Retinal Structure Research Articles

ODDF-Net: Multi-object segmentation in 3D retinal OCTA using optical density and disease features

Automatic extraction of retinal structures, including Retinal Capillaries (RC), Retinal Arteries (RA), Retinal Veins (RV), and the Foveal Avascular Zone (FAZ), is crucial for the diagnosis and treatment of ocular diseases. This paper presents ODDF-Net, a segmentation network leveraging optical density and disease features, for the simultaneous 2D segmentation of RC, RA, RV, and FAZ in 3D Optical Coherence Tomography Angiography (OCTA). We introduce the concept of optical density to generate additional input images, enhancing the specificity for distinguishing arteries and veins. Our network employs a decoupled segmentation head to separate independent features of each object from shared features by focusing on object boundaries. Given the impact of ocular diseases on the morphology of retinal objects, we designed an auxiliary classification head and a cross-dimensional feature fusion module to model the relationship between various diseases and changes in retinal structures. Extensive experiments on two subsets of the OCTA-500 dataset demonstrate that ODDF-Net outperforms state-of-the-art methods, achieving mean intersection over union ratios of 88.17% and 82.80%. The source code is available at https://github.com/y8421036/ODDF-Net.

In vivo photoreceptor base editing ameliorates rhodopsin-E150K autosomal-recessive retinitis pigmentosa in mice

Rhodopsin, the prototypical class-A G-protein coupled receptor, is a highly sensitive receptor for light that enables phototransduction in rod photoreceptors. Rhodopsin plays not only a sensory role but also a structural role as a major component of the rod outer segment disc, comprising over 90% of the protein content of the disc membrane. Mutations in RHO which lead to structural or functional abnormalities, including the autosomal recessive E150K mutation, result in rod dysfunction and death. Therefore, correction of deleterious rhodopsin mutations could rescue inherited retinal degeneration, as demonstrated for other visual genes such as RPE65 and PDE6B. In this study, we describe a CRISPR/Cas9 adenine base editing strategy to correct the E150K mutation and demonstrate precise in vivo editing in a Rho -E150K mouse model of autosomal recessive retinitis pigmentosa (RP). Using ultraviolet-visible spectroscopy, mass spectrometry, and the G-protein activation assay, we characterized wild-type rhodopsin and rhodopsin variants containing bystander base edits. Subretinal injection of dual-adeno-associated viruses delivering our base editing strategy yielded up to 44% Rho correction in homozygous Rho -E150K mice. Injection at postnatal day 15, but not later time points, restored rhodopsin expression, partially rescued retinal function, and partially preserved retinal structure. These findings demonstrate that in vivo base editing can restore the function of mutated structural and functional proteins in animal models of disease, including rhodopsin-associated RP and suggest that the timing of gene-editing is a crucial determinant of successful treatment outcomes for degenerative genetic diseases.

Prolonged Photobiomodulation with Deep Red Light Mitigates Incipient Retinal Deterioration in a Mouse Model of Type 2 Diabetes

Diabetic retinopathy is a prevalent complication of type 2 diabetes mellitus, characterized by progressive damage to the retinal structure and function. Photobiomodulation therapy, using red or near-infrared light, has been proposed as a non-invasive intervention to mitigate retinal damage, but has been tested generally with short-term stimuli. This study aimed to evaluate the effects of prolonged photobiomodulation with deep red light on retinal structure and function in a type 2 diabetes mouse model. Transgenic LepRdb/J (db/db) mice were exposed to photobiomodulation therapy via LED devices emitting 654 nm light for 12 h daily over ten weeks and compared to untreated mice. Retinal function was evaluated by flash electroretinography, while structural changes were assessed through histology and immunohistochemistry to detect astro- and microgliosis. At 33 weeks of age, db/db mice were obese and severely diabetic, but exhibited only incipient indicators of retinal deterioration. Electroretinogram b-wave peak latencies were prolonged at intermediate flash intensities, while the outer plexiform layer displayed significantly elevated IBA1 expression. Photobiomodulation therapy prevented these two markers of early retinal deterioration but had no effect on other morphological and functional parameters. Photobiomodulation is well-tolerated and maintains potential as a complementary treatment option for diabetic retinopathy but requires further optimization of therapeutic settings and combinatory treatment approaches.

Multiscale attention-over-attention network for retinal disease recognition in OCT radiology images

Retinal disease recognition using Optical Coherence Tomography (OCT) images plays a pivotal role in the early diagnosis and treatment of conditions. However, the previous attempts relied on extracting single-scale features often refined by stacked layered attentions. This paper presents a novel deep learning-based Multiscale Feature Enhancement via a Dual Attention Network specifically designed for retinal disease recognition in OCT images. Our approach leverages the EfficientNetB7 backbone to extract multiscale features from OCT images, ensuring a comprehensive representation of global and local retinal structures. To further refine feature extraction, we propose a Pyramidal Attention mechanism that integrates Multi-Head Self-Attention (MHSA) with Dense Atrous Spatial Pyramid Pooling (DASPP), effectively capturing long-range dependencies and contextual information at multiple scales. Additionally, Efficient Channel Attention (ECA) and Spatial Refinement modules are introduced to enhance channel-wise and spatial feature representations, enabling precise localization of retinal abnormalities. A comprehensive ablation study confirms the progressive impact of integrated blocks and attention mechanisms that enhance overall performance. Our findings underscore the potential of advanced attention mechanisms and multiscale processing, highlighting the effectiveness of the network. Extensive experiments on two benchmark datasets demonstrate the superiority of the proposed network over existing state-of-the-art methods.

Enhancing Late Retinopathy of Prematurity Outcomes with Fresh Bone Marrow Mononuclear Cells and Melatonin Combination Therapy.

Retinopathy of prematurity (ROP) is a vasoproliferative disease affecting premature neonates with life-lasting impacts. This study aims to investigate the long-term functional outcomes and alterations in neural retina architecture following the intravitreal transplantation of bone marrow mononuclear cells (BMMNC) in the rat models of ROP, and to evaluate the effect of adjunctive therapy with melatonin. 32 neonate rats were employed. The ROP model was developed in 10 neonatal rats, and two were assigned as control. The ROP models received BMMNC suspension, containing 1.2 × 105cells, in their right eye, and normal saline in left at p12. Five ROP rats received 12.5 mg/kg melatonin orally for five days (p12 to p17). Optical coherence tomography (OCT) and electroretinography (ERG) were performed on p47. Eyes were then harvested on p47, and after six months for histology, immunofluorescence (anti-calbindin, anti-PKC, and anti-Brn3), and immunohistochemistry (synaptophysin). Cell therapy alone and with melatonin increased retinal thickness, and improved oscillatory potentials on ERG. Combination therapy increased horizontal and retinal ganglion cell populations. All treatments improved synaptic maturity in the inner plexiform layer, but only combination therapy was effective on the outer plexiform layer. Melatonin and BMMNCs combination therapyeffectively ameliorates retinal structural and functional deficits at later ROP stages, without causing severe adverse effects.It significantly increases the survival of post-receptor retinal neurons and preserves retinal synaptic structures in the long term, highlighting the promising potential of this novel combination therapy approach to minimize visual deficits in ROP patients.

112° field of view high-resolution swept-source OCT angiography for rat retinas.

This study introduces an ultra-wide field (UWF) and high-resolution swept-source optical coherence tomographic angiography (OCTA) system for rat retinal imaging. Using an asymmetrical optics design, the system achieves unprecedented details of retinal structures and vascular plexuses over a large field of view (112°) in a single-shot acquisition. Views of single-nerve fiber bundles and single capillary vessels are consistently visible over a 112° field of view. The system has a long imaging range and high penetration and allows a full view of vitreous hyaloid vessels, retina, choroid, sclera, and posterior ciliary arteries, down to sub-sclera connective tissues. In a longitudinal study of oxygen-induced retinopathy (OIR) in rats, the system successfully revealed the progression and regression of OIR-related vascular pathologies, such as ischemia, neovascularization, and tortuosity. To our knowledge, this novel UWF-OCT/OCTA prototype designed for rat retinal imaging will be a vital tool for monitoring disease progression and evaluating therapeutic interventions in preclinical models.

A Method for Retina Segmentation by Means of U-Net Network

Retinal image segmentation plays a critical role in diagnosing and monitoring ophthalmic diseases such as diabetic retinopathy and age-related macular degeneration. We propose a deep learning-based approach utilizing the U-Net network for the accurate and efficient segmentation of retinal images. U-Net, a convolutional neural network widely used for its performance in medical image segmentation, is employed to segment key retinal structures, including the optic disc and blood vessels. We evaluate the proposed model on a publicly available retinal image dataset, demonstrating interesting performance in automatic retina segmentation, thus showing the effectiveness of the proposed method. Our proposal provides a promising method for automated retinal image analysis, aiding in early disease detection and personalized treatment planning.

Blue light reflectance imaging in non-perfusion areas detection: insights from multimodal analysis

DesignA retrospective, cross-sectional image analysis using a convenience sample.SubjectsFive cases selected based on the availability of comprehensive imaging data.MethodsThis study involved a retrospective review of images from five cases, focusing on the use of retinal monochromatic blue light reflectance (BLR) imaging to detect non-perfusion areas. Two cases of sickle-cell retinopathy demonstrated peripheral retinal non-perfusion identified through widefield fluorescein angiography. Three other cases—one with branch retinal vein occlusion, one with branch retinal artery occlusion, and one presenting paracentral acute middle maculopathy showed focal macular non-perfusion detected by structural OCT and OCTA. The areas of nonperfused retinal tissue, confirmed by fluorescein angiography, OCT, and OCTA, were then correlated with findings from the BLR image. This correlation aimed to identify any potential associations between these imaging modalities.Main outcome measuresEnhance understanding of the utilization of retinal monochromatic BLR images as a non-perfusion biomarker.ResultsThe perfusion defects identified through fluorescein angiography were qualitatively correlated with hypo-reflective regions observed in the BLR images. A notable correlation was also observed between the OCTA deep capillary plexus findings and the BLR images. Additionally, areas of retinal thinning identified on structural OCT thickness maps corresponded with the hypo-reflective regions in the BLR images. This indicates the potential of BLR in identifying non-perfused retinal areas.ConclusionsThis study reinforces the evidence, through OCT, OCTA, and angiographic correlation, that the BLR can effectively identify areas of retinal non-perfusion in a non-invasive manner. Further research is warranted to assess the method’s sensitivity, specificity, and limitations. While the interaction of blue light with the retina, leading to specular reflections and scattering, is established, this research represents a pioneering effort in suggesting which specific retinal structures may be implicated in this phenomenon. This novel insight opens avenues for deeper exploration into the underlying mechanisms and potential clinical applications of utilizing the BLR imaging technique for assessing retinal vascular abnormalities.

Evaluating the effects of three anesthetic regimens on retinal structure and function in the living mouse eye using a customized bimodal OCT/ORG system

Abstract Optical coherence tomography (OCT) is a non-invasive imaging technique that generates cross-sectional images of biological tissues at millimeter-level penetration depth with micron-level resolution. In this study, we developed a custom spectral-domain OCT system equipped with a flash stimulus module for dual-modality imaging of mouse retinal structure and function. The axial resolution of the system reached ∼2.7/2 μm in air/tissue following optimization and calibration. We deployed this system to conduct in vivo OCT structural imaging and optoretinography (ORG) functional assessment of mouse retina to investigate the effects of three anesthesia regimens. Statistical analysis of the results indicates that anesthesia results in a smaller relative intensity of inner/outer segment junction and external limiting membrane and a thickening of the outer segment compared to an awake state. Meanwhile, significant differences were observed in the effects on retinal structure and function between isoflurane and tribromoethanol anesthesia, while differences between isoflurane and esketamine+xylazine groups were small. These findings suggest that the type of anesthetic should be taken into account when comparing imaging results across studies reported in the literature.

Comparison of Vessel Density and Retinal Sensitivity After Scleral Buckling and Phacovitrectomy in the Management of Macula-on Primary Rhegmatogenous Retinal Detachment.

The choice of surgical method for rhegmatogenous retinal detachment (RRD) may have a significant impact on the retina. In this study, we aimed to compare retinal function and structure after scleral buckling (SB) and phacovitrectomy (phaco-PPV) for macula-on RRD. This cross-sectional study included patients who underwent anatomically successful repair of macula-on RRD managed with SB (n=35) and phaco-PPV (n=35) between 2019-2023. All participants were examined within 6-20 months of surgery to evaluate the retinal structure using spectral domain optical coherence tomography (SD-OCT) and vessel density (VD) by OCT angiography (OCTA). Best-corrected visual acuity (BCVA) and microperimetry (MP) tests were used to assess the retinal function. Analysis of the microvascular network with OCTA between eyes after surgery and healthy eyes showed a decrease in VD. Significant changes in the superficial vascular plexus (SVP) and deep vascular plexus (DVP) were observed only in eyes after SB surgery (p <0.001 and p=0.02, respectively). Analysis of retinal function assessed by MP showed a significant decrease (p<0.05) in retinal sensitivity after phaco-PPV (24.81±2.25 dB) and SB (24.18±2.14 dB) operations compared to the healthy control group (25.97 ± 1.51 dB), whereas postoperative BCVA showed no differences (p>0.05). Changes in retinal sensitivity were accompanied by impairment of the microvascular network in the eyes after SB and phaco-PPV surgeries due to macula-on RRD. Disorders were more pronounced in eyes following SB surgery, possibly secondary to mechanical stress.

A multimodal imaging approach to investigate retinal oxygen and vascular dynamics, and neural dysfunction in bietti crystalline dystrophy

BackgroundThis study aimed to explore retinal changes in Bietti crystalline dystrophy (BCD) patients, including retinal metabolism, blood flow, vascular remodeling, and pupillary light reflex (PLR) abnormalities. MethodsThis cross-sectional study included 120 eyes from BCD patients and 120 eyes from healthy controls, utilizing a multimodal imaging system (MEFIAS 3200, SYSEYE, Chongqing, China) to evaluate retinal oxygenation, blood flow, vascular structure, and PLR. Measurements included oxygen saturation, blood flow velocity, vessel diameters, and pulsatility metrics. PLR parameters were assessed under specific light stimuli. ResultsBCD patients demonstrated significantly higher retinal oxygen saturation and content, but lower oxygen utilization and metabolism compared to controls, with more pronounced declines in those over 40 years old. Vascular parameters revealed smaller external diameters and larger lumen diameters, indicating vascular remodeling. Retinal blood flow was lower, while the resistivity index was higher in BCD patients. Additionally, PLR abnormalities were noted, including reduced constriction amplitude, pupil constriction ratio, constriction duration, and maximum constriction velocity, along with prolonged latency were observed in BCD patients. ConclusionBCD patients had significant retinal and vascular changes, along with PLR impairments, especially in patients over 40. More targeted interventions should be focused in future research.

Multi-task OCTA image segmentation with innovative dimension compression

Optical Coherence Tomography Angiography (OCTA) plays a crucial role in the early detection and continuous monitoring of ocular diseases, which relies on accurate multi-tissue segmentation of retinal images. Existing OCTA segmentation methods typically focus on single-task designs that do not fully utilize the information of volume data in these images. To bridge this gap, our study introduces H2C-Net, a novel network architecture engineered for simultaneous and precise segmentation of various retinal structures, including capillaries, arteries, veins, and the fovea avascular zone (FAZ). At its core, H2C-Net consists of a plug-and-play Height-Channel Module (H2C) and an Enhanced U-shaped Network (GPC-Net). The H2C module cleverly converts the height information of the OCTA volume data into channel information through the Squeeze operation, realizes the lossless dimensionality reduction from 3D to 2D, and provides the "Soft layering" information by unidirectional pooling. Meanwhile, in order to guide the network to focus on channels for training, U-Net is enhanced with group normalization, channel attention mechanism, and Parametric Rectified Linear Unit (PReLU), which reduces the dependence on batch size and enhances the network's ability to extract salient features. Extensive experiments on two subsets of the publicly available OCTA-500 dataset have shown that H2C-Net outperforms existing state-of-the-art methods. It achieves average Intersection over Union (IoU) scores of 82.84 % and 88.48 %, marking improvements of 0.81 % and 1.59 %, respectively. Similarly, the average Dice scores are elevated to 90.40 % and 93.76 %, exceeding previous benchmarks by 0.42 % and 0.94 %. The proposed H2C-Net exhibits excellent performance in OCTA image segmentation, providing an efficient and accurate multi-task segmentation solution in ophthalmic diagnostics. The code is publicly available at: https://github.com/IAAI-SIT/H2C-Net.

Abnormal outer and inner retina in a mouse model of Huntington's disease with age.

Huntington's disease (HD) is a progressive neurodegenerative disorder characterized by motor dysfunction and cognitive decline. While retinal abnormalities have been documented in some HD patients and animal models, the nature of these abnormalities-specifically whether they originate in the inner or outer retina-remains unclear, particularly regarding their progression with age. This study investigates the retinal structure and function in HD transgenic mice (R6/1) compared to C57BL/6 J control mice at 2, 4, and 6 months of age, encompassing both pre-symptomatic and symptomatic stages of HD. Pathological assessments of the striatum and evaluations of motor function confirmed significant HD-related alterations in R6/1 mice at 6 months. Visual function was subsequently analyzed, accompanied by immunofluorescent staining of retinal and optic nerve tissues over time. Our findings revealed that R6/1 mice exhibited pronounced HD symptoms at 6 months, characterized by neuronal loss in the striatum and impaired locomotor abilities. Functionally, visual acuity declined at 6 months, while retinal light responses began to deteriorate by 4 months. Structurally, R6/1 mice demonstrated a global reduction in cone opsin expression as early as 2 months, with a decrease in rhodopsin levels at 4 months, alongside a thinner retinal structure compared to controls. Notably, rod bipolar cell populations were decreased at 6 months, exhibiting shorter dendritic branches and reduced synaptic connections with photoreceptors in the outer retina. Additionally, ganglion cell numbers in the inner retina decreased at 6 months, accompanied by aberrant neural fibers in the optic nerve. Microglial activation was evident at 4 months, while astrocytic activation was observed at 6 months. Aggregates of mutant huntingtin (mHTT) were first detected in the ganglion cell layer and optic nerve at 2 months, subsequently disseminating throughout all retinal layers with advancing age. These results indicate that retinal pathology in R6/1 mice manifests earlier in the outer retina than in the inner retina, which does not align with the progression of mHTT aggregation. Consequently, the R6/1 mouse retina may serve as a more effective model for elucidating the mechanisms underlying HD and evaluating potential therapeutic strategies, rather than functioning as an early diagnostic tool for the disease.

Ginsenoside Rg3 Improved Age-Related Macular Degeneration Through Inhibiting ROS-Mediated Mitochondrion-Dependent Apoptosis In Vivo and In Vitro.

Age-related macular degeneration (AMD) is marked by a progressive loss of central vision and is the third leading cause of irreversible blindness worldwide. The exact mechanisms driving the progression of this macular degenerative condition remain elusive, and as of now, there are no available preventative measures for dry AMD. According to ancient records, ginseng affects the eyes by brightening them and enhancing wisdom. Modern pharmacological research shows that the active ingredients in ginseng, ginsenosides, may be used to prevent or improve eye diseases that threaten vision. Some articles have reported that ginsenoside Rg3 can treat diabetic retinopathy in mice, but no reports exist on its effects and mechanisms in AMD. Therefore, the role and mechanism of ginsenoside Rg3 in AMD warrant further study. This study aims to investigate the effects of Rg3 on AMD and its underlying molecular mechanisms. We established a mouse model of AMD to examine the impact of ginsenoside Rg3 on NaIO3-induced apoptosis in the retina and to explore the related intrinsic mechanisms. The in vivo results indicated that ginsenoside Rg3 prevents NaIO3-induced apoptosis in retinal pigment epithelial cells by inhibiting reactive oxygen species production and preventing the reduction in mitochondrial membrane potential. Additionally, we assessed the levels of protein expression within the apoptosis pathway. Ginsenoside Rg3 decreased the expression of Bax, cleaved caspase-3, and cleaved caspase-9 proteins. Additionally, it increased the expression of Bcl-2 by decreasing P-JNK levels. Moreover, our in vivo results showed that ginsenoside Rg3 enhanced retinal structure, increased the relative thickness of the retina, and decreased the extent of disorganization in both the inner and outer nuclear layers. Ginsenoside Rg3 may safeguard the retina against NaIO3-induced cell apoptosis by attenuating reactive-oxygen-species-mediated mitochondrial dysfunction, in which the JNK signaling pathway is also involved. These findings suggest that ginsenoside Rg3 has the potential to prevent or attenuate the progression of AMD and other retinal pathologies associated with NaIO3-mediated apoptosis.

Evaluation of retinal nerve fiber layer and choroidal structure in obese children and adolescents

Aim: Obesity-related vascular damage and endothelial dysfunction have deleterious effects on the ocular vasculature. It was aimed to examine optical coherence tomography (OCT) parameters in obese and overweight children and to define their relationship with metabolic markers in this study. Methods: The patient group consisted of 26 obese, 24 overweight patients aged between 8 and 18 years. The control group consisted of 25 healthy children with normal body mass index (BMI). This was a cross-sectional observational study. Serum glucose, lipid parameters, and homeostasis model assessment of insulin resistance (HOMA-IR) were investigated. Measurement of choroidal thickness was performed with Cirrus HD-OCT (Carl Zeiss Meditec Inc., Dublin, CA, USA). Retinal nerve fiber layer (RNFL) thickness was determined by an automatic computer algorithm without the need for user measurement. Results: There were no differences in subfoveal, nasal, temporal choroidal thickness, and RNFL between obese, overweight, and control groups (p&gt;0.05). A positive (linear) moderate relationship was found between RNFL and the HOMA-IR of 26 patients in the obese group (r=0.389) (p=0.049). A positively weak correlation was found between height and RNFL in obese patient group (r=0.264, p=0.028). Conclusion: In the study, RNFL thickness increased as HOMA-IR level increased in obese children and adolescents. RNFL decreased as the height increased in obese children and adolescents. We believe that more comprehensive data about the effect of obesity on RNFL and choroidal thickness will be obtained with prospective studies in which the obese patient group with insulin resistance is taken separately and disease durations are defined, and long-term patient follow-up is performed.

Multi-Characteristic Opsin Therapy to Functionalize Retina, Attenuate Retinal Degeneration, and Restore Vision in Mouse Models of Retinitis Pigmentosa.

Retinal degeneration 1 and 10 (rd1 and rd10) mice are useful animal models of retinitis pigmentosa (RP) with rapidly and slowly progressive pathologies, respectively. Our study aims were to determine the effect of adeno-associated viral vector 2 (AAV2)-delivered multi-characteristic opsin (MCO-010; under the control of a metabotropic glutamate receptor-6 promoter enhancer) on the morphological and functional characteristics of vision in both rd1 and rd10 mice. Various retinal measures of MCO-010 transduction and electrophysiological, behavioral, and other routine blood analyses were performed in the rd1 and/or rd10 mice after intravitreal injection of 1µL of MCO-010 or AAV2 vehicle. Functional tests included electroretinogram, visually evoked potential, and behavior assay (optomotor and water maze). Retinal thickness, intraocular pressure, and plasma cytokine levels were also determined. Following intravitreal MCO-010 injection, approximately 80% of bipolar cells were transduced in the retina, and no alterations in retinal thickness were observed at 4months post-injection. However, retinal thickness significantly decreased in control mice. MCO-010 treatment increased head movements and induced faster navigation of mice to the platform in a water-maze test. The MCO-010 gene therapy helped preserve visually evoked electrical response in the retina and visual cortex. No ocular toxicity, immunotoxicity, or phototoxicity was observed in the MCO-010-treated mice, even under chronic intense light conditions. Intravitreal MCO-010 was well tolerated in rd1 and rd10 mice models of RP, and it appeared to attenuate retinal photoreceptor degeneration based on retinal structure and functional outcome measures. As reported here, optogenetic treatment of the inner retina attenuates further retinal degeneration in addition to photosensitizing higher order neurons, and this disease-modifying aspect should be evaluated in optogenetic clinical trials.

Splice Variant of Retinal G-Protein-Coupled Receptor Deletion-Mediated Dysregulation of Autophagy Increases the Susceptibility to Age-Related Macular Degeneration-Like Defects

Introduction: The splice variant of retinal G-protein-coupled receptor deletion (RGR-d) is a persistent component of drusen and may be involved in the pathogenesis of dry age-related macular degeneration (AMD). Increasing evidence has demonstrated the critical role of autophagy in AMD. In this study, we investigated whether RGR-d disrupts autophagy in early dry AMD in vivo and in vitro. Methods: Fundus imaging and fluoroscopy were performed on RGR-d mice created by multiplex gene editing. The retina microstructure was evaluated by performing hematoxylin and eosin (H&E) staining as well as transmission electron microscopy (TEM). Retinal function was assessed by full-field electroretinography (ERG). After lentivirus transfection and stimulation, the permeability, phagocytosis, and tight junctions of ARPE-19 cells were evaluated. Western blotting of ATG5, Beclin-1, LC3II/I, and P62 was performed to detect the changes in autophagy pathways. Results: Atrophy and patchy penetrating hyperfluorescent foci, consistent with early AMD-like defects, were observed in the fundus of 12-month-old RGR-d mice. H&E staining of retinal tissues indicated thinning of each layer of the retinal structure. H&E staining of retinal tissues indicated thinning of each layer of the retinal structure. TEM analysis showed some diffuse granular deposits. And the morphology of choroidal microvascular endothelial cells was degraded and distorted. The morphology of the photoreceptor outer segments showed structural damage, and Bruch’s membrane was thickened. ERG indicated that the photoreceptor of RGR-d mice were dysfunctional. Changes in autophagy-related protein expression were observed in the retinal pigment epithelium and retinal neurepithelium, and autophagy regulation was decreased. Palmitic acid (PA) stimulation caused permeability, phagocytosis, and tight junction dysfunction in cells overexpressing RGR-d. Beclin-1 and LC3II/I expression levels were significantly decreased and that of P62 was elevated in RGR-d cells after PA stimulation. Conclusion: RGR-d disrupts the autophagy pathway, causing the development of an early AMD-like pathophysiology.

IL-33 protects retinal structure and function via mTOR/S6 signaling pathway in optic nerve crush

This study demonstrated the functions and molecular mechanisms of the IL-33/ST2 axis in experimental optic neuropathy. C57BL/6J mice were used to establish an optic nerve crush (ONC) model. ONC mice were administered with IL-33 intraperitoneal injection, with PBS vehicle as control. Immunofluorescence, quantitative RT-PCR, and western blot techniques were utilized to assess the expression of the IL-33/ST2 axis. The electroretinography (ERG), optical coherence tomography (OCT), H&E, and luxol fast blue were used to assess the structural and functional changes. Western blot was employed to detect the activation of the mTOR/S6 pathway. The IL-33 expression level in the inner nuclear layer of the retina in ONC mice reached its peak on day 3, accompanied by a significant increase in IL-33 receptor ST2 expression. IL-33 treatment promoted the survival of retinal ganglion cells, restored the thickness of inner retina layer (IRL), alleviated the demyelination of the optic nerve, and recovered the decreased amplitude of b-wave in ONC mice. Furthermore, administration of IL-33 activated the mTOR/S6 signaling pathway in RGCs, which was significantly suppressed in the ONC condition. This study indicated that boosting the IL-33/ST2/mTOR/S6 pathway can protect against structural and functional damage to the retina and optic nerve induced by ONC. As a result, the IL-33/ST2 axis holds potential as a therapeutic option for treating various optic neuropathies.

Multimodal photoacoustic microscopy and optical coherence tomography ocular biomarker imaging in Alzheimer's disease in mice

Alzheimer's disease (AD) is a neurodegenerative disease characterized by amyloid beta (Aβ)-containing extracellular plaques and tau-containing intracellular neurofibrillary tangles. Reliable and more accessible biomarkers along with associated imaging methods are essential for early diagnosis and to develop effective therapeutic interventions. Described here is an integrated photoacoustic microscopy (PAM) and optical coherence tomography (OCT) dual-modality imaging system for multiple ocular biomarker imaging in an AD mouse model. Anti-Aβ-conjugated Au nanochains (AuNCs) were engineered and administered to the mice to provide molecular contrast of Aβ. The retinal vasculature structure and Aβ deposition in AD mice and wild-type (WT) mice were imaged simultaneously by dual-wavelength PAM. OCT distinguished significant differences in retinal layer thickness between AD and WT animals. With the unique ability of imaging the multiple ocular biomarkers via a coaxial multimodality imaging system, the proposed system provides a new tool for investigating the progression of AD in animal models, which could contribute to preclinical studies of AD.

Adaptive optics scanning laser ophthalmoscopy in a heterogenous cohort with Stargardt disease

Image based cell-specific biomarkers will play an important role in monitoring treatment outcomes of novel therapies in patients with Stargardt (STGD1) disease and may provide information on the exact mechanism of retinal degeneration. This study reports retinal image features from conventional clinical imaging and from corresponding high-resolution imaging with a confocal adaptive optics scanning laser ophthalmoscope (AOSLO) in a heterogenous cohort of patients with Stargardt (STGD1) disease. This is a prospective observational study in which 16 participants with clinically and molecularly confirmed STGD1, and 7 healthy controls underwent clinical assessment and confocal AOSLO imaging. Clinical assessment included short-wavelength and near-infrared fundus autofluorescence, spectral-domain optical coherence tomography, and macular microperimetry. AOSLO images were acquired over a range of retinal eccentricities (0°–20°) and mapped to areas of interest from the clinical images. A regular photoreceptor mosaic was identified in areas of normal or near normal retinal structure on clinical images. Where clinical imaging indicated areas of retinal degeneration, the photoreceptor mosaic was disorganised and lacked unambiguous cones. Discrete hyper-reflective foci were identified in 9 participants with STGD1 within areas of retinal degeneration. A continuous RPE cell mosaic at the fovea was identified in one participant with an optical gap phenotype. The clinical heterogeneity observed in STGD1 is reflected in the findings on confocal AOSLO imaging.