Scotopic Flash Research Articles

Functional and morphological characterization of ocular phenotype in C3H/HeOuJ (Pde6b‐rd1) mice

Aims: C3H/HeOuJ mice (The Jackson Laboratory strain #000635) are homozygous for the retinal degeneration 1 mutation Pde6brd1 (rd1) causing severe visual deficits. The purpose of this study was to characterize the ocular phenotype of male and female mice with the rd1 mutation.Methods: The ophthalmic phenotype of mixed sex rd1 and aged‐matched C57BL/6J wild type (WT) mice (6 females and 4 males for rd1 group, 6 females and 5 males for WT group) was assessed at 3 months of age. Retinal photoreceptor function was quantified using scotopic flash electroretinography (fERG) with six different light intensities: 0.003, 0.01, 0.1, 1, 3 and 10 cd.s/m2 (Celeris, Diagnosys LLC). Retinal morphology was evaluated with spectral domain optical coherence tomography (SD‐OCT) (Envisu R2200, Bioptigen Inc./Leica Microsystems). Retinal thickness was measured from SD‐OCT images using a proprietary convolutional neural network. Optomotor reflex (OMR) was measured as photopic visual acuity and contrast sensitivity at 0.019 and 0.06 cyc/° (OptoDrum Plus, Striatech GmbH). Data were analysed by unpaired t‐test or Mann–Whitney test.Results: At 3 months of age, the rd1 mutation resulted in significant impairment of visual function in both male and female mice compared to age‐matched WT. fERG a and b‐wave amplitudes were decreased by 90% in both sexes (p < 0.001). The rd1 mice had only minimal visual acuity and contrast sensitivity assessed by OMR. Functional decline correlated with structural changes, notably the significant thinning of the retina (p < 0.001).Conclusions: The Pde6brd1 mutation resulted in severe functional and structural changes at 3 months of age. The mice presented significantly decreased but detectable fERG amplitudes, however, they had no functional vision as assessed by OMR. Combining fERG and OMR offers longitudinal in‐life quantification of retinal circuitry and functional vision with great utility for evaluation of novel therapeutic approaches for regeneration of the visual system.

Silicone oil insulation effects on flash electroretinogram and visual evoked potential in patients with retinal detachment

BackgroundSilicone oil is used as endotamponade following vitreoretinal surgery to maintain the retina reattached when indicated. This study investigates the hypothesis that silicone oil causes insulation effects on the retina by affecting its response to light. MethodsElectrophysiological responses to a flash stimulus were recorded using full-field electroretinography (ERG) and visual evoked potentials (VEP). Recordings were performed in 9 patients who underwent surgery for retinal detachment, before (1–2 days) and after (2–3 weeks) silicone oil removal (SOR) in both the study and the control eye. Flash ERG and VEP recordings were performed according to the ISCEV standard protocol. ResultsStatistically significant differences were found in the study eye in the amplitudes of the ERG responses and their corresponding ratios, i.e. the amplitude after SOR over the amplitude before SOR, in all conditions tested. No differences were observed in the control eye. The mean ratio of photopic ERG response was 3.4 ± 2.4 for the study and 1.0 ± 0.3 for the control eye (p<0.001). The mean ratio of ERG flicker response was 3.1 ± 2.4 and 1.0 ± 0.3, respectively (p = 0.003). Scotopic flash ERG ratio was 5.0 ± 4.4 for the study and 1.3 ± 0.6 for the control eye (p = 0.012). No differences were observed for the amplitude and latency of flash VEP response after SOR. ConclusionsSilicone oil causes a reduction in flash ERG responses; no effect was found on flash VEP responses. ERGs in eyes filled with silicone oil should not be considered representative of retinal functionality, in contrast to VEPs, which are not affected by silicone oil presence.

A Novel Autosomal Recessive Variant of the NRL Gene Causing Enhanced S-Cone Syndrome: A Morpho-Functional Analysis of Two Unrelated Pediatric Patients.

Enhanced S-cone syndrome (ESCS) is a rare autosomal recessive retinal degeneration mainly associated with pathogenic variations in the NR2E3 gene. Only a few pathogenic variations in the NRL gene associated with ESCS have been reported to date. Here, we describe the clinical and genetic findings of two unrelated pediatric patients with a novel frameshift homozygous variant in the NRL gene. Fundus examinations showed signs of peripheral degeneration in both patients, more severe in Proband 2, with relative sparing of the macular area. Spectral domain optical coherence tomography (SD-OCT) revealed a significant macular involvement with cysts in Proband 1, and minimal foveal alteration with peripheral retina involvement in Proband 2. Visual acuity was abnormal in both patients, but more severely affected in Proband 1 than Proband 2. The electroretinogram recordings showed reduced scotopic, mixed and single flash cone responses, with a typical supernormal S-cone response, meeting the criteria for a clinical diagnosis of ESCS in both patients. The present report expands the clinical and genetic spectrum of NRL-associated ESCS, and confirms the age-independent variability of phenotypic presentation already described in the NR2E3-associated ESCS.

Effects of chronic mild hyperoxia on retinal and choroidal blood flow and retinal function in the DBA/2J mouse model of glaucoma.

PurposeTo test the hypothesis that mild chronic hyperoxia treatment would improve retinal function despite a progressive decline in ocular blood flow in the DBA/2J mouse model of glaucoma.Materials and methodsDBA/2J mice were treated with chronic mild hyperoxia (30% O2) beginning at 4.5 months of age or were untreated by giving normal room air. Retinal and choroidal blood flow (RBF and ChBF, respectively) were measured at 4, 6, and 9 months of age by MRI. Blood flow was additionally measured under hypercapnia challenge (5% CO2 inhalation) to assess vascular reactivity. Intraocular pressure (IOP) was measured using a rebound tonometer at the same time points. Scotopic flash electroretinograms (ERGs) were recorded at 9 months of age.ResultsBoth ChBF and RBF were reduced and significantly affected by age (p < 0.01), but neither were significantly affected by O2-treatment (p > 0.05). ChBF significantly increased in response to hypercapnia (p < 0.01), which was also unaffected by O2-treatment. Significant effects of age (p < 0.001) and of the interaction of age with treatment (p = 0.028) were found on IOP. IOP significantly decreased in O2-treated mice at 6 months compared to 4 months of age (p < 0.001), while IOP trended to increase with age in untreated mice. The amplitude of the b-wave from ERG was significantly increased in O2-treated DBA/2J compared to the untreated mice (p = 0.012), while the a-wave and oscillatory potentials were not significantly affected (p > 0.05).ConclusionThis study investigated the effects of chronic mild hyperoxia on retinal function and on retinal and choroidal blood flow in a mouse model of glaucoma. Retinal function was improved in the O2-treated mice at late stage, despite a progressive decline of RBF and ChBF with age that was comparable to untreated mice.

Using the RETeval Device in Healthy Children to Establish Normative Electroretinogram Values.

To provide normative data of full-field electro-retinogram (ERG) responses in the pediatric population using the RETeval ERG device (LKC Technologies, Inc) in healthy children without evidence of retinal disease. This was a single-site cross-sectional study of healthy pediatric patients with normal dilated fundus examinations and no known retinal diseases. Participants were recruited to undergo dilated full-field ERG using the handheld RETeval device. The International Society for Clinical Electrophysiology of Vision 5-step protocol was used. Photopic and dark-adapted scotopic responses were recorded using skin electrodes. Main outcome measures were normative RETeval ERG values and correlation of age with measured ERG parameters. Thirty-eight eyes of 20 healthy patients (aged 4 to 17 years) were included in the study. Of the 20 normal patients, 9 were male and 11 were female. Normative mean, median, and range values were recorded for the measured full-field ERG parameters. Pearson correlation was moderately positive between age and oscillatory potential and scotopic dim flash amplitude (r = 0.59, P = .006 for both). A positive correlation was also found between age and cone a-wave implicit time (r = 0.67, P = .001). The handheld RETeval system is a useful tool for obtaining full-field ERGs in children without anesthesia. Moderate positive correlations were observed between age and oscillatory potential and scotopic dim flash amplitude. A strong correlation was found between age and cone a-wave implicit time. The current study provides a baseline of normative full-field ERG values in children. [J Pediatr Ophthalmol Strabismus. 2021;58(1):17-22.].

Electronegative electroretinograms in the United Arab Emirates

PURPOSE:An electronegative electroretinogram (ERG), defined as having a b:a wave ratio ≤1 in the scotopic flash ERG response, indicates relative inner retinal dysfunction. Causes vary depending upon the study population. In the Arabian Gulf, where inherited retinal disease is relatively prevalent, common diagnoses associated with electronegative ERGs have not been described. In this study, we report the frequency and causes of electronegative ERGs in a cohort of Emirati patients with inherited retinal disease.METHODS:A retrospective review was performed of all full-field ERGs done for Emirati patients in the Ocular Genetics Service of Cleveland Clinic Abu Dhabi from January 2017 to December 2019. Those who had an electronegative ERG in at least one eye were included in the study.RESULTS:Out of 137 patients, 9 probands (6.6%) had an electronegative ERG. The mean age at presentation was 24 years (range 5–48 years), and five patients (55.6%) were male. The final clinical diagnoses were congenital stationary night blindness (CSNB) (two TRPM1-related and one Oguchi disease), X-linked retinoschisis (XLRS) (one genetically confirmed and two not genetically tested), cone-rod dystrophy (one CRX-related and one not genetically tested), and enhanced S-cone syndrome (ESCS) (one NRL-related). The one patient who did not have bilateral electronegative ERGs was a male with XLRS whose fellow eye had an unrecordable ERG.CONCLUSIONS:In this series of Emirati patients, an electronegative ERG was most commonly associated with the inherited retinal diseases recessive CSNB and XLRS. An electronegative ERG was noted in a case of NRL-related ESCS.

Chapter 32 - Electroretinograms

The electroretinogram (ERG) is an electrical response of the retina to photic stimulation. A flash of light or bright appearance of a pattern elicits a biphasic negative/positive waveform. The a-wave originating in the receptor level of rods and cones is the initial large negative wave. The b-wave originating in the midretina is the following large positive component. Protocols include full-field and multifocal ERGs. Full-field ERGs are recorded if one is most interested in the global health of the retina, such as in retinitis pigmentosa. A limitation of the full-field ERG is that the recording is a massed potential from the whole retina. Multifocal ERGs can map small scotomas in the central 40+ degrees of visual field. Abbreviated methods are used with infants and in the operating room as part of an exam under anesthesia. The effects of toxic medications can be detected and quantified using ERGs. The ERG is useful to assess cases of retinal foreign bodies and trauma to estimate the extent of retinal dysfunction. Foreign bodies affect retinal function depending on the extent of trauma to the retina and the location and composition of the object. Systemic metabolic disorders usually reduce ERG b-wave amplitudes, particularly the scotopic dim flash ERGs.

Further Characterization of the Predominant Inner Retinal Degeneration of Aging Cln3 Δex7/8 Knock-In Mice.

Neuronal ceroid lipofuscinosis (NCL) is the most common group of neurogenetic storage diseases typically beginning in childhood. The juvenile form (JNCL), also known as Batten disease, is the most common form. Vision-related problems are often an early sign, appearing prior to motor and mental deficits. We have previously investigated disease progression with age in the Cln3 Δex7/8 KI mouse model for JNCL and showed a decline of visual acuity and a predominant decline of the inner retinal function in mice, similar to human disease. The aim of this study was to further characterize this degeneration by means of flicker ERGs. For the scotopic flicker ERG, we found a significantly lower magnitude for Cln3 Δex7/8 KI mice already at 6months of age for low stimulus frequencies, while the difference declines with increasing frequency. Under photopic conditions there was no magnitude difference at 6months, but a cumulative magnitude reduction with further aging. For both conditions the phases were similar for both groups. There was a similar magnitude reduction for the responses of both the slow and fast rod pathway in the 15 Hz experiments, and there were no differences in response phase. Low-frequency flicker responses seem to be sensitive to very early disease manifestations, and while the degeneration is associated with a reduction of predominating inner retinal responses in the scotopic flash ERG, this predominance seems not to be related to a selective involvement of the slow and fast rod pathways.

Test-retest reliability of scotopic full-field electroretinograms in rabbits.

To explore test-retest reliability of standard scotopic full-field ERG measurements in New Zealand White rabbits. TheERG is widely used for testing of retinal integrity after any ocular treatment. We here present detailed stimulus-response dependencies for single healthy and untreated animals, concentrating on test-retest reproducibility. Five New Zealand White rabbits (aged 8-10weeks, weight about 2.0-2.5kg) underwent binocular ERG measurements after intramuscular anesthesia and pharmaceutical pupillary dilatation at a baseline day and 10days later. Eleven scotopic flash strengths (0.0001-10cds/m2) were presented. Variability was quantified via the 95% limits of agreement (LOA). The a-waves displayed the typical monotonic sigmoid amplitude increase with flash strength, and the b-waves peaked at 0.01cds/m2, followed by a marked dip at 0.1-0.3cds/m2. LOA of both waves went through a maximum in the dip region. LOA divided by mean amplitudes (relative LOA) was fairly flat over flash strength, around 20% beyond the dip. Intraindividual interocular variability was markedly lower, around 10%. Scotopic ERGresponses in rabbits display a region of high variability at 0.1-0.3cds/m2; beyond that region the amplitude-LOA is 20%, the interocular LOA being half that value. The use of intraindividual control eyes for testing any toxicity of ocular agents thus appears markedly more sensitive. As a rule of thumb, we found the relative 95% LOA as 33% between individuals, 20% across sessions and 10% between eyes.

Asymmetrical Functional Deficits of ON and OFF Retinal Processing in the mdx3Cv Mouse Model of Duchenne Muscular Dystrophy.

The dystrophin mouse mutant mdx3Cv exhibits scotopic electroretinograpic (ERG) abnormalities, which resemble clinical changes observed in Duchenne muscular dystrophy (DMD) patients. In the present study, ERGs obtained from mdx3Cv and their wild-type littermates under scotopic, mesopic, and photopic conditions were analyzed to provide further insight on the affected retinal pathways, and to compare them with human data. Electroretinograms of mdx3Cv (n = 9) and age-matched C57BL/6J mice (n = 10) included the scotopic full-field flash (for outer retinal deficits in rod pathway), scotopic threshold response (for inner retinal integrity), photopic flash, sinusoidal flicker (for outer retinal deficits in cone pathway), mesopic rapid-on/-off sawtooth flicker, and photopic long-duration flash measurements (for separate ON-/OFF-responses under different conditions). The mdx3Cv mice exhibited diminished and delayed scotopic and photopic ERGs, particularly in their b-wave and oscillatory potentials. Interestingly, homologues to the a- and b-wave of the mesopic ON-response were affected in their peak/trough times but not in their amplitude, whereas changes to both features were uncovered for photopic ON-response and sinusoidal flicker. Mesopic and photopic OFF-components were within the norm. Abnormal scotopic and photopic flash ERGs were observed in mdx3Cv, which corroborate with deficits that are likely restricted to the level of photoreceptor-to-bipolar cell transmission. Further overlaps between mdx3Cv mice and DMD patients exist, including asymmetrical ON versus OFF ERG alterations under mesopic versus photopic vision. In mice, ON-pathway function is compromised, whereas the OFF-pathway is spared.

Systematic assessment of clinical methods to diagnose and monitor diabetic retinal neuropathy

PurposeBackground and Purpose: Diabetic retinal neuropathy refers to retinal neural tissue damage occurring before diabetic retinopathy and fulfils many of the criteria for causality for the subsequent vasculopathy. Developing reliable means of measuring neuronal damage in diabetes may be important in efforts to prevent retinopathy. This study aimed to systematically assess current clinical measurements of diabetic retinal neuropathy.MethodsA systematic search of the medical literature since 1984 was performed on PUBMED and EMBASE and the evidence supporting each identified method as an indicator for clinically important diabetic retinal neuropathy was graded relatively as strong, medium or weak according to criteria assessing its relationship to subsequent diabetic retinopathy, quality of supporting studies and published reproducibility.ResultsThe systematic search yielded 6,421 results. Subsequent assessment by two independent investigators identified 601 multiple subject studies in humans assessing clinical aspects of retinal structure, function or psychophysics in the pre‐diabetic retina. Clinical methods assessed as being supported by relatively ‘strong’ evidence included FM‐100 hue colour vision changes, flash ERG b‐wave latency, flash multifocal b‐wave latency, scotopic flash ERG oscillatory potential amplitude and contrast sensitivity.ConclusionsConclusions/Discussions: The results showed moderately poor quality of extant evidence and indicate the best clinical methods for assessing diabetic retinal neuropathy remain to be confirmed. This is the first systematic assessment of the medical literature aiming to assess the breadth and validity of these methods and represents an early step in identifying and developing endpoints for use in trials designed to identify at risk patients or prevent diabetic retinopathy.

In vitro and in vivo neuroprotective effects of cJun N-terminal kinase inhibitors on retinal ganglion cells.

BackgroundThe c-Jun N-terminal kinase (JNK) signaling pathway plays an important role in neuronal pathophysiology. Using JNK inhibitors, we examined involvement of the JNK pathway in cultured rat retinal ganglion cell (RGC) death and in mouse retinal ischemia/reperfusion (I/R) injury of the visual axis. The in vitro effects of JNK inhibitors were evaluated in cultured adult rat retinal cells enriched in RGCs. Retinal I/R was induced in C57BL/6J mice through elevation of intraocular pressure to 120 mmHg for 60 min followed by reperfusion. SP600125 was administered intraperitoneally once daily for 28 days. Phosphorylation of JNK and c-Jun in the retina was examined by immunoblotting and immunohistochemistry. The thickness of retinal layers and cell numbers in the ganglion cell layer (GCL) were examined using H&E stained retinal cross sections and spectral domain optical coherence tomography (SD-OCT). Retinal function was measured by scotopic flash electroretinography (ERG). Volumetric measurement of the superior colliculus (SC) as well as VGLUT2 and PSD95 expression were studied.ResultsJNK inhibitors SP600125 and TAT-JNK-III, dose-dependently and significantly (p < 0.05) protected against glutamate excitotoxicity and trophic factor withdrawal induced RGC death in culture. In the I/R model, phosphorylation of JNK (pJNK) in the retina was significantly (p < 0.05) increased after injury. I/R injury significantly (p < 0.05) decreased the thickness of retinal layers, including the whole retina, inner plexiform layer, and inner nuclear layer and cell numbers in the GCL. Administration of SP600125 for 28 days protected against all these degenerative morphological changes (p < 0.05). In addition, SP600125 significantly (p < 0.05) protected against I/R-induced reduction in scotopic ERG b-wave amplitude at 3, 7, 14, 21 and 28 days after injury. SP600125 also protected against the I/R-induced losses in volume and levels of synaptic markers in the SC. Moreover, the protective effects of SP600125 in the retina and SC were also detected even with only 7 days (Days 1–7 after I/R) of SP600125 treatment.ConclusionsOur results demonstrate the important role the JNK pathway plays in retinal degeneration in both in vitro and in vivo models and suggest that JNK inhibitors may be a useful therapeutic strategy for neuroprotection of RGCs in the retina.Electronic supplementary materialThe online version of this article (doi:10.1186/s13024-016-0093-4) contains supplementary material, which is available to authorized users.

Short Communication: Contribution of Voltage-Gated Sodium Channels to the Rabbit Cone Electroretinograms

ABSTRACTPurpose: Although the rabbit eye is of a similar size to the human eye, our limited understanding of the differences in retinal physiology to other species hinders its use in retinal research. The role of voltage-gated sodium channels (Nav) in the propagation of excitatory potentials along bipolar cells remains unclear, as conflicting data have been reported in the rabbit. The present study assesses the relative contributions of Nav to the scotopic and photopic flash ERGs as well as the wavelength-dependence of Nav blockade on the rabbit flicker ERG.Materials and Methods: Tetrodotoxin (TTX, 1 μM) was injected into the vitreous cavity of Chinchilla bastard rabbits. Scotopic ERGs were evoked by white flashes ranging from 10−5 to 10 cds m−2, photopic ERGs on a background of 25 cdm−2 using flash intensities of 0.032–25 cds m−2. Flicker ERGs (3–50 Hz) were elicited by blue, green and yellow stimuli at 2.34 cds m−2 on a white background of 30 cdm−2.Results: The a- and b-waves of the scotopic ERG were unaffected by intravitreal injection of the Nav blocker TTX. In contrast, the b-wave, but not the a-wave, of the photopic ERG was selectively blocked by TTX. The reduction by TTX of the flicker ERG was greater for blue than for green and yellow stimuli.Discussion: The data suggest that Nav selectively contribute to the generation of the photopic b-wave in the rabbit, indicating that they play an important role in the propagation of excitatory signals on bipolar cells in the cone, but not rod pathways. Importantly, the present study resolves conflicting previous reports into the role of Nav in the retinal function of the rabbit.

Early adaptive response of the retina to a pro-diabetogenic diet: Impairment of cone response and gene expression changes in high-fructose fed rats

The lack of plasticity of neurons to respond to dietary changes, such as high fat and high fructose diets, by modulating gene and protein expression has been associated with functional and behavioral impairments that can have detrimental consequences. The inhibition of high fat-induced rewiring of hypothalamic neurons induced obesity. Feeding rodents with high fructose is a recognized and widely used model to trigger obesity and metabolic syndrome. However the adaptive response of the retina to short term feeding with high fructose is poorly documented. We therefore aimed to characterize both the functional and gene expression changes in the neurosensory retina of Brown Norway rats fed during 3 and 8 days with a 60%-rich fructose diet (n = 16 per diet and per time point). Glucose, insulin, leptin, triacylglycerols, total cholesterol, HDL-cholesterol, LDL-cholesterol and fructosamine were quantified in plasma (n = 8 in each group). Functionality of the inner retina was studied using scotopic single flash electroretinography (n = 8 in each group) and the individual response of rod and cone photoreceptors was determined using 8.02 Hz Flicker electroretinography (n = 8 in each group). Analysis of gene expression in the neurosensory retina was performed by Affymetrix genechips, and confirmed by RT-qPCR (n = 6 in each group). Elevated glycemia (+13%), insulinemia (+83%), and leptinemia (+172%) was observed after 8 days of fructose feeding. The cone photoreceptor response was altered at day 8 in high fructose fed rats (Δ = 0.5 log unit of light stimulus intensity). Affymetrix analysis of gene expression highlighted significant modulation of the pathways of eIF2 signaling and endoplasmic reticulum stress, regulation of eIF4 and p70S6K signaling, as well as mTOR signaling and mitochondrial dysfunction. RT-qPCR analysis confirmed the down regulation of Crystallins, Npy, Nid1 and Optc genes after 3 days of fructose feeding, and up regulation of End2. Meanwhile, a trend towards an increased expression of αA- and αB-crystallin proteins was observed at day 8. Our results are consistent with early alterations of the functioning and gene expression in the retina in a pro diabetogenic environment.

Acetylcholine induces GABA release onto rod bipolar cells through heteromeric nicotinic receptors expressed in A17 amacrine cells.

Acetylcholine (ACh) is a major retinal neurotransmitter that modulates visual processing through a large repertoire of cholinergic receptors expressed on different retinal cell types. ACh is released from starburst amacrine cells (SACs) under scotopic conditions, but its effects on cells of the rod pathway have not been investigated. Using whole-cell patch clamp recordings in slices of rat retina, we found that ACh application triggers GABA release onto rod bipolar (RB) cells. GABA was released from A17 amacrine cells and activated postsynaptic GABAA and GABAC receptors in RB cells. The sensitivity of ACh-induced currents to nicotinic ACh receptor (nAChR) antagonists (TMPH ~ mecamylamine > erysodine > DhβE > MLA) together with the differential potency of specific agonists to mimic ACh responses (cytisine >> RJR2403 ~ choline), suggest that A17 cells express heteromeric nAChRs containing the β4 subunit. Activation of nAChRs induced GABA release after Ca2+ accumulation in A17 cell dendrites and varicosities mediated by L-type voltage-gated calcium channels (VGCCs) and intracellular Ca2+ stores. Inhibition of acetylcholinesterase depolarized A17 cells and increased spontaneous inhibitory postsynaptic currents in RB cells, indicating that endogenous ACh enhances GABAergic inhibition of RB cells. Moreover, injection of neostigmine or cytisine reduced the b-wave of the scotopic flash electroretinogram (ERG), suggesting that cholinergic modulation of GABA release controls RB cell activity in vivo. These results describe a novel regulatory mechanism of RB cell inhibition and complement our understanding of the neuromodulatory control of retinal signal processing.

Flash electroretinography in the bald eagle (Haliaeetus leucocephalus).

Photopic and scotopic flash electroretinograms (fERGs) were done on 12 adult captive anesthetized bald eagles (Haliaeetus leucocephalus) following a complete ophthalmic exam. The b-wave amplitude in the bald eagle exceeds that seen in other species when using a similar protocol. This data may be used clinically as a reference for bald eagles undergoing fERG evaluation for retinal disease or as a preoperative screening tool before phacoemulsification.

Transplantation of Photoreceptors Derived From Human Müller Glia Restore Rod Function in the P23H Rat

Müller glia possess stem cell characteristics that have been recognized to be responsible for the regeneration of injured retina in fish and amphibians. Although these cells are present in the adult human eye, they are not known to regenerate human retina in vivo. Human Müller glia with stem cell characteristics (hMSCs) can acquire phenotypic and genotypic characteristics of rod photoreceptors in vitro, suggesting that they may have potential for use in transplantation strategies to treat human photoreceptor degenerations. Much work has been undertaken in rodents using various sources of allogeneic stem cells to restore photoreceptor function, but the effect of human Müller glia-derived photoreceptors in the restoration of rod photoreceptor function has not been investigated. This study aimed to differentiate hMSCs into photoreceptor cells by stimulation with growth and differentiation factors in vitro to upregulate gene and protein expression of CRX, NR2E3, and rhodopsin and various phototransduction markers associated with rod photoreceptor development and function and to examine the effect of subretinal transplantation of these cells into the P23H rat, a model of primary photoreceptor degeneration. Following transplantation, hMSC-derived photoreceptor cells migrated and integrated into the outer nuclear layer of the degenerated retinas and led to significant improvement in rod photoreceptor function as shown by an increase in a-wave amplitude and slope using scotopic flash electroretinography. These observations suggest that hMSCs can be regarded as a cell source for development of cell-replacement therapies to treat human photoreceptor degenerations and may also offer potential for the development of autologous transplantation.

Slight Alteration of the Electroretinogram in Mice Lacking Dystrophin Dp71

Aim: Most Duchenne muscular dystrophy patients and the mdx^Cv3 mouse strain, lacking expression of both dystrophins Dp260 and Dp71, show a high attenuation of the dark-adapted electroretinogram (ERG) b-wave amplitude, whereas mice lacking the expression of Dp260 show normal b-wave amplitude. Here, we completed our assessment of whether the sole absence of Dp71 affects the ERG. Methods: Ganzfeld ERGs were performed on dark-adapted Dp71-null mice and littermates. Scotopic flash ERGs were recorded at light intensities from 3.10−⁵ to 1 cd.s/m². Oscillatory potentials (OPs) were extracted at 1 cd.s/m². Photopic flash ERGs were recorded at 10 cd.s/m² after light adaptation. Results: Dp71-null mice showed a slight but significant reduction in b-wave amplitudes, normal a-wave amplitudes and nonaffected implicit times of the scotopic ERGs. No changes were observed in the amplitudes and implicit times of the OPs and the photopic ERGs. Conclusions: Our results demonstrate that together both Dp71 and Dp260 are required for the generation of the ERG b-wave in mice.

Non-selectivity of ERG reductions in eyes treated for retinoblastoma

We have monitored retinal function in patients treated for retinoblastoma (primarily, but not exclusively by intra-arterial chemotherapy infusion) by electroretinography (ERG) recordings for the past 7years. We here present data from 599 ERG studies of 108 patients, in which a complete ERG protocol including both photopic and scotopic recordings was performed, in justification of our frequent practice of reporting primarily 30-Hz photopic flicker amplitude data. Patients referred for treatment of retinoblastoma underwent ERG recordings during examination under anesthesia whenever possible: at baseline and following most treatment sessions. Correlations were calculated for the complete datasets between the four primary amplitude response parameters: photopic single flash b-wave, photopic 30-Hz flicker peak-to-trough, scotopic rod-isolating b-wave, and scotopic maximal flash b-wave. Using our adaptation of the International Society for Clinical Electrophysiology of Vision-recommended standard ERG protocol, ERG responses of eyes of patients with untreated retinoblastoma or following traditional or intra-arterial treatment for retinoblastoma show very high correlations between 30-Hz flicker amplitude responses and three other standard photopic and scotopic ERG response amplitudes. Reductions in ERG amplitudes seen in these eyes following treatment show no significant difference between retinal dysfunction estimated using rod- or cone-dominated responses. These observations support the use of photopic response amplitudes (especially in response to 30-Hz flicker) as the primary ERG outcome measure in studies of treated and untreated eyes with retinoblastoma when more complete ERG protocols may be impractical.

Functional protective effects of long-term memantine treatment in the DBA/2J mouse

To analyse the effects of long-term memantine treatment on the retinal physiology and morphology of DBA/2J mice. DBA/2J (D2J) mice received i.p. injections of the NMDA receptor antagonist memantine, which protects neurons from abnormally elevated glutamate levels, twice a day over a period of 7 months. At the age of 2, 6 and 10 months, the intraocular pressure (IOP) and electroretinograms (ERGs) were measured in all treated D2J mice, in untreated D2J controls and in C57Bl/6 (B6) wild-type mice. After the last measurement at the age of 10 months, the mice were killed and the retinae and the optic nerves were analysed morphologically. The IOP increased with age in both D2J and B6 mice with a larger increase in the D2J strain. IOPs were not influenced by memantine treatment. The response amplitude of the scotopic flash ERG decreased with age in the D2J strain. This amplitude decrease, particularly that of the b-wave, was smaller in treated D2J mice. The retinae of treated D2J mice exhibited less peripheral degeneration of cone photoreceptors, and optic nerve neuropathy was less frequent. Application of the NMDA receptor antagonist memantine diminished retinal neurodegeneration in the D2J mice and had a protective effect on the b-wave amplitude of the scotopic flash ERG. This protection may occur secondarily as memantine primarily acts on retinal ganglion cells.