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Optineurin (OPTN) is a gene associated with familial normal tension glaucoma (NTG). While NTG involves intraocular pressure (IOP)-independent neurodegeneration of the visual pathway that progresses with age, how OPTN dysfunction leads to NTG remains unclear. Here, we generated an OPTN knockout mouse (Optn -/ - ) model to test the hypothesis that a loss-of-function mechanism induces structural and functional eye deterioration with aging. Eye anatomy, visual function, IOP, retinal histology, and retinal ganglion cell survival were compared to littermate wild-type (WT) control mice. Consistent with OPTN's role in NTG, loss of OPTN did not increase IOP or alter gross eye anatomy in young (2-3 months) or aged (12 months) mice. When retinal layers were quantitated, young Optn -/ - mice had thinner retina in the peripheral regions than young WT mice, primarily due to thinner ganglion cell-inner plexiform layers. Despite this, visual function in Optn -/ - mice was not severely impaired, even with aging. We also assessed relative abundance of retinal cell subtypes, including amacrine cells, bipolar cells, cone photoreceptors, microglia, and astrocytes. While many of these cellular subtypes were unaffected by Optn deletion, more dopaminergic amacrine cells were observed in aged Optn -/ - mice. Taken together, our findings showed that complete loss of Optn resulted in mild retinal changes and less visual function impairment, supporting the possibility that OPTN-associated glaucoma does not result from a loss-of-function disease mechanism. Further research using these Optn mice will elucidate detailed molecular pathways involved in NTG and identify clinical or environmental risk factors that can be targeted for glaucoma treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Effective strategies for the neuroprotection and preservation of retinal ganglion cells (RGCs) remain elusive in the management of glaucoma. A spontaneous genetic model of glaucoma has been identified in cats and extensively characterized as a viable translational model, with eye size and anatomy similar to humans. In this study we sought to establish initial proof of concept for gene delivery to feline RGCs via intravitreal injection of AAV2 in normal cats. Pre-retinal, posterior vitreal injection of AAV2/2-CMV-GFP, was performed overlying the area centralis in 5 adult cats. Immunosuppressive oral prednisolone was administered perioperatively and gradually tapered over 6-10wks post-injection. Ophthalmic examination was performed pre- and post-injection. The GFP reporter expression and morphological effects of viral transduction on the retina were monitored in vivo using confocal scanning laser ophthalmoscopy (cSLO) and optical coherence tomography (OCT), respectively (Spectralis OCT-HRA, Heidelberg), at 1-2wk intervals over 6-10wks. Full-field electroretinograms (ERG) and visual evoked potentials (VEP) were recorded at baseline and post-injection. Retinas were examined by histology and immunolabeling for the RGC marker RBPMS and Müller cell and astrocyte marker SOX9, and GFP expression was examined in the retina, optic nerve (ON), optic tract and lateral geniculate nucleus (LGN). GFP + retinal cells and RGC axons were visualized by cSLO at 1-2 weeks post-injection. No retinal morphological changes were observed by OCT in vivo but 3/5 eyes exhibited mild retinal inflammation on histology. Retinal and ON function were preserved in injected eyes compared to baseline and untreated eyes. GFP expression was predominantly identified in RBPMS + RGC cells as well as SOX9 + Müller cells. GFP fluorescence was observed throughout RGC nerve fiber tract in the central visual pathway. Peak transduction in RGCs (up to ∼ 20 %) was observed in the regions with high GFP expression, but < 1 % of RGCs expressed GFP across the whole retina. Our data provide proof of concept that pre-retinal injection of AAV2/2 may represent a feasible platform for gene delivery to feline RGCs in vivo but highlight a need for further refinement to improve RGC transduction efficiency and control low-grade retinal inflammation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The temporal resolution of the visual system progressively increases with light intensity. Under scotopic conditions, temporal resolution is relatively poor, and may be limited by both retinal and cortical processes. Rod photoresponses themselves are quite slow because of the slowly deactivating biochemical cascade needed for light transduction. Here, we have used a transgenic mouse line with faster than normal rod phototransduction deactivation (RGS9-overexpressors) to test whether rod signaling to second-order retinal neurons is rate-limited by phototransduction or by other mechanisms. We compared electrical responses of individual wild-type and RGS9-overexpressing (RGS9-ox) rods to steady illumination and found that RGS9-ox rods required 2-fold brighter light for comparable activation, owing to faster G-protein deactivation. When presented with flickering stimuli, RGS9-ox rods showed greater magnitude fluctuations around a given steady-state current amplitude. Likewise, in vivo electroretinography (ERG) and whole-cell recording from OFF-bipolar, rod bipolar, and horizontal cells of RGS9-ox mice displayed larger than normal magnitude flicker responses, demonstrating an improved ability to transmit frequency information across the rod synapse. Slow phototransduction recovery therefore limits synaptic transmission of increments and decrements of light intensity across the first retinal synapse in normal retinas, apparently sacrificing temporal responsiveness for greater overall sensitivity in ambient light.

The skate retina provides a native all-rod retina suited for investigating a single type of photoreceptor regarding its properties and signaling to second order cells. Using the aspartate-induced isolated A-wave of the skate eyecup electroretinogram (ERG), it has been shown that adaptation in rods remains Weber-Fechner-like over a 6-log unit increase in background light intensity. Zinc, which can block calcium channels, has been found in the rod synaptic terminal and the synaptic cleft. Histidine is a zinc chelator. Voltage signals from neurons post-synaptic to rods indicate that histidine increases the dark release of glutamate and increases the horizontal cell light response. In histidine, the A-wave response to various light intensities in the dark-adapted retina increased more than fifty percent, corresponding to the effect on horizontal cells. In the presence of background light, although histidine-treated rod light responses remained Weber-Fechner-like, their increment threshold was raised significantly. This indicates that endogenous zinc feedback serves to increase rod sensitivity in a light-adapted retina, despite a corresponding reduction of threshold sensitivity in the dark. We propose that the increase in A-wave amplitude is a result of the increased conductance at the synaptic terminal and that the A-wave can be used to monitor changes in rod transmitter release. Furthermore, endogenous zinc may also provide the benefit of reducing metabolic stress and the risk of glutamate toxicity in the dark. [ABSTRACT FROM AUTHOR]

Recent reports indicate that the subjective ability of humans to discriminate between polarization E-vector orientations approaches that of many invertebrates. Here, we show that polarization-modulated patterned stimuli generate an objectively recordable electrophysiological response in humans with normal vision. We investigated visual evoked potential (VEP) and electroretinographic (ERG) responses to checkerboard patterns defined solely by their polarization E-vector orientation alternating between ± 45°. Correcting for multiple comparisons, paired-samples t-tests were conducted to assess the significance of post-stimulus deflections from baseline measures of noise. Using standard check pattern sizes for clinical electrophysiology, and a pattern-reversal protocol, participants showed a VEP response to polarization-modulated patterns (PolVEP) with a prominent and consistent positive component near 150 ms (p < 0.01), followed by more variable negative components near 200 ms and 300 ms. The effect was unrecordable with visible wavelengths >550 nm. Further, pseudo-depolarization negated the responses, while control studies provided confirmatory evidence that the PolVEP response was not the product of luminance artefacts. Polarization-modulated patterns did not elicit a recordable ERG response. The possible origins of the PolVEP signals, and the absence of recordable ERG signals, are discussed. We conclude that evoked cortical responses to polarization-modulated patterns provide an objective measure of foveal function, suitable for both humans and non-human primates with equivalent macular anatomy. [ABSTRACT FROM AUTHOR]

To study the physiology of the primate visual system, non-invasive electrophysiological techniques are of major importance. Two main techniques are available: the electroretinogram (ERG), a mass potential originating in the retina, and the visual evoked potential (VEP), which reflects activity in the primary visual cortex. In this overview, the history and the state of the art of these techniques are briefly presented as an introduction to the special issue "New Developments in non-invasive visual electrophysiology". The overview and the special issue can be used as the starting point for exciting new developments in the electrophysiology of primate and mammalian vision. [ABSTRACT FROM AUTHOR]

Most animals rely on vision to perform a range of behavioural tasks and variations in the anatomy and physiology of the eye likely reflect differences in habitat and life history. Moreover, eye design represents a balance between often conflicting requirements for gathering different forms of visual information. The trade-off between spatial resolving power and contrast sensitivity is common to all visual systems, and European honeybees (Apis mellifera) present an important opportunity to better understand this trade-off. Vision has been studied extensively in A. mellifera as it is vital for foraging, navigation and communication. Consequently, spatial resolving power and contrast sensitivity in A. mellifera have been measured using several methodologies; however, there is considerable variation in estimates between methodologies. We assess pattern electroretinography (pERG) as a new method for assessing the trade-off between visual spatial and contrast information in A.mellifera. pERG has the benefit of measuring spatial contrast sensitivity from higher order visual processing neurons in the eye. Spatial resolving power of A.mellifera estimated from pERG was 0.54 cycles per degree (cpd), and contrast sensitivity was 16.9. pERG estimates of contrast sensitivity were comparable to previous behavioural studies. Estimates of spatial resolving power reflected anatomical estimates in the frontal region of the eye, which corresponds to the region stimulated by pERG. Apis mellifera has similar spatial contrast sensitivity to other hymenopteran insects with similar facet diameter (Myrmecia ant species). Our results support the idea that eye anatomy has a substantial effect on spatial contrast sensitivity in compound eyes. [ABSTRACT FROM AUTHOR]

The idea of implanting microphotodiode arrays as visual prostheses has aroused controversy on its feasibility from the moment it appeared in print. We now present results which basically support the concept of replacing damaged photoreceptors with subretinally implanted stimulation devices. Network activity in degenerated rat retinae could be modulated through local electrical stimulation in vitro. We also investigated the long term stability and biocompatibility of the subretinal implants and their impact on retinal physiology in rats. Ganzfeld electroretinograms and histology showed no significant side effect of subretinal implants on retinal function or the architecture of the inner retina.

The white noise electroretinogram (wnERG) provides a measure of the impulse response function under conditions of retinal equilibrium; it is yet to be determined how the electrical response generated by melanopsin ganglion cell photoreception is expressed in the impulse response. To this end, we recorded the human wnERG to continuous temporal white noise (TWN) stimuli that were melanopsin-directed (rod and cone silent) or cone-directed (rod and melanopsin silent). The impulse response of the electroretinogram was derived by cross-correlating the TWN stimulus with the wnERG response. We observed that the LMS-cone directed wnERG contained the expected N1 wave (24.1 ± 2.4 ms; mean ± SEM) and P1 wave (49.7 ± 1.8 ms). Melanopsin-directed stimuli produced a unique wnERG with a slower negative deflection (Nm) at 62.9 ± 3.3 ms followed by a positive deflection (Pm) at 126.3 ± 5.1 ms. Additional experiments indicated this melanopsin-directed wnERG response was not due to cone intrusion. The Nm and NmPm amplitudes increased with illuminance (32,000-80,000 Td; no rod intrusion) and melanopsin contrast (10-36% Michelson contrast). As there are known pathways connecting melanopsin cells to the outer retina, we then measured the wnERG to combined melanopsin and cone-directed stimuli to quantify melanopsin interactions with cone signalling. With the combined stimuli, the N1P1 amplitudes were suppressed by ~59%, which may be a result of a destructive interference between the positive (P1) and negative (Nm) waves generated by the cone and melanopsin pathways. We conclude that the human wnERG to melanopsin-directed stimuli may reflect the combined response of intra-retinal melanopsin pathways, independent of rod and cone photoreception. [ABSTRACT FROM AUTHOR]

The flicker electroretinogram (ERG) is typically analyzed in terms of peak-to-trough amplitude and implicit time. However, additional important information may be captured by spectral-domain analysis of the ERG harmonics (responses that occur at multiples of the stimulus frequency). This study describes an approach to analyze the harmonic components of the flicker ERG and its application to patients who have early-stage non-proliferative diabetic retinopathy (NPDR). Of particular interest were the sub-harmonic components occurring at 1.5x and 2.5x the stimulus frequency that produce cycle-to-cycle variation in amplitude termed "period doubling." Twenty visually-normal subjects, 20 diabetic subjects who have no clinically-apparent retinopathy (NDR), and 20 diabetic subjects who have mild NPDR participated. ERGs were recorded in response to sinusoidal flicker (27-63 Hz) and Fourier analysis was performed to extract fundamental and harmonic response amplitudes. Linear quantile mixed models (LQMMs) were used to compare the amplitude of the response components among the three subject groups. Results indicated that the maximum sub-harmonic amplitude occurred in the stimulus frequency range of 33-38 Hz for all subjects. The LQMMs showed a significant sub-harmonic amplitude reduction for the mild NPDR subjects compared to the controls; sub-harmonic amplitude for the NDR subjects did not differ significantly from the controls. In contrast, the fundamental response did not differ among the groups for stimulus frequencies between 33 and 38 Hz. Modeling these results indicated that subharmonic amplitude loss in mild NPDR subjects may be attributed to attenuated feedback occurring early in the retina, possibly at the synapse of cone photoreceptors and OFF bipolar cells. [ABSTRACT FROM AUTHOR]

Electroretinograms (ERGs) elicited by high temporal frequency (26-95 Hz) L- and M-cone isolating sine-wave stimuli were investigated in human observers for full-field (FF) and different spatially restricted stimulus sizes (70°, 50°, 30°, and 10° diameter). Responses to L- and M-cone isolating FF stimuli were maximal around 48 Hz and decreased gradually with increasing temporal frequency up to 95 Hz. The response maximum was shifted to about 30-32 Hz for both L- and M-cone driven responses obtained with spatially restricted stimuli. The M-cone driven responses could only be measured up to 54 Hz with 70° stimuli. The response amplitudes for L- and M-cones and L-/M-cone amplitude ratios decreased with decreasing stimulus size. The ERG response phases to L- and M-cone isolating stimuli decreased with increasing temporal frequency and were about -160° apart for all stimulus sizes up to 34 Hz. Further increase in the temporal frequency displayed a positive correlation between stimulus size and L-M phase difference. The ERG data indicate that the responses evoked by high temporal frequency cone isolating stimuli reflect two mechanisms, one that is more centrally located and displays a maximum at about 30-32 Hz and a peripheral mechanism that is sensitive to higher temporal modulations. We propose that the peripheral mechanism (FF ERGs) reflects magnocellular activity, whereas the central mechanism (ERGs with spatially restricted stimuli) is based on a parvocellular activity up to about 30 Hz. [ABSTRACT FROM AUTHOR]

Retinal and cortical signals initiated by a single cone type can be recorded using the spectral compensation (or silent substitution) paradigm. Moreover, responses to instantaneous excitation increments combined with gradual excitation decreases are dominated by the response to the excitation increment. Similarly, the response to a sudden excitation decrement dominates the overall response when combined with a gradual excitation increase. Here ERGs and VEPs were recorded from 34 volunteers [25.9 ± 10.4 years old (mean ± 1 SD); 25 males, 9 females] to sawtooth flicker (4 Hz) stimuli that elicited L- or M-cone responses using triple silent substitution. The mean luminance (284 cd/m2) and the mean chromaticity (x = 0.5686, y = 0.3716; CIE 1931 color space) remained constant and thus the state of adaptation was the same in all conditions. Color discrimination thresholds along protan, deutan, and tritan axes were obtained from all participants. Dichromatic subjects were genetically characterized by molecular analysis of their opsin genes. ERG responses to L-cone stimuli were absent in protanopes whereas ERG responses to M-cone stimuli were strongly reduced in deuteranopes. Dichromats showed generally reduced VEP amplitudes. Responses to cone-specific stimuli obtained with standard electrophysiological methods may give the same classification as that obtained with the Cambridge Colour Test and in some cases with the genetic analysis of the L- and M-opsin genes. Therefore, cone-specific ERGs and VEPs may be reliable methods to detect cone dysfunction. The present data confirm and emphasize the potential use of cone-specific stimulation, combined with standard visual electrodiagnostic protocols. [ABSTRACT FROM AUTHOR]

Optical imaging of retinal intrinsic signals is a relatively new method that provides spatiotemporal patterns of retinal activity through activity-dependent changes in light reflectance of the retina. The exact physiological mechanisms at the origin of retinal intrinsic signals are poorly understood and there are significant inter-species differences in their characteristics and cellular origins. In this study, we re-examined this issue through pharmacological dissection of retinal intrinsic signals in the rabbit with simultaneous ERG recordings. Retinal intrinsic signals faithfully reflected retinal activity as their amplitude was strongly associated with stimulation intensity (r2=0.85). Further, a strong linear relation was found using linear regression (r2=0.98) between retinal intrinsic signal amplitude and the ERG b wave, which suggests common cellular origins. Intravitreal injections of pharmacological agents were performed to isolate the activity of the retina's major cell types. Retinal intrinsic signals were abolished when the photoreceptors' activity was isolated with aspartate, indicative that they are not at the origin of this signal. A small but significant decrease in intrinsic response (20%) was observed when ganglion and amacrine cells' activity was inhibited by TTX injections. The remaining intrinsic responses were abolished in a dose-dependent manner through the inhibition of ON-bipolar cells by APB. Our results indicate that, in rabbits, retinal intrinsic signals reflect stimulation intensity and originate from the inner retina with a major contribution of bipolar cells and a minor one from ganglion or amacrine cells. [ABSTRACT FROM AUTHOR]

[Display omitted] Autosomal dominant retinitis pigmentosa (adRP) is frequently caused by mutations in RHO , the gene for rhodopsin. In previous experiments in dogs with the T4R mutation in RHO , an AAV2/5 vector expressing an shRNA directed to human and dog RHO mRNA and an shRNA-resistant human RHO cDNA (AAV- RHO820-shRNA820) prevented retinal degeneration for more than eight months following injection. It is crucial, however, to determine if this RNA replacement vector acts in a mutation-independent and species-independent manner. We, therefore, injected mice transgenic for human P23H RHO with this vector unilaterally at postnatal day 30. We monitored their retinal structure by using spectral-domain optical coherence tomography (SD-OCT) and retinal function using electroretinography (ERG) for nine months. We compared these to P23H RHO transgenic mice injected unilaterally with a control vector. Though retinas continued to thin over time, compared to control injected eyes, treatment with AAV-RHO820-shRNA820 slowed the loss of photoreceptor cells and the decrease in ERG amplitudes during the nine-month study period. Unexpectedly, we also observed the preservation of retinal structure and function in the untreated contralateral eyes of AAV- RHO820-shRNA820 treated mice. PCR analysis and western blots showed that a low amount of vector from injected eyes was present in uninjected eyes. In addition, protective neurotrophic factors bFGF and GDNF were elevated in both eyes of treated mice. Our finding suggests that using this or similar RNA replacement vectors in human gene therapy may provide clinical benefit to both eyes of patients with adRP. [ABSTRACT FROM AUTHOR]

Congenital stationary night blindness (CSNB) is a group of inherited retinal diseases in which either rod-to-ON-bipolar cell (ON-BC) signaling, or rod function is affected leading to impaired vision under low light conditions. One type of CSNB is associated with defects in genes (NYX, GRM6, TRPM1, GPR179, and LRIT3) involved in the mGluR6 signaling cascade at the ON-BC dendritic tips. We have previously characterized a canine model of LRIT3-CSNB and demonstrated short-term safety and efficacy of an ON-BC targeting AAV-LRIT3 (AAV K9#4 -shGRM6-cLRIT3-WPRE) gene therapy. Herein, we demonstrate long-term functional recovery and molecular restoration following subretinal injection of the ON-BC targeting AAV-LRIT3 vector in all eight treated eyes for up to 32 months. Following subretinal administration of the therapeutic vector, expression of the LRIT3 transgene, as well as restoration of mGluR6 signaling cascade member TRPM1, were confirmed in the outer plexiform layer (OPL) of the treated area. However, further investigation of the transgene LRIT3 transcript expression by RNA in situ hybridization (RNA-ISH) revealed off-target expression in non-BCs including the photoreceptors, inner nuclear, and ganglion cell layers, despite the use of a mutant AAV K9#4 capsid and an improved mGluR6 promoter designed to specifically transduce and promote expression in ON-BCs. While the long-term therapeutic potential of AAV K9#4 -shGRM6-cLRIT3-WPRE is promising, we highlight the necessity for further optimization of AAV-LRIT3 therapy in the canine CSNB model prior to its clinical application., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The chick is widely used in studies of eye growth regulation and myopia. The aim of this study was to explore the utility of pattern (p)ERG as a tool to assess retinal function in such studies. Effects of optical defocus and diffusing blur, manipulations used to alter eye growth experimentally, were evaluated. PERGs were recorded from White-Leghorn chickens, using a checkerboard pattern, including 8 spatial frequencies (0.05-2.2c/d SF), 13 contrast levels (1-100%), and 8 temporal reversal frequencies (0.5-20Hz). The acute effects of defocus and diffusing blur were examined. Flash- and pERGs were also recorded from chicks that underwent monocular optic nerve section (ONS), to explore the contribution of retinal ganglion cells (RGCs). Measurements were made up to 6weeks post-ONS, complemented with SD-OCT imaging. In normal chicks, the response to 1Hz, 100% contrast stimuli showed positive- and negative-going waveforms at 43ms (P1) and 75ms (N95), respectively, with 0.06-0.1c/d SF eliciting the largest P1 amplitudes of 21.9±2.5μV. Contrast levels above 5% yielded measurable P1 responses. Responses were transient and monophasic for 0.5-5Hzreversal rates, with higher temporal frequencies yielding steady state responses. Defocus and diffusing blur decreased pERG amplitude across all SFs. pERG responses remained normal after ONS, despite the loss of RGCs. In conclusion, chicks show robust pERG responses, which are attenuated by defocus and diffusing blur. The pERG response is not affected by ONS, suggesting that RGCs do not contribute to the chick pERG. [ABSTRACT FROM AUTHOR]

A remarkable feature of neuronal glutamate transporters (EAATs) is their dual functions of classical carriers and ligand-gated chloride (Cl − ) channels. Cl − conductance is rapidly activated by glutamate in subtype EAAT5, which mediates light responses in depolarizing bipolar cells (DBC) in retinae of lower vertebrates. In this study, we examine whether EAAT5 also mediates the DBC light response in mouse. We took advantage of an infrared illuminated micro-injection system, and studied the effects of the EAAT blocker (TBOA) and a glutamate receptor agonist (LAP4) on the mouse electroretinogram (ERG) b-wave responses. Our results showed that TBOA and LAP4 shared similar temporal patterns of inhibition: both inhibited the ERG b-wave shortly after injection and recovered with similar time courses. TBOA inhibited the b-wave completely at mesopic light intensity with an IC50 value about 1 log unit higher than that of LAP4. The inhibitory effects of TBOA and LAP4 were found to be additive in the photopic range. Furthermore, TBOA alone inhibited the b-wave in the cone operative range in knockout mice lacking DBC R s at a low concentration that did not alter synaptic glutamate clearance activity. It also produced a stronger inhibition than that of LAP4 on the cone-driven b-wave measured with a double flash method in wildtype mice. These electrophysiological data suggest a significant role for EAAT5 in mediating cone-driven DBC light responses. Our immunohistochemistry data indicated the presence of postsynaptic EAAT5 on some DBC C s and some DBC R s, providing an anatomical basis for EAAT5’s role in DBC light responses. [ABSTRACT FROM AUTHOR]

n order to evaluate the site of action of picrotoxin (antagonist of ionotropic GABA receptors) on the electroretinographic (ERG) b- and d-waves, in this study we compared its effects on the intensity-response function of the ERG waves in intact light adapted frog eyecup preparations with its effects in eyecups, where the activity of proximal neurons was blocked by 1 mM N-methyl-d-aspartate (MNDA). Picrotoxin markedly enhanced the b- and d-wave amplitude and slowed the time course of the responses at all stimulus intensities in the intact eyecups. Perfusion with NMDA alone caused significant enhancement of the b-wave amplitude and diminution of the d-wave amplitude without altering their time course in the entire intensity range. When picrotoxin was applied in combination with NMDA, an enhancement of the b-wave amplitude and slowing of its time course were observed at all stimulus intensities. The increase of the b-wave amplitude was significantly higher than that seen in NMDA group. Combined application of picrotoxin and NMDA caused an enhancement of the d-wave amplitude at the lower stimulus intensities and its diminution at the higher ones, while the d-wave time course was delayed over the entire intensity range. The results obtained indicate that a part of picrotoxin effects on the amplitude and time course of the photopic ERG b- and d-waves are due to its action in the distal frog retina. [ABSTRACT FROM AUTHOR]

The Function of the retina and effects of drugs on it can be assessed by recording transretinal voltage across isolated retina that is perfused with physiological medium. However, building ex vivo ERG apparatus requires substantial amount of time, resources and expertise. Here we adapted a commercial in vivo ERG system for transretinal ERG recordings from rod and cone photoreceptors and compared rod and cone signaling between ex vivo and in vivo environments. We found that the rod and cone a- and b-waves recorded with the transretinal ERG adapter and a standard in vivo ERG system are comparable to those obtained from live anesthetized animals. However, ex vivo responses are somewhat slower and their oscillatory potentials are suppressed as compared to those recorded in vivo. We found that rod amplification constant (A) was comparable between ex vivo and in vivo conditions, ~10-30 s-2 depending on the choice of response normalization. We estimate that the A in cones is between 3 and 6 s-2 in ex vivo conditions and by assuming equal A in vivo we arrive to light funnelling factor of 3 for cones in the mouse retina. The ex vivo ERG adapter provides a simple and affordable alternative to designing a custom-built transretinal recordings setup for the study of photoreceptors. Our results provide a roadmap to the rigorous quantitative analysis of rod and cone responses made possible with such a system. [ABSTRACT FROM AUTHOR]

Highlights: [•] We examined the impact of high altitude on blue cone function. [•] Blue cone function was examined objectively using electroretinography. [•] At 4559m an attenuation of blue cone function was observed. [•] Functional impairment was not associated with symptoms of acute mountain sickness. [•] First report of blue cone ERG at high altitude. [ABSTRACT FROM AUTHOR]

Abstract: The purpose of this study was to investigate the effect of optical defocus on changes of electrical response as a function of retinal region. Twenty-three subjects (aged 19–25year) with normal ocular health were recruited for global flash multifocal electroretinogram (mfERG) recordings under control (fully corrected) condition, and short-term positive defocus (+2D and +4D) and negative defocus (−2D and −4D) conditions. The amplitudes and implicit times of direct (DC) and induced (IC) components of mfERG responses were pooled into six concentric rings for analyses. The mfERG responses demonstrated more significant changes in amplitude in paracentral retinal regions than in the central regions under defocused conditions. The paracentral DC amplitude showed a significant reduction under negative defocus conditions. In contrast, the paracentral IC amplitude showed a significant increment under positive defocus conditions. Interestingly, the central IC response showed significant reduction in amplitude only to negative defocus, while increasing its amplitude to positive defocus. However, the DC and IC implicit times were virtually unaffected under defocused conditions. Our findings suggest that human retina is able to differentiate defocused signals and to identify positive and negative defocus. It shows that paracentral retina reacts more vigorously to optical defocus than does central retina. [Copyright &y& Elsevier]

Abstract: The purpose of the current study was to determine the progress of cortical functional degeneration in the Royal College of Surgeons (RCS) rat. Cortical responses were measured with optical imaging of intrinsic signals using gratings of various spatial frequencies. Subsequently, electrophysiological recordings were also taken across cortical layers in response to a pulse of broad-spectrum light. We found significant degeneration in the cortical processing of visual information as early as 4weeks of age. These results show that degeneration in the cortical response of the RCS rat starts before development has been properly completed. [Copyright &y& Elsevier]

Abstract: The goal of this pilot study was to assess the effects of acute hypoglycemia on retinal function and contrast sensitivity in individuals with and without diabetes. Hyperinsulinemic hypoglycemic and euglycemic clamp procedures were performed in subjects without diabetes (n =7) and with controlled type 1 diabetes (n =5). Mean age was 28years, and none had retinal disease. During euglycemia (glucose 95–110mg/dl) and acute hypoglycemia (glucose 50–55mg/dl), contrast sensitivity was measured and spatial retinal responses were recorded with multifocal electroretinograms (mfERG), a rapid technique for mapping sensitivity from the foveal, macular and peripheral areas of the retina. During hypoglycemia, retinal responses (mfERG P1 wave) were decreased in both type 1 diabetes subjects and subjects without diabetes. The dominant effect was in the amplitude of the responses in the central macular retina, not in their temporal properties. Responses from the central region, central 100, were on average 1.8-fold lower than those from the periphery for both groups. All diabetes subjects and 3/7 without diabetes reported central scotoma. Decreases in mfERG amplitude were accompanied by decreases in contrast sensitivity. These changes were immediately reversed with the restoration of euglycemia. Overall, this study demonstrates that the acute effects of hypoglycemia in the human eye predominantly involve central vision, and these visual effects originate, at least in part, in the retina. The association between low blood glucose levels and impaired central vision underscores the importance of avoiding when possible and promptly treating hypoglycemia, particularly in individuals with diabetes. [Copyright &y& Elsevier]

Abstract: The effect of dopamine D1 receptor blockade by SCH 23390 on the V–log I function of the ERG b- and d-waves was investigated in dark and light adapted frog eyes. We obtained that the blocker enhanced the amplitude of the b- and d-waves in both conditions of adaptation. The enhancing effect of the blocker was more pronounced on the rod- than cone-dominated responses for the both ERG waves. The absolute sensitivity of the b-wave was not altered, but that of the d-wave was significantly increased. The intensity-response function of the b-wave, but not that of the d-wave, was shifted to the left along the intensity axis. The b-wave V–log I function had steeper slope and narrower dynamic range in both dark and light adapted eyes after the D1 receptor blockade. The results obtained indicate that the endogenous dopamine, acting through D1 receptors, does not play a crucial role in the process of retinal adaptation, although it changes in a specific manner the intensity-response function of both the ERG b- and d-waves. [Copyright &y& Elsevier]

Abstract: Previous studies have proposed that the inner retina is affected in myopes. This study aimed to investigate the changes in adaptive circuitry of the inner retina in myopia, using the global flash multifocal electroretinogram (global flash mfERG) with different levels of contrast (luminance modulation). Fifty-four myopes had global flash mfERG recorded with different contrasts. The direct component (DC) and the induced component (IC) of the mfERG response were pooled into six regions for analysis. The response amplitudes and implicit times at different contrasts were also analysed. Results showed that myopes had significant reduction in the paracentral DC amplitude for the 29% and 49% contrasts and in the paracentral IC amplitude at all contrasts measured. The peripheral IC amplitude for the 49% contrast was also reduced. No significant change was found in implicit time for either DC or IC response. Refractive error explained about 14% of the variance in DC and 16% of the variance in IC amplitude respectively; axial length could not account for additional variance in either paracentral DC or IC amplitudes in the hierarchical regression models used. We concluded that the paracentral retinal region in myopes showed signs of impaired retinal adaptation, suggesting a functional loss at the inner retinal layer. In addition, functions attributed to the outer retinal layer showed only small changes due to myopia. [ABSTRACT FROM AUTHOR]

Abstract: Purpose: To evaluate the impact of intraocular (IOP) reduction on retinal ganglion cell (RGC) function measured using pattern electroretinogram optimized for glaucoma (PERGLA) in glaucoma suspect and glaucomatous eyes receiving latanoprost 0.005% versus placebo. Methods: This was a prospective, placebo-controlled, double masked, cross-over clinical trial. One randomly selected eye of each subject meeting eligibility criteria was enrolled. At each visit, subjects underwent five diurnal measurements between 8:00am and 4:00pm consisting of Goldmann IOP, and PERGLA measurements. A baseline examination was performed following a 4-week washout period, and repeat examination after randomly receiving latanoprost or placebo for 4-weeks. Subjects were then crossed over to receive the alternative therapy for 4weeks following a second washout period, and underwent repeat examination. Linear mixed-effect models were used for the analysis. Results: Sixty-eight eyes (35 glaucoma, 33 glaucoma suspect) of 68 patients (mean age 67.4±10.6years) were enrolled. The mean IOP (mmHg) after latanoprost 0.005% therapy (14.9±3.8) was significantly lower than baseline (18.8±4.7, p <0.001) or placebo (18.0±4.3), with a mean reduction of −20±13%. Mean PERGLA amplitude (μV) and phase (π-radian) using latanoprost (0.49±0.22 and 1.71±0.22, respectively) were similar (p >0.05) to baseline (0.49±0.24 and 1.69±0.19) and placebo (0.50±0.24 and 1.72±0.23). No significant (p >0.05) diurnal variation in PERGLA amplitude was observed at baseline, or using latanoprost or placebo. Treatment with latanoprost, time of day, and IOP were not significantly (p >0.05) associated with PERGLA amplitude or phase. Conclusion: Twenty percent IOP reduction using latanoprost monotherapy is not associated with improvement in RGC function measured with PERGLA. [ABSTRACT FROM AUTHOR]

Abstract: The electroretinogram (ERG) is a non-invasive indicator of retinal function. Light flashes evoke a cornea-negative a-wave followed by a cornea-positive b-wave. Light-adaptation is known to increase the amplitude of cone-dependent b-waves. To identify the underlying mechanism, we recorded rat cone photoresponses in situ, using intravitreally-injected glutamate to block synaptic transmission and intense paired-flash stimuli to isolate cone a-waves. Steady adapting illumination caused a progressive increase in cone a-wave amplitude, which was suppressed in a dose-dependent manner by intravitreal CPTIO, a nitric oxide scavenger. We conclude that light-adaptation causes release of nitric oxide, which enhances the cone photoresponse. [ABSTRACT FROM AUTHOR]

Abstract: Optic nerve transection (ONT) triggers retinal ganglion cell (RGC) death. By using this paradigm, we have analyzed for the first time in adult albino and pigmented mice, the effects of ONT in the scotopic threshold response (STR) components (negative and positive) of the full-field electroretinogram. Two weeks after ONT, when in pigmented mice approximately 18% of the RGC population survive, the STR-implicit time decreased and the p and nSTR waves diminished approximately to 40% or 55%, in albino or pigmented, respectively, with respect to the values recorded from the non-operated contralateral eyes. These changes were maintained up to 12weeks post-ONT, demonstrating that the ERG–STR is a useful parameter to monitor RGC functionality in adult mice. [Copyright &y& Elsevier]

Abstract: Blue cone monochromacy (BCM) is an X-linked condition in which long- (L) and middle- (M) wavelength-sensitive cone function is absent. Due to the X-linked nature of the condition, female carriers are spared from a full manifestation of the associated defects but can show visual symptoms, including abnormal cone electroretinograms. Here we imaged the cone mosaic in four females carrying an L/M array with deletion of the locus control region, resulting in an absence of L/M opsin gene expression (effectively acting as a cone opsin knockout). On average, they had cone mosaics with reduced density and disrupted organization compared to normal trichromats. This suggests that the absence of opsin in a subset of cones results in their early degeneration, with X-inactivation the likely mechanism underlying phenotypic variability in BCM carriers. [ABSTRACT FROM AUTHOR]

Abstract: The spatial variation in central retinal function determined from mfERG was compared to co-localised measurements of cone density in two normal subjects. Individual cone cells in the parafoveal region of the retina were identified from 1°×1° images of the photoreceptor mosaic using a modified Heidelberg retina tomograph (HRT). The variation in cone density compared well with previous histology and retinal imaging studies and was strongly linearly correlated (r =0.98, p <0.001) with mfERG amplitude within the central retina. Retinal function determined from mfERG amplitude appears to directly reflect the density of the cone cells in this region. [Copyright &y& Elsevier]

Abstract: We investigated the color vision pattern in Cebus apella monkeys by means of electroretinogram measurements (ERG) and genetic analysis. Based on ERG we could discriminate among three types of dichromatic males. Among females, this classification is more complex and requires additional genetic analysis. We found five among 10 possible different phenotypes, two trichromats and three dichromats. We also found that Cebus present a new allele with spectral peak near 552nm, with the amino acid combination SFT at positions 180, 277 and 285 of the opsin gene, in addition to the previously described SYT, AFT and AFA alleles. [Copyright &y& Elsevier]

Abstract: Royal College of Surgeon (RCS) rats undergo retinal degeneration due to the inability of retinal pigment epithelial (RPE) cells to phagocytose shed outer segments. We explored the effect of introducing Schwann cells to the subretinal space of RCS rats (before the onset of retinal degeneration), by relying on electroretinogram (ERG) recordings and correlative retinal morphology. Scotopic ERGs recorded from cell-injected eyes showed preserved amplitudes of mixed a-wave b-wave, rod b-waves, and cone b-waves over controls (sham-injected eyes); photopic b-wave amplitudes and critical flicker fusion were also improved. Normal retinal morphology was found in areas of retinas that had received cell injections. Since Schwann cells have no phagocytic properties, their therapeutic effect is best explained through a paracrine mechanism (secretion of factors that ensure photoreceptor survival). [Copyright &y& Elsevier]

Abstract: The effects of pharmacological blockade of the proximal retinal activity by N-methyl-d-aspartate (MNDA) on the V–log I function of the ERG b- and d-waves were studied in dark and light adapted frog eyes. The effects of NMDA depended on the type of photoreceptor input. In rod-dominated ERG the b-wave amplitude was decreased by NMDA, but that of the d-wave remained unchanged. In cone-dominated ERG the b-wave amplitude was increased while the d-wave amplitude was decreased. Thus, the b/d amplitude ratio was decreased in rod-dominated, but increased in cone-dominated ERG. Our results imply that the b/d amplitude ratio would be changed in an opposite way in the rod- and cone-dominated ERG, when the function of the proximal retinal neurons is compromised. [Copyright &y& Elsevier]

Abstract: The electroretinogram (ERG) was obtained to contrast modulation (CM). This stimulus is a product of temporal modulation of the contrast of a spatial sinusoid at constant mean luminance. Mean contrast (10–40%), and modulation depth (25–1.0) were modulated at 7.5Hz to record the pattern electroretinogram (PERG). The spatial pattern was a foveally fixated grating pattern with sinusoidal luminance profile with spatial frequency of 4.6c/deg. CM resulted in significant first and second harmonic ERG responses. First harmonic amplitude increases then flattens as a function of mean contrast with ΔC =constant, while the second harmonic response remains unaffected by mean contrast. Apparently the first harmonic represents summed signals of local luminance responses arising from on and off neurons. Mean spatial contrast signals modulate preganglionic local luminance responses. [Copyright &y& Elsevier]

Abstract: We have characterized adaptive changes of inner retina function in response to sustained pattern stimulation in 32 normal subjects with an age range 23–77 years by measuring changes of the pattern electroretinogram (PERG) as a function of time. Contrast-reversal stimuli had square-wave profile in space and time, with peak spatial and temporal frequency and high contrast to maximize response amplitude. The PERG signal was sampled over 5min with a resolution of 15s. PERG signals were non-stationary, resulting in either progressive amplitude decline or even enhancement to a plateau, with a time course that could be well described by an exponential function with a time constant of 1–2min. Higher initial amplitudes were generally associated with amplitude decline, and lower initial amplitudes with enhancement. The delta amplitude (plateau minus initial) was a linear function of the initial amplitude. The magnitude of delta decreased with decreasing initial amplitude and inverted its sign for initial amplitudes about 1/3 lower than the maximum initial amplitude measured, but still about 3–4 times larger than the noise. Amplitude decline was generally associated with phase lag, whereas amplitude enhancement was associated with phase advance. Altogether, PERG generators appear to slowly adjust their gain in order to keep their sustained activity at an intermediate level that is rather independent of the level of activity at stimulus onset. This behavior is reminiscent of a buffering mechanism, where glial cells may play a primary role. An energy-budget model of neural–vascular–glial interaction is provided together with an equivalent electrical circuit that accounts for the results. [Copyright &y& Elsevier]

Abstract: The most widespread models to study blindness, rats and mice, have retinas containing less than 3% cones. The diurnal rodent Arvicanthis niloticus retina has around 35% cones. Using ERG recordings, we studied retina function in this species. Several features differed from that reported in rats and mice: (a) fivefold larger photopic a-wave amplitudes; (b) photopic hill effect in Nile grass rats only; and (c) flicker amplitude plateau between 5 to 35Hz with fusion beyond 60Hz in Nile grass rats only. We conclude that A. niloticus might complement rats and mice for studying retinal function and pathologies involving cones. [Copyright &y& Elsevier]

Abstract: The GUCA1A gene encodes a guanylate cyclase activating protein (GCAP1) that is involved in regulation of phototransduction in the vertebrate retina. We discovered a novel C312A transversion in exon 2 of the human GUCA1A gene, replacing Asn-104 (N104) in GCAP1 with Lys (K), in two affected members of a family with dominant cone dystrophy. The mutation N104K is located in the third EF-hand motif (EF3) shown previously to be instrumental in converting Ca2+-free GCAP1 to a GC inhibitor in the Ca2+-bound form. In one patient, rod ERGs were fairly stable over a 12-year-period whereas 30Hz flicker ERG and single-flash cone ERGs declined. In both patients, double-flash ERGs showed that rod recovery from an intense test flash was significantly delayed. The EC50 for GC stimulation shifted from ∼250nM in wild-type GCAP1 to ∼800nM in the GCAP1(N104K) mutant suggesting inability of the mutant to assume an inactive form under physiological conditions. The replacement of N104 by K in GCAP1 is the first naturally occurring mutation identified in the EF3 loop. The rod recovery delays observed in double-flash ERG of affected patients suggest a novel dominant-negative effect that slows GC stimulation. [Copyright &y& Elsevier]

Abstract: We characterize the dark-adapted photoresponses from mouse cones intact in the isolated retina, their virtually natural environment, by isolating pharmacologically the photoreceptor light responses from the electroretinogram (ERG). Due to the different photoresponse kinetics and sensitivity of rods and cones, the cone responses were readily attained by using a rod-saturating preflash. The stimulus wavelength (544nm) was chosen to selectively stimulate the green sensitive (“M”-)pigment. Obtained responses were monophasic, showing fast kinetics (mean t p =51ms) and low sensitivity (fractional single-photon response ca. 0.23%). The amplification coefficient of cones (4.6s−2) was very close to that of rods (5.6s−2), while the dominant time constant of recovery was clearly smaller for cones (33ms) than for rods (160ms). [Copyright &y& Elsevier]

Abstract: Focal macular cone electroretinograms (ERGs) and multifocal ERGs were recorded to study the macular function in patients with the complete-type of congenital stationary night blindness (cCSNB). The waveforms of the focal macular cone ERGs and the on- and off-responses of the multifocal ERGs in the cCSNB patients were similar to those recorded from monkey retinas treated with L-2 amino-4-phosphonobutyric acid (APB), suggesting that patients with cCSNB have a complete defect of the on-pathway even in the central retina. The results also demonstrated that there was a paradoxical positive response in the central retina of cCSNB patients, as compared to the negative full-field ERGs in the same subjects. [Copyright &y& Elsevier]

Abstract: The luminance–response function of the brief flash full-field photopic electroretinogram (ERG) rises to a peak before falling to a sub-maximal plateau – the ‘photopic hill’. The combination of on- and off-responses inherent in the brief flash photopic ERG suggests that this luminance–response function could be modelled by the sum of a Gaussian function and a logistic growth function. Photopic ERGs to a luminance series of brief flashes against three different background luminances recorded from seven healthy adults showed the characteristic ‘photopic hill’ function for b-wave amplitudes which were satisfactorily fitted with the sum of a Gaussian curve and a logistic growth curve. As background luminance increased, both components shifted to the right on the luminance axis. The Gaussian component increased in amplitude while the logistic growth function component decreased in amplitude. The luminance–response function of a complete congenital stationary night blindness patient had almost no logistic growth component. [Copyright &y& Elsevier]

Abstract: Shinoda and colleagues hypothesized that patients with cone dystrophy (CD) might suffer from a selective ON-system deficit, based on the local nature of the disease [Shinoda, K, Ohde, H, Inoue, R, Ishida, S, Mashima, Y, & Oguchi, Y (2002). ON-pathway disturbance in two siblings. Acta Ophthalmologica Scandinavica, 80, 219–223]. The purpose of the current study was to test this hypothesis by examining onset and offset responses as a function of eccentricity in a group of patients with CD using long-duration LED stimuli. Nine patients with CD participated in this study (mean age of 36.1 years and visual acuity ⩾20/200). For this study, the following measures were obtained: Humphrey threshold visual fields, standard multifocal ERGs (mfERGs) as well as mfERGs to long duration stimuli recorded using the Retiscan stimulator (Roland Instruments). This display contained 61 scaled hexagons and the LEDs were on for 100ms (180cd/m2) and off for 100ms. In addition, standard full-field photopic and flicker ERGs using Ganzfeld stimulation were obtained. For the control subjects, the onset responses were larger than the offset responses at all eccentricities; whereas for the patients, there was overlap between the amplitudes of the onset and offset responses. For the patients, the amplitude ratios (relative to the control data) indicated that the difference between the onset and offset responses was greatest for the central-most ring and this difference decreased with increasing eccentricity. For the onset responses, Humphrey thresholds and mfERG amplitudes, performance was poorest for the center ring and best for the most peripheral ring; for the offset responses, the opposite pattern of results was obtained. The differences in the pattern of results in the long duration mfERG data are consistent with a selective loss of the onset responses in our patient population. [Copyright &y& Elsevier]

Abstract: This study used the Matching Pursuit (MP) method, a time–frequency analysis, to identify and characterize oscillatory potentials (OPs) in the primate electroretinogram (ERG). When the slow-sequence mfERG from the macular region of the retina was matched with Gabor functions, OPs were identified in two distinct bands: a high-frequency band peaking around 150Hz that contributes to early OPs, and a low-frequency band peaking around 80Hz that contributes to both early and late OPs. Pharmacological blockade and experimental glaucoma studies showed that the high-frequency OPs depend upon sodium-dependent spiking activity of retinal ganglion cells, whereas the low-frequency OPs depend primarily upon non-spiking activity of amacrine cells, and more distal retinal activity. [Copyright &y& Elsevier]

Abstract: The contrast transfer function (CTF) of the pattern electroretinogram (PERG) depends on temporal frequency. For transient stimulation it is fully linear; at faster stimulation rates it becomes strongly non-linear with an accelerated shape. In this study we investigated a range of stimulus parameters with the aim of studying the influence of temporal and spatial frequencies, as well as contrast levels, on the CTF; effects were quantified via an “index of linearity” IL. Both reversal rate and check size influenced linearity (p <.001), examples: At a constant check size of 0.8°, 7.7rps: IL=1.0; 0.8°/24rps: IL=0.5; at a constant reversal rate of 19rps, IL was 0.5 for 0.8°, but rose to 0.8 both for 0.2° and 18°. The reason for this complex response surface remains a puzzle, it cannot be explained by varying parvo/magnocellular contributions, and its possible influences on recordings in patients merit further studies. [Copyright &y& Elsevier]

Melanopsin, a novel photopigment, has recently been localized to a population of retinal ganglion cells that display inherent photosensitivity. During continuous light and following light offset, primates are known to exhibit sustained pupilloconstriction responses that resemble closely the photoresponses of intrinsically-photoreceptive ganglion cells. We report that, in the behaving macaque, following pharmacological blockade of conventional photoreceptor signals, significant pupillary responses persist during continuous light and following light offset. These pupil responses display the unique spectral tuning, slow kinetics, and irradiance coding of the sustained, melanopsin-derived ganglion cell photoresponses. We extended our observations to humans by using the sustained pupil response following light offset to document the contribution of these novel ganglion cells to human pupillary responses. Our results indicate that the intrinsic photoresponses of intrinsically-photoreceptive retinal ganglion cells play an important role in the pupillary light reflex and are primarily responsible for the sustained pupilloconstriction that occurs following light offset. [ABSTRACT FROM AUTHOR]