remained unknown. We report that the rat also has five types of ipRGCs, whose morphologies match those of mouse ipRGCs; this is the first demonstration of all five cell types in a non-mouse species. Through immunostaining and λmax measurements, we showed that melanopsin is likely the photopigment of all rat ipRGCs. The various cell types exhibited diverse spontaneous spike rates, with the M1 type spiking the least and M4 spiking the most, just like we had observed for their mouse counterparts. Also similar to mouse, all ipRGCs in rat generated not only sluggish intrinsic photoresponses but also fast, synaptically driven ones. However, we noticed two significant differences between these species. First, whereas we learned previously that all mouse ipRGCs had equally sustained synaptic light responses, rat M1 cells' synaptic photoresponses were far more transient than those of M2-M5. Since M1 cells provide all input to the circadian clock, this rat-versus-mouse discrepancy could explain the difference in photoentrainment threshold between mouse and other species. Second, rat ipRGCs' melanopsin-based spiking photoresponses could be classified into three varieties, but only two were discerned for mouse ipRGCs. This correlation of spiking photoresponses with cell types will help researchers classify ipRGCs in multielectrode-array (MEA) spike recordings.

Bias-free double judgment accuracy during spatial attention cueing: performance enhancement from voluntary and involuntary attention.

Recent research has demonstrated that involuntary attention improves target identification accuracy for letters using non-predictive peripheral cues, helping to resolve some of the controversy over performance enhancement from involuntary attention. While various cueing studies have demonstrated that their reported cueing effects were not due to response bias to the cue, very few investigations have quantified the extent of any response bias or developed methods of removing bias from observed results in a double judgment accuracy task. We have devised a method to quantify and remove response bias to cued locations in a double judgment accuracy cueing task, revealing the true, unbiased performance enhancement from involuntary and voluntary attention. In a 7-alternative forced choice cueing task using backward masked stimuli to temporally constrain stimulus processing, non-predictive cueing increased target detection and discrimination at cued locations relative to uncued locations even after cue location bias had been corrected.

Bias corrected double judgment accuracy during spatial attention cueing: unmasked stimuli with non-predictive and semi-predictive cues.

The present experiments indicate that in a 7-AFC double judgment accuracy task with unmasked stimuli, cue location response bias can be quantified and removed, revealing unbiased improvements in response accuracy for valid cues compared to invalid cues. By testing for cueing effects over a range of contrast levels with unmasked stimuli, changes in the psychometric function were examined and provide insight into the mechanisms of involuntary attention which might account for the observed cueing effects. Cue validity was varied between two separate experiments showing that non-predictive (14.3%) and moderately-predictive cues (50%) equally facilitate stimulus identification and localization during transient involuntary attention capture. Observers had improved accuracy at identifying both the location and the feature identity of target letters throughout a range of contrast levels, without any dependence on backward masking. There was a leftward shift of the psychometric function threshold with valid cued data and no slope reduction suggesting that any additive hypothesis based on spatial uncertainty reduction or perceptual enhancement is not a sufficient explanation for the observed cueing effects. The interdependence of the perceptual processes of stimulus discrimination and localization were also investigated by analyzing response contingencies, showing that observers were equally skilled at making identification and localization accuracy judgments with unmasked stimuli.

Involuntary attention enhances identification accuracy for unmasked low contrast letters using non-predictive peripheral cues.

There is controversy regarding whether or not involuntary attention improves response accuracy at a cued location when the cue is non-predictive and if these cueing effects are dependent on backward masking. Various perceptual and decisional mechanisms of performance enhancement have been proposed, such as signal enhancement, noise reduction, spatial uncertainty reduction, and decisional processes. Herein we review a recent report of mask-dependent accuracy improvements with low contrast stimuli and demonstrate that the experiments contained stimulus artifacts whereby the cue impaired perception of low contrast stimuli, leading to an absence of improved response accuracy with unmasked stimuli. Our experiments corrected these artifacts by implementing an isoluminant cue and increasing its distance relative to the targets. The results demonstrate that cueing effects are robust for unmasked stimuli presented in the periphery, resolving some of the controversy concerning cueing enhancement effects from involuntary attention and mask dependency. Unmasked low contrast and/or short duration stimuli as implemented in these experiments may have a short enough iconic decay that the visual system functions similarly as if a mask were present leading to improved accuracy with a valid cue.