Photoreceptor Research Articles

Retinal Degeneration Caused by Rod-Specific Dhdds Ablation Occurs without Concomitant Inhibition of Protein N-Glycosylation.

SummaryDehydrodolichyl diphosphate synthase (DHDDS) catalyzes the committed step in dolichol synthesis. Recessive mutations in DHDDS cause retinitis pigmentosa (RP59), resulting in blindness. We hypothesized that rod photoreceptor-specific ablation of Dhdds would cause retinal degeneration due to diminished dolichol-dependent protein N-glycosylation. Dhddsflx/flx mice were crossed with rod-specific Cre recombinase-expressing (Rho-iCre75) mice to generate rod-specific Dhdds knockout mice (Dhddsflx/flx iCre+). In vivo morphological and electrophysiological evaluation of Dhddsflx/flx iCre+ retinas revealed mild retinal dysfunction at postnatal (PN) 4 weeks, compared with age-matched controls; however, rapid photoreceptor degeneration ensued, resulting in almost complete loss of rods and cones by PN 6 weeks. Retina dolichol levels were markedly decreased by PN 4 weeks in Dhddsflx/flx iCre+ mice, relative to controls; despite this, N-glycosylation of retinal proteins, including opsin (the dominant rod-specific glycoprotein), persisted in Dhddsflx/flx iCre+ mice. These findings challenge the conventional mechanistic view of RP59 as a congenital disorder of glycosylation.

Segmented Macular Layer Volumes from Spectral Domain Optical Coherence Tomography in 184 Adult Twins: Associations With Age and Heritability.

PurposeTo investigate segmented macular layer volumes from a healthy adult twin cohort (TwinsUK), exploring changes with age and heritability.MethodsMacular spectral domain optical coherence tomography images were acquired from monozygotic (MZ) and dizygotic (DZ) twins in a cross-sectional study. The following layer volumes were derived for circles of 3 and 6 mm diameter around the foveal center, using automated segmentation software: retinal nerve fiber layer (RNFL), ganglion cell–inner plexiform layer (GCIPL), inner nuclear layer (INL), outer plexiform layer (OPL), outer nuclear layer (ONL), photoreceptors (PR), retinal pigment epithelium (RPE), and total retinal volume (TRV). Correlation coefficients (intereye; age; intrapair for MZ and DZ pairs) were quantified; heritability was estimated using structural equation modeling.ResultsScans from 184 participants were included. Intereye correlation was highest for TRV and GCIPL. Negative correlations with age (for 3- or 6-mm areas, or both) were observed for TRV, RNFL, GCIPL, and INL. Positive correlations were observed for PR, RPE, and OPL. For all layers, intrapair correlation was greater for MZ than DZ pairs. Heritability estimates were highest (>80%) for TRV and GCIPL volume, and lowest for RPE volume.ConclusionsAlthough TRV was negatively correlated with age, all layers did not show negative correlation. Some inner layers thinned with age, whereas some outer volumes increased (not the ONL). Reduced RPE phagocytic function with age and remodeling in the OPL could be contributing factors. Heritability estimates were highest for inner retinal layers (particularly GCIPL), and lowest for RPE volume.

Altered photoreceptor metabolism in mouse causes late stage age-related macular degeneration-like pathologies

Age-related macular degeneration (AMD) is the leading cause of blindness in the elderly. While the histopathology of the different disease stages is well characterized, the cause underlying the progression, from the early drusen stage to the advanced macular degeneration stage that leads to blindness, remains unknown. Here, we show that photoreceptors (PRs) of diseased individuals display increased expression of two key glycolytic genes, suggestive of a glucose shortage during disease. Mimicking aspects of this metabolic profile in PRs of wild-type mice by activation of the mammalian target of rapamycin complex 1 (mTORC1) caused early drusen-like pathologies, as well as advanced AMD-like pathologies. Mice with activated mTORC1 in PRs also displayed other early disease features, such as a delay in photoreceptor outer segment (POS) clearance and accumulation of lipofuscin in the retinal-pigmented epithelium (RPE) and of lipoproteins at the Bruch's membrane (BrM), as well as changes in complement accumulation. Interestingly, formation of drusen-like deposits was dependent on activation of mTORC1 in cones. Both major types of advanced AMD pathologies, including geographic atrophy (GA) and neovascular pathologies, were also seen. Finally, activated mTORC1 in PRs resulted in a threefold reduction in di-docosahexaenoic acid (DHA)-containing phospholipid species. Feeding mice a DHA-enriched diet alleviated most pathologies. The data recapitulate many aspects of the human disease, suggesting that metabolic adaptations in photoreceptors could contribute to disease progression in AMD. Identifying the changes downstream of mTORC1 that lead to advanced pathologies in mouse might present new opportunities to study the role of PRs in AMD pathogenesis.

Ferrous but not ferric iron sulfate kills photoreceptors and induces photoreceptor-dependent RPE autofluorescence

Iron has been implicated in the pathogenesis of retinal degenerative diseases, including ocular siderosis. However, the mechanisms of iron-induced retinal toxicity are incompletely understood. Previous work shows that intravitreal injection of Fe2+ leads to photoreceptor (PR) oxidative stress, resulting in PR death within 14 days, and cones are more susceptible than rods to iron-induced oxidative damage. In order to further investigate the mechanism of intravitreal iron-induced retinal toxicity and shed light on mechanisms of iron-induced retinopathy in other mouse models, Fe2+, Fe3+, or saline were injected into the vitreous of adult wild-type mice. Pre-treatment with Ferrostatin-1 was used to investigate whether iron-induced retinal toxicity resulted from ferroptosis. Color and autofluorescence in vivo retinal imaging and optical coherence tomography were performed on day 2 and day 7 post-injection. Eyes were collected for quantitative PCR and Western analysis on day 1 and for immunofluorescence on both day 2 and 7. In vivo imaging and immunofluorescence revealed that Fe2+, but not Fe3+, induced PR oxidative damage and autofluorescence on day 2, resulting in PR death and retinal pigment epithelial cell (RPE) autofluorescence on day 7. Quantitative PCR and Western analysis on day 1 indicated that both Fe2+ and Fe3+ induced iron accumulation in the retina. However, only Fe2+ elevated levels of oxidative stress markers and components of ferroptosis in the retina, and killed PRs. Ferrostatin-1 failed to protect the retina from Fe2+-induced oxidative damage. To investigate the mechanism of Fe2+-induced RPE autofluorescence, rd10 mutant mice aged 6 weeks, with almost total loss of PRs, were given intravitreal Fe2+ or Fe3+ injections: neither induced RPE autofluorescence. This result suggests Fe2+-induced RPE autofluorescence in wild-type mice resulted from phagocytosed, oxidized outer segments. Together these data suggest that intraretinal Fe2+ causes PR oxidative stress, leading to PR death and RPE autofluorescence.

DeshadowGAN: A Deep Learning Approach to Remove Shadows from Optical Coherence Tomography Images

PurposeTo remove blood vessel shadows from optical coherence tomography (OCT) images of the optic nerve head (ONH).MethodsVolume scans consisting of 97 horizontal B-scans were acquired through the center of the ONH using a commercial OCT device for both eyes of 13 subjects. A custom generative adversarial network (named DeshadowGAN) was designed and trained with 2328 B-scans in order to remove blood vessel shadows in unseen B-scans. Image quality was assessed qualitatively (for artifacts) and quantitatively using the intralayer contrast—a measure of shadow visibility ranging from 0 (shadow-free) to 1 (strong shadow). This was computed in the retinal nerve fiber layer (RNFL), the inner plexiform layer (IPL), the photoreceptor (PR) layer, and the retinal pigment epithelium (RPE) layer. The performance of DeshadowGAN was also compared with that of compensation, the standard for shadow removal.ResultsDeshadowGAN decreased the intralayer contrast in all tissue layers. On average, the intralayer contrast decreased by 33.7 ± 6.81%, 28.8 ± 10.4%, 35.9 ± 13.0%, and 43.0 ± 19.5% for the RNFL, IPL, PR layer, and RPE layer, respectively, indicating successful shadow removal across all depths. Output images were also free from artifacts commonly observed with compensation.ConclusionsDeshadowGAN significantly corrected blood vessel shadows in OCT images of the ONH. Our algorithm may be considered as a preprocessing step to improve the performance of a wide range of algorithms including those currently being used for OCT segmentation, denoising, and classification.Translational RelevanceDeshadowGAN could be integrated to existing OCT devices to improve the diagnosis and prognosis of ocular pathologies.

Novel AAV44.9-Based Vectors Display Exceptional Characteristics for Retinal Gene Therapy.

The majority of inherited retinal diseases (IRDs) are caused by mutations in genes expressed in photoreceptors (PRs). The ideal vector to address these conditions is one that transduces PRs in large areas of retina with the smallest volume/lowest titer possible, and efficiently transduces foveal cones, the cells responsible for acute, daylight vision that are often the only remaining area of functional retina in IRDs. The purpose of our study was to evaluate the retinal tropism and potency of a novel capsid, AAV44.9, and rationally designed derivatives thereof. We found that AAV44.9 and AAV44.9(E531D) transduced retinas of subretinally injected (SRI) mice with higher efficiency than did benchmark AAV5- and AAV8-based vectors. In macaques, highly efficient cone and rod transduction was observed following submacular and peripheral SRI. AAV44.9- and AAV44.9(E531D)-mediated GFP fluorescence extended laterally well beyond SRI bleb margins. Notably, extrafoveal injection (i.e., fovea not detached during surgery) led to transduction of up to 98% of foveal cones. AAV44.9(E531D) efficiently transduced parafoveal and perifoveal cones, whereas AAV44.9 did not. AAV44.9(E531D) was also capable of restoring retinal function to a mouse model of IRD. These novel capsids will be useful for addressing IRDs that would benefit from an expansive treatment area.

Mouse Models of Inherited Retinal Degeneration with Photoreceptor Cell Loss.

Inherited retinal degeneration (RD) leads to the impairment or loss of vision in millions of individuals worldwide, most frequently due to the loss of photoreceptor (PR) cells. Animal models, particularly the laboratory mouse, have been used to understand the pathogenic mechanisms that underlie PR cell loss and to explore therapies that may prevent, delay, or reverse RD. Here, we reviewed entries in the Mouse Genome Informatics and PubMed databases to compile a comprehensive list of monogenic mouse models in which PR cell loss is demonstrated. The progression of PR cell loss with postnatal age was documented in mutant alleles of genes grouped by biological function. As anticipated, a wide range in the onset and rate of cell loss was observed among the reported models. The analysis underscored relationships between RD genes and ciliary function, transcription-coupled DNA damage repair, and cellular chloride homeostasis. Comparing the mouse gene list to human RD genes identified in the RetNet database revealed that mouse models are available for 40% of the known human diseases, suggesting opportunities for future research. This work may provide insight into the molecular players and pathways through which PR degenerative disease occurs and may be useful for planning translational studies.

Differential day-night expression of tight junction components in murine retinal pigment epithelium

Strong communication and interaction between the retinal pigment epithelium (RPE) and the photoreceptor (PR) cells is essential for vision. RPE cells are essential for supporting and maintaining PR cells by transporting nutrients, waste products and ions, and phagocytosing photoreceptor outer segments (POS). POS phagocytosis follows a circadian pattern, taking place in the morning in human, mice and other organisms. However, it remains unknown whether other RPE processes follow a daily rhythm. To study the daily rhythm of RPE cells, we isolated murine RPE cells at six different time points during a 24 h period, after which RNA was isolated and sequenced. Murine RPE flatmounts were isolated at four different time points to study daily rhythm in protein abundance and localisation. EnrichR pathway analysis resulted in 13 significantly-enriched KEGG pathways (p < 0.01) of which seven showed a large number of overlapping genes. Several genes were involved in intracellular trafficking, possibly playing a role in nutrient transport, POS phagocytosis or membrane protein trafficking, with different expression patterns during the day-night cycle. Other genes were involved in actin cytoskeleton building, remodelling and crosslinking and showed a high expression in the morning, suggesting actin cytoskeleton remodelling at this time point. Finally, tight junction proteins Cldn2 and Cldn4 showed a difference in RNA and protein expression and tight junction localisation over time. Our study suggests that several important processes in the RPE follow a day-night rhythm, including intracellular trafficking, and processes involving the actin cytoskeleton and tight junctions. The differential protein localisation of Cldn2 in the RPE during the day-night cycle suggest that Cldn2 may facilitate paracellular water and sodium transport during the day.

Generation of a Retina Reporter hiPSC Line to Label Progenitor, Ganglion, and Photoreceptor Cell Types.

PurposeEarly in mammalian eye development, VSX2, BRN3b, and RCVRN expression marks neural retinal progenitors (NRPs), retinal ganglion cells (RGCs), and photoreceptors (PRs), respectively. The ability to create retinal organoids from human induced pluripotent stem cells (hiPSC) holds great potential for modeling both human retinal development and retinal disease. However, no methods allowing the simultaneous, real-time monitoring of multiple specific retinal cell types during development currently exist.MethodsCRISPR/Cas9-mediated homology-directed repair (HDR) in hiPSCs facilitated the replacement of the VSX2 (Progenitor), BRN3b (Ganglion), and RCVRN (Photoreceptor) stop codons with sequences encoding a viral P2A peptide fused to Cerulean, green fluorescent protein, and mCherry reporter genes, respectively, to generate a triple transgenic reporter hiPSC line called PGP1. This was accomplished by co-electroporating HDR templates and sgRNA/Cas9 vectors into hiPSCs followed by antibiotic selection. Functional validation of the PGP1 hiPSC line included the ability to generate retinal organoids, with all major retinal cell types, displaying the expression of the three fluorescent reporters consistent with the onset of target gene expression. Disaggregated organoids were also analyzed by fluorescence-activated cell sorting and fluorescent populations were tested for the expression of the targeted gene.ResultsRetinal organoids formed from the PGP1 line expressed appropriate fluorescent proteins consistent with the differentiation of NRPs, RGCs, and PRs. Organoids produced from the PGP1 line expressed transcripts consistent with the development of all major retinal cell types.Conclusions and Translational RelevanceThe PGP1 line offers a powerful new tool to study retinal development, retinal reprogramming, and therapeutic drug screening.

Light sensing in plant- and rock-associated black fungi

Fungi that share light-flooded habitats with phototrophs may profit from their excess photosynthetic products. But to cope with sunlight-associated stresses [e.g. high temperatures, UV radiation with associated DNA damage, accumulation of reactive oxygen species (ROS), desiccation and osmotic stresses] it is important for fungi to accurately sense and respond to changes in light. To test the hypothesis that light is an environmental cue that Ascomycota use to coordinate growth, stress responses as well as to establish pathogenic or symbiotic relationships, the photoreceptor (PR) distribution in species from different ecological niches was analysed. The genomes of black [dihydroxynaphthalene (DHN) melanin-containing] fungi from phyllosphere and exposed solid surfaces contain multiple photoreceptors (PRs). The plant pathogen Botrytis cinerea (Leotiomycetes) has a highly sophisticated photosensory and signalling system that helps to avoid light and to locate susceptible hosts. Rock-inhabiting Dothideomycetes and Eurotiomycetes including Knufia petricola possess equal numbers of PRs along with the same set of protective pigments. This similarity between black fungi from plant and rock surfaces suggests that photoperception and -regulation are important for fungi that receive nutrients through cooperation with phototrophs. Genetic tools for manipulating K. petricola exist and will be used to test this idea.

Morphological and functional indicators of retinal pigment epithelium and photoreceptor apparatus in inherited retinal diseases

To evaluate the relationship between the morphological and functional parameters of retinal pigment epithelium (RPE) and photoreceptors (PR) in inherited retinal diseases (IRD). The study included 52 patients (104 eyes), 23 of them with Stargardt Disease (STGD), 19 with cone-rod dystrophy (CRD), 10 with retinitis pigmentosa/pigmentary abiotrophy (RP) of comparable disease durations. All patients underwent standard and additional ophthalmological examination: fundus autofluorescence (AF), spectral optical coherence tomography (OCT), computer perimetry (CP), electro-oculography (EOG), Ganzfeld electroretinography (gERG). Comparison of the groups of IRD patients and groups according to the degree of RPE damage with the control group revealed an increase in differences in the EOG and gERG indicators as the area and depth of damage to the RPE and PR progressed. The patterns of changes in RPE and PR, the frequency of their occurrence with IRD in this patient sample are described. A moderate correlation was found between the amount of RPE loss and EOG light rise, as well as between the defect of the ellipsoid zone and the amplitude of α- and β-waves, the latency of β-wave of the gERG. Some patients showed a mismatch between a small defect of the ellipsoid zone and RPE with significant damage to the visual field and reduction of the EOG and gERG indicators. The obtained electrophysiological indicators revealed pathological changes in RPE and PR, more significant and widespread in some cases than it was shown with visualization methods. Weak and moderate correlations between visual acuity, and RPE damage and light sensitivity index with loss of ellipsoid zone were calculated. Modern methods of retinal examination can help obtain complete and versatile picture of morphological and functional state of the retina in IDR that supplement each other. EOG and gERG have capability to determine the degree of RPE and PR functions impairment including those cases when morphological studies are not sufficiently informative.

Traumatic “TERSON SYNDROME PLUS”: Pneumocephalocele with optic atrophy

Introduction. Terson Syndrome is subarachnoid hemorrhage (SAH) with sub retinal hemorrhage flowing through channel. Reduced vision in such fresh case is due to hemorrhage itself, blocking macula/other photo receptors in the long run macular cellophane retinopathy which causes profound visual loss. SAH causes neurological problems which can become a risk factor for evacuating blood from vitreous. Hypertension is commonest cause to cause Terson Syndrome, but trauma is also devastating cause as it can lead to irreversible visual consequences like total loss of perception of light or blindness. Aim. Here we describe a case of Terson Syndrome plus disease features SAH in frontal lobe. Description of the case. When there is traumatic pneumocephalocele, it gives space to blood to imbibe towards bony optic canal and form hematoma around nerve sheath which causes compression around the same and leads to optic atrophy. Optic nerve can be injured by direct traumatic dissection during road traffic accidents (RTA), but even without that blood may accumulate around optic nerve and in turn leads to formation of hematoma and subsequently pressure induced optic atrophy. Moreover, blood can slowly travel to sub hyaloid space/sub retinal space (beneath internal limiting membrane or sub ILM) with probable gliosis covering typical boat shaped blood as seen in this case. This sub ILM hemorrhage or gliosis may have resolved through three injections of Triamcinolone in the orbital floor (OFTA) near apex, but optic atrophy snatches vision. This protocol was followed to treat traumatic compressive (peri optic hematoma) optic neuropathy and traumatic retinopathy associated with sub hyaloid hemorrhage. Conclusion. Diagnosis of Terson syndrome plus disease was established by addressing all features on computed tomography (CT) scan and magnetic resonance imaging (MRI). Plus, features include pneumocephalus, optic nerve sheath hematoma, optic atrophy and gliosis over sub-hyaloid hemorrhage, typical boat shaped. The part of hemorrhage still endured as seen on optical coherence topography, but vision was lost by virtue of optic atrophy. OCT shows clot in sub hyaloid space.

Inner retinal preservation in the photoinducible I307N rhodopsin mutant mouse, a model of autosomal dominant retinitis pigmentosa.

Rod‐cone degenerations, for example, retinitis pigmentosa are leading causes of blindness worldwide. Despite slow disease progression in humans, vision loss is inevitable; therefore, development of vision restoration strategies is crucial. Among others, promising approaches include optogenetics and prosthetic implants, which aim to bypass lost photoreceptors (PRs). Naturally, the efficacy of these therapeutic strategies will depend on inner retinal structural and functional preservation. The present study shows that in photoinducible I307N rhodopsin mice (Translational Vision Research Model 4 [Tvrm4]), a 12k lux light exposure eliminates PRs in the central retina in 1 week, but interneurons and their synapses are maintained for as long as 9 weeks postinduction. Despite bipolar cell dendritic retraction and moderate loss of horizontal cells, the survival rate of various cell types is very high. Significant preservation of conventional synapses and gap junctions in the inner plexiform layer is also observed. We found the number of synaptic ribbons to gradually decline and their ultrastructure to become transiently abnormal, although based on our findings intrinsic retinal architecture is maintained despite complete loss of PRs. Unlike common rodent models of PR degeneration, where the disease phenotype often interferes with retinal development, in Tvrm4 mice, the degenerative process can be induced after retinal development is complete. This time course more closely mimics the timing of disease onset in affected patients. Stability of the inner retina found in these mutants 2 months after PR degeneration suggests moderate, stereotyped remodeling in the early stages of the human disease and represents a promising finding for prompt approaches of vision restoration.

Non‐invasive, needle‐free drug delivery for treatment of retinal degeneration on Bardet‐Biedl syndrome

AbstractPurposeTo develop a non‐invasive drug delivery system based on superparamagnetic nanoparticles (NP) for the treatment of retinal degeneration on Bardet‐Biedl syndrome (BBS). The delivery system will be used to deliver two small molecules, valproic acid (VPA) and Guanabenz (GBZ).MethodsA mixture of two NP (one loaded with VPA and other with GBZ) was applied as eyedrops on 14‐days old mice with the use of a magnetic field. The treatment was assessed in both wild type and a mice model for BBS (Bbs12−/−). The effects of the treatment were evaluated 2 weeks post treatment at 1‐month‐old. Retinal structure and function were analysed using electroretinogram recordings (ERG), histology and transmission electron microscopy (TEM).ResultsIn vitro studies showed no important toxic effects in cell cultures for retinal pigmented epithelium (RPE1) and 661W (murine photoreceptor precursor) after treatment with unloaded NP. Scotopic and photopic ERG recordings showed no toxic effect of the NP on photoreceptor (PR) function. TEM analysis indicated the presence of the NP in the PR layer of the retina after eyedrop application. TEM images showed a decrease in Endoplasmic Reticulum (ER) dilatation. The treated group with loaded NP showed an increase in the thickness of the inner nuclear layer, outer nuclear layer, inner PR segment and outer PR segment.ConclusionsThese results highlight the potential for these NP for non‐invasive drug delivery to the retina. Both the presence of the NP and biological effect were shown in our mouse model. The VPA and GBZ combination has already shown decrease of cell death mediated by ER stress in the Bbs12−/− model. This is an unmet clinical need with no curative treatment for retinal ciliopathies. The possibility of delaying PR death would increase the therapeutic window of gene therapy potentially increasing its effectivity.

Pharmacological modulation of the VCP/ERAD/proteasome axis rescues photoreceptor degeneration in P23H rhodopsin

AbstractPurposeOne of the most prevalent mutations in rhodopsin (Rh) causing autosomal dominant retinitis pigmentosa (adRP) RhP23H, results in its misfolding and its retention in the Endoplasmic Reticulum (ER). VCP is an ATP driven molecular checkpoint at the ER before these proteins are released from the ER towards plasma membrane. Misfolded proteins are selectively identified, retained at the ER and cleared by a process called ER‐associated degradation (ERAD). ERAD overload due to RhP23H misfolding causes cell death. As several pathways regulating protein homeostasis are associated with ERAD, we tested their specific impact on photoreceptor (PR) degeneration through pharmacological inhibition.MethodsOrganotypic cultures from RhP23H retinas of transgenic rats at PN9 cultured for 6 days and treated with different concentrations of pathway specific inhibitors: NMS‐873 interferes with quality control mechanisms by VCP inhibition; the proteasome inhibitor Bortezomib (BO) suppresses proteasomal degradation; the mannosidase inhibitor Kifunensine (KIF) blocks processing of glycoproteins, and Geldanamycin (GA) inhibits the protein‐folding mechanism in the ER. PR cell survival was assessed by counting cell rows and measuring the percentage of TUNEL‐positive cells. Rh distribution was checked using a specific antibody.ResultsVCP inhibitor caused strong and dose‐dependent protection at 5μM. Proteasome inhibition by BO significantly reduced the number of dying cells at 10μM. The number of PR cell rows in the ONL was increased by both. Only VCP inhibitor corrected aberrant inner segment distribution of Rh shifting it to physiological outer segment (OS). In contrary, KIF and GA accelerated PR cell degeneration.ConclusionsModulation of the VCP/ERAD/proteasome axis by inhibiting VCP and proteasome rescues PR cells in RhP23H. Inhibition of VCP activity establishes proper transport of Rh to the OS. These results strongly suggest that the VCP/ERAD/proteasome axis acts as a critical checkpoint in PR homeostasis.

Retinal neurodegeneration in systemic lupus erythematosus: layer by layer retinal study using optical coherence tomography

AbstractPurposeTo find early signs of retinal neurodegeneration by comparing the thickness of peripapillary retinal nerve fiber layer (pRNFL) and of all macular layers between systemic lupus erythematosus (SLE) patients without ophthalmologic manifestations and healthy controls. The effect of disease duration and systemic comorbidities was also studied.MethodsCross‐sectional study, in which retinal segmentation analysis with spectral domain‐optical coherence tomography (SD‐OCT) was performed. For pRNFL thickness, the global and six peripapillary sectors were determined. Each retinal layer thickness was calculated in the nine early treatment diabetic retinopathy study (ETDRS) subfields. Multiple regression analysis was performed.ResultsSixty‐eight eyes of 68 SLE patients and 50 eyes of 50 healthy controls were considered. pRNFL was significantly thinner in the SLE group globally (p = 0.026), in temporal superior (p = 0.007) and temporal (p = 0.037) sectors. Multivariable analysis in the SLE group revealed that chronic medication with antihypertensives, statins and anticoagulants were associated with a thinner pRNFL in some sectors (p &lt; 0.05). SLE patients presented a significant thinning in the photoreceptor (PR) layer in five ETDRS areas (p &lt; 0.05). Shorter disease duration was associated with greater photoreceptor thinning in all ETDRS subfields. Neuropsychiatric SLE, higher disease activity index and cardiovascular risk factors were associated with a thinner PR layer. No differences were observed in the remaining macular layers.ConclusionThis was the first study to perform OCT segmentation analysis of all retinal layers in SLE patients. These patients present a significant reduction in pRNFL and macular PR layer thickness. These differences were more pronounced in patients with poor systemic disease control, neuropsychiatric SLE and higher cardiovascular risk burden. Thus, there is evidence for the presence of early signs of retinal neurodegeneration in SLE.

Membrane-type frizzled-related protein regulates lipidome and transcription for photoreceptor function.

Molecular decision‐makers of photoreceptor (PRC) membrane organization and gene regulation are critical to understanding sight and retinal degenerations that lead to blindness. Using Mfrprd6mice, which develop PRC degeneration, we uncovered that membrane‐type frizzled‐related protein (MFRP) participates in docosahexaenoic acid (DHA, 22:6) enrichment in a manner similar to adiponectin receptor 1 (AdipoR1). Untargeted imaging mass spectrometry demonstrates cell‐specific reduction of phospholipids containing 22:6 and very long‐chain polyunsaturated fatty acids (VLC‐PUFAs) in Adipor1−/−and Mfrprd6 retinas. Gene expression of pro‐inflammatory signaling pathways is increased and gene‐encoding proteins for PRC function decrease in both mutants. Thus, we propose that both proteins are necessary for retinal lipidome membrane organization, visual function, and to the understanding of the early pathology of retinal degenerative diseases.

Are There Different Phenotypes in Geographic Atrophy of AMD? - Pilot Study on Differentiation Using Multimodal Imaging

Geographic atrophy (GA) in patients with age-related macular degeneration (AMD) involves a loss of photoreceptors (PR), retinal pigment epithelium (RPE) and choriocapillaris (CC). For treatment decisions, it is crucial to discern which of these layers the damage originates, subsequently spreading to the others. It has long been thought that the RPE, with its accumulation of lipofuscin, is the site of primary damage in the development of GA. However, histological studies have shown that in some patients, the PR are affected first, followed by secondary damage to the RPE and CC, and in others regression of the CC is the first manifestation. The aim of this study was to use multimodal imaging to determine the extent of the damage at the levels of the PR, RPE and CC, to characterise the individual phenotypic variations of GA and to investigate the corresponding functional impairment. Twenty eyes of 20 patients (mean age 78 years; 14 female, 6 male) with the clinical diagnosis of GA were examined by means of fundus autofluorescence (FAF) to evaluate the damage to the RPE, en face SD-OCT at the level of the PR to characterise the area of cell loss in this layer and OCT angiography (OCT-A, AngioVue, Optovue; 50 µm CC-segmentation with localization below the RPE) to assess regression of the CC. The affected area of each layer was measured. Best-corrected visual acuity (BCVA) test and fundus correlated automated 10° microperimetry (MAIA Microperimetry, CENTERVUE; 4-2 strategy, 68 stimuli) were performed in all patients. The results of these examinations were evaluated and correlated. All eyes showed a different extent of the areas of atrophy in the PR, RPE and CC. The layer with the largest area of atrophy was the RPE in 13 eyes (65%), the PR in 3 eyes (15%) and the CC in 4 eyes (20%). While the visual loss depended entirely on the presence of foveal sparing, microperimetry revealed a correlation between the extent of detectable functional deficit and the largest atrophic area. Multimodal imaging with FAF, en face OCT, OCT-A and a correlation with microperimetry enables a clinical phenotypic differentiation in GA as well as a more precise characterisation of the associated functional impairment. This confirms clinically the histologically demonstrated diversity of the damaged structure (PR, RPE or CC) in patients with GA. However, the variations identified in this pilot study must be confirmed in Reading Center-based larger cohorts. The approach described here may lead to differentiated consideration of the anatomical and functional aspects of the disease and turn out to be helpful in patient selection as well as in identifying and monitoring future therapeutic approaches.

Interactions between Dpr11 and DIP-γ control selection of amacrine neurons in Drosophila color vision circuits.

Drosophila R7 UV photoreceptors (PRs) are divided into yellow (y) and pale (p) subtypes. yR7 PRs express the Dpr11 cell surface protein and are presynaptic to Dm8 amacrine neurons (yDm8) that express Dpr11's binding partner DIP-γ, while pR7 PRs synapse onto DIP-γ-negative pDm8. Dpr11 and DIP-γ expression patterns define 'yellow' and 'pale' color vision circuits. We examined Dm8 neurons in these circuits by electron microscopic reconstruction and expansion microscopy. DIP-γ and dpr11 mutations affect the morphologies of yDm8 distal ('home column') dendrites. yDm8 neurons are generated in excess during development and compete for presynaptic yR7 PRs, and interactions between Dpr11 and DIP-γ are required for yDm8 survival. These interactions also allow yDm8 neurons to select yR7 PRs as their appropriate home column partners. yDm8 and pDm8 neurons do not normally compete for survival signals or R7 partners, but can be forced to do so by manipulation of R7 subtype fate.

Ganglion cell layer thickening in well-controlled patients with type 1 diabetes: an early sign for diabetic retinopathy?

PurposeTo evaluate early changes in retinal layers using optical coherence tomography (OCT) in patients with long‐standing type 1 diabetes (DM1) receiving intensified insulin therapy.MethodsIn a cross‐sectional case–control study 150 patients with DM1 and 150 age‐ and sex‐matched healthy control participants underwent OCT imaging. Scans of both eyes were analysed for different layers (NFL, GCL (+IPL), INL, outer layer complex (OLC, including OPL, ONL and ELM) and photoreceptors (PR)) in all subfields of an ETDRS grid. All analyses were performed semi‐automatically using custom software by certified graders of the Vienna Reading Center. ANOVA models were used to compare the mean thickness of the layers between patients and controls.ResultsSix hundred eyes with 512 datapoints in 49 b‐scans in each OCT were analysed. Mean thickness in patients/controls was 31.35 μm/30.65 μm (NFL, p = 0.0347), 76.7 μm/73.15 μm (GCL, p ≤ 0.0001), 36.29 μm/37.13 μm (INL, p = 0.0116), 114.34 μm/112.02 μm (OLC, p < 0.0001) and 44.71 μm/44.69 μm (PR, p = 0.9401). When evaluating the ETDRS subfields separately for clinically meaningful hypotheses, a significant swelling of the GCL in patients could be found uniformly and a central swelling for the OLC, whereas the distribution of NFL and INL thickening suggests that their statistical significance was not clinically relevant.ConclusionThese preliminary results demonstrate that preclinical retinal changes in patients with long‐standing DM1 can be found by retinal layer evaluation. However, the changes are layer‐specific, with significant thickening of the GCL and less so of the OLC suggesting a role as an early sign for diffuse swelling and the evolution of DME even in well‐controlled diabetes.