Visual electrophysiology and "the potential of the potentials"

Visual electrophysiology affords direct, quantitative, objective assessment of visual pathway function at different levels, and thus yields information complementary to, and not necessarily obtainable from, imaging or psychophysical testing. The tests available, and their indications, have evolved, with many advances, both in technology and in our understanding of the neural basis of the waveforms, now facilitating more precise evaluation of physiology and pathophysiology. After summarising the visual pathway and current standard clinical testing methods, this review discusses, non-exhaustively, several developments, focusing particularly on human electroretinogram recordings. These include new devices (portable, non-mydiatric, multimodal), novel testing protocols (including those aiming to separate rod-driven and cone-driven responses, and to monitor retinal adaptation), and developments in methods of analysis, including use of modelling and machine learning. It is likely that several tests will become more accessible and useful in both clinical and research settings. In future, these methods will further aid our understanding of common and rare eye disease, will help in assessing novel therapies, and will potentially yield information relevant to neurological and neuro-psychiatric conditions

Human retinal dark adaptation tracked in vivo with the electroretinogram: insights into processes underlying recovery of cone- and rod-mediated vision

The substantial time taken for regaining visual sensitivity (dark adaptation) following bleaching exposures has been investigated for over a century. Psychophysical studies yielded the classic biphasic curve representing recovery of cone-driven and rod-driven vision. The electroretinogram (ERG) permits direct assessment of recovery at the level of the retina (photoreceptors, bipolar cells), with the first report over 70 years ago. Over the last two decades, ERG studies of dark adaptation have generated insights into underlying physiological processes. After large bleaches, rod photoreceptor circulating current, estimated from the rod-isolated bright-flash ERG a-wave, takes 30 min to recover, indicating that products of bleaching, thought to be free opsin (unbound to 11-cis-retinal), continue to activate phototransduction, shutting off rod circulating current. In contrast, cone current, assessed with cone-driven bright-flash ERG a-waves, recovers within 100 ms following similar exposures, suggesting that free opsin is less able to shut off cone current. The cone-driven dim-flash a-wave can be used to track recovery of cone photopigment, showing regeneration is 'rate-limited' rather than first order. Recoveries of the dim-flash ERG b-wave are consistent also with rate-limited rod photopigment regeneration (where free opsin, desensitising the visual system as an 'equivalent background', is removed by rate-limited delivery of 11-cis-retinal). These findings agree with psychophysical and retinal densitometry studies, although there are unexplained points of divergence. Post-bleach ERG recovery has been explored in age-related macular degeneration and in trials of visual cycle inhibitors for retinal diseases. ERG tracking of dark adaptation may prove useful in future clinical contexts

Negative electroretinograms: genetic and acquired causes, diagnostic approaches and physiological insights.

Funder: Moorfields Eye CharityThe dark-adapted human electroretinogram (ERG) response to a standard bright flash includes a negative-going a-wave followed by a positive-going b-wave that crosses the baseline. An electronegative waveform (or negative ERG) results when the b-wave is selectively reduced such that the ERG fails to cross the baseline following the a-wave. In the context of a normally sized a-wave, it indicates a site of retinal dysfunction occurring after phototransduction (commonly at the photoreceptor to bipolar cell synapse). This is an important finding. In genetic disease, the pattern of ERG abnormality can point to variants in a small group of genes (frequently those associated with congenital stationary night blindness and X-linked retinoschisis, but negative ERGs can also be seen in other conditions including syndromic disease). In acquired disease, there are numerous causes, but specific features may point to melanoma-associated retinopathy (MAR). In some cases, the visual symptoms precede the diagnosis of the melanoma and so the ERG findings can initiate investigations facilitating early detection and treatment. Negative ERGs can occur in other paraneoplastic conditions, and in a range of other diseases. This review will outline the physiological basis for the negative ERG, report prevalences in the literature from different cohorts, discuss the range of causes, displaying examples of a number of ERG phenotypes, highlight features of a clinical approach to patients, and briefly discuss further insights relating to current flows shaping the a-wave trough and from single-cell transcriptome analysis

Retinal Ganglion Cells-Diversity of Cell Types and Clinical Relevance.

Retinal ganglion cells (RGCs) are the bridging neurons that connect the retinal input to the visual processing centres within the central nervous system. There is a remarkable diversity of RGCs and the various subtypes have unique morphological features, distinct functions, and characteristic pathways linking the inner retina to the relevant brain areas. A number of psychophysical and electrophysiological tests have been refined to investigate this large and varied population of RGCs. Technological advances, such as high-resolution optical coherence tomography imaging, have provided additional tools to define the pattern of RGC involvement and the chronological sequence of events in both inherited and acquired optic neuropathies. The mechanistic insights gained from these studies, in particular the selective vulnerability and relative resilience of particular RGC subtypes, are of fundamental importance as they are directly relevant to the development of targeted therapies for these invariably progressive blinding diseases. This review provides a comprehensive description of the various types of RGCs, the developments in proposed methods of classification, and the current gaps in our knowledge of how these RGCs are differentially affected depending on the underlying aetiology. The synthesis of the current body of knowledge on the diversity of RGCs and the pathways that are potentially amenable to therapeutic modulation will hopefully lead to much needed effective treatments for patients with optic neuropathies

The Diagnostic Accuracy of Photopic Negative Responses Evoked by Broadband and Chromatic Stimuli in a Clinically Heterogeneous Population

Purpose: To compare the diagnostic accuracy of the photopic negative response (PhNR) elicited by red-blue (RB) and white-white (WW) stimuli, for detection of retinal ganglion cell (RGC) dysfunction in a heterogeneous clinical cohort. Methods: Adults referred for electrophysiological investigations were recruited consecutively for this single-centre, prospective, paired diagnostic accuracy study. PhNRs were recorded to red flashes (1.5 cd·s·m−2) on a blue background (10 cd·m−2) and to white flashes on a white background (the latter being the ISCEV standard LA 3 stimulus). PhNR results were compared with a reference test battery assessing RGC/optic nerve structure and function including optical coherence tomography (OCT) retinal nerve fibre layer thickness and mean RGC volume measurements, fundus photography, pattern electroretinography and visual evoked potentials. Primary outcome measures were differences in sensitivity and specificity of the two PhNR methods. Results: Two hundred and forty-three participants were initially enrolled, with 200 (median age 54; range 18–95; female 65%) meeting inclusion criteria. Sensitivity was 53% (95% confidence intervals [CI] 39% to 68%) and 62% (95% CI 48% to 76%), for WW and RB PhNRs, respectively. Specificity was 80% (95% CI 74% to 86%) and 78% (95% CI 72% to 85%), respectively. There was a statistically significant difference between sensitivities (p = 0.046) but not specificities (p = 0.08) of the two methods. Receiver operator characteristic (ROC) area under the curve (AUC) values were 0.73 for WW and 0.74 for RB PhNRs. Conclusion: PhNRs to red flashes on a blue background may be more sensitive than white-on-white stimuli, but there is no significant difference between specificities. This study highlights the value and potential convenience of using white-on-white stimuli, already used widely for routine ERG assessment

Prognostication in Stargardt disease using Fundus Autofluorescence: Improving Patient Care

PURPOSE: To explore fundus autofluorescence (FAF) imaging as an alternative to electroretinogram (ERG), as a non-invasive, quick, and readily interpretable method to predict disease progression in Stargardt disease (STGD). DESIGN: Retrospective case series of patients who attended Moorfields Eye Hospital (London, UK). SUBJECTS: Patients with STGD who met the following criteria were included: (i) biallelic disease-causing variants in ABCA4, (ii) ERG testing performed inhouse with an unequivocal ERG group classification, and (iii) ultra-widefield (UWF) FAF imaging performed up to 2 years before or after the ERG. METHODS: Patients were divided into three ERG groups based on retinal function and three FAF groups according to the extent of the hypoautofluorescence and their retinal background appearance. FAF imaging of 30 and 55° were also subsequently reviewed. MAIN OUTCOME MEASURES: ERG/FAF concordance and its association with baseline visual acuity and genetics. RESULTS: 234 patients were included in the cohort. 170 patients (73%) had the same ERG and FAF group, 33 (14%) had a milder FAF than ERG group, and 31 (13%) had a more severe FAF than ERG group. Children under the age of 10 (n=23) had the lowest ERG/FAF concordance, 57% (9 out of the 10 with discordant ERG/FAF had milder FAF than ERG), and adults with adult onset had the highest (80%). Missense genotypes were more commonly seen in the mildest phenotypes. In 97% and 98% of the cases, respectively, 30° and 55° FAF imaging matched with the group defined by UWF FAF. CONCLUSIONS: We demonstrate that FAF imaging is an effective modality to determine the extent of retinal involvement and thereby inform prognostication, by comparing FAF to the current gold standard of ERG testing to determine retinal involvement and thereby prognosis. In 80% of patients in our large molecularly proven cohort we were able to predict if the disease was confined to the macula or also affected the peripheral retina. Children assessed at a young age, with at least one null variant, early disease onset, and/or poor initial VA may have wider retinal involvement than predicted by FAF alone and/or progress to a more severe FAF phenotype over time

Cone-driven strong flash electroretinograms in healthy adults: Prevalence of negative waveforms

Purpose Both rod and cone-driven signals contribute to the electroretinogram (ERG) elicited by a standard strong flash in the dark. Negative ERGs usually reflect inner retinal dysfunction. However, in diseases where rod photoreceptor function is selectively lost, a negative waveform might represent the response of the dark-adapted cone system. To investigate the dark-adapted cone-driven waveform in healthy individuals, we delivered flashes on a dim blue background, designed to saturate the rods, but minimally adapt the cones. Methods ERGs were recorded, using conductive fibre electrodes, in adults from the TwinsUK cohort. Responses to 13 cd m−2 s white xenon flashes (similar to the standard DA 10 flash), delivered on a blue background, were analysed. Photopic and scotopic strengths of the background were 1.3 and 30 cd m−2, respectively; through a dilated pupil, this is expected to largely saturate the rods, but adapt the cones much less than the standard ISCEV background. Results Mean (SD) participant age was 62.5 (11.3) years (93% female). ERGs from 203 right and 204 left eyes were included, with mean (SD) b/a ratios of 1.22 (0.28) and 1.18 (0.28), respectively (medians, 1.19 and 1.17). Proportions with negative waveforms were 23 and 26%, respectively. Right and left eye b/a ratios were strongly correlated (correlation coefficient 0.74, p < 0.0001). We found no significant correlation of b/a ratio with age. Conclusions Over 20% of eyes showed b/a ratios less than 1, consistent with the notion that dark-adapted cone-driven responses to standard bright flashes can have negative waveforms. The majority had ratios greater than 1. Thus, whilst selective loss of rod function can yield a negative waveform (with reduced a-wave) in some, our findings also suggest that loss of rod function can occur without necessarily yielding a negative ERG. One potential limitation is possible mild cone system adaptation by the background