Photoreceptor Outer Segment Tips Research Articles

Polarization-resolved analysis of outer retinal bands: investigating ballistic and multiply scattered photons using full-field swept-source optical coherence tomography.

Precise interpretation of the anatomical origins of outer retinal optical coherence tomography (OCT) presents technical challenges owing to the delicate nature of the retina. To address this challenge, our study introduces a novel polarization-sensitive full-field swept-source OCT (FF-SS-OCT) that provides parallel-polarization and cross-polarization OCT measurements, predominantly capturing ballistically reflected photons and multiply scattered photons, respectively. Notably, parallel-polarization OCT unveils layer-like structures more effectively, including the inner plexiform layer (IPL) sub-layers, outer plexiform layer (OPL) sub-layers (in rod-dominant regions), and rod/cone outer segment (OS) tips, compared to cross-polarization OCT, where such sub-layers are not visible. Through a comparative analysis of parallel-polarization and cross-polarization OCT images of the outer retina, we discovered that the 2nd outer retinal OCT band results from contributions from both the ellipsoid zone (EZ) and the inner segment/outer segment (IS/OS) junction. Similarly, the 3rd outer retinal OCT band appears to reflect contributions from both the interdigitation zone (IZ) and photoreceptor OS tips. This polarization-sensitive approach advances our understanding of the origins of outer retinal OCT signals and proposes potential new biomarkers for assessing retinal health and diseases.

Senescence-induced alteration of circadian phagocytic activity of retinal pigment epithelium cell line ARPE-19

Renewal of retinal photoreceptor outer segments is conducted through daily shedding of distal photoreceptor outer segment tips and subsequent their phagocytosis by the adjacent retinal pigment epithelium (RPE) monolayer. Dysregulation of the diurnal clearance of photoreceptor outer segment tips has been implicated in age-related retinal degeneration, but it remains to be clarified how the circadian phagocytic activity of RPE cells is modulated by senescence. In this study, we used the human RPE cell line ARPE-19 to investigate whether hydrogen peroxide (H2O2)-induced senescence in ARPE-19 cells alters the circadian rhythm of their phagocytic activity. After synchronization of the cellular circadian clock by dexamethasone treatment, the phagocytic activity of normal ARPE-19 cells exhibited significant 24-h oscillation, but this oscillation was modulated by senescence. The phagocytic activity of senescent ARPE-19 cells increased constantly throughout the 24-h period, which still exhibited blunted circadian oscillation, accompanied by an alteration in the rhythmic expression of circadian clock genes and clock-controlled phagocytosis-related genes. The expression levels of REV-ERBα, a molecular component of the circadian clock, were constitutively increased in senescent ARPE-19 cells. Furthermore, pharmacological activation of REV-ERBα by its agonist SR9009 enhanced the phagocytic activity of normal ARPE-19 cells and increased the expression of clock-controlled phagocytosis-related genes. Our present findings extend to understand the role of circadian clock in the alteration of phagocytic activity in RPE during aging. Constitutive enhancement of phagocytic activity of senescent RPE may contribute to age-related retinal degeneration.

Assessment of a Small Molecule Synthetic Lignan in Enhancing Oxidative Balance and Decreasing Lipid Accumulation in Human Retinal Pigment Epithelia.

Visual function depends on the intimate structural, functional and metabolic interactions between the retinal pigment epithelium (RPE) and the neural retina. The daily phagocytosis of the photoreceptor outer segment tips by the overlaying RPE provides essential nutrients for the RPE itself and photoreceptors through intricate metabolic synergy. Age-related retinal changes are often characterized by metabolic dysregulation contributing to increased lipid accumulation and peroxidation as well as the release of proinflammatory cytokines. LGM2605 is a synthetic lignan secoisolariciresinol diglucoside (SDG) with free radical scavenging, antioxidant and anti-inflammatory properties demonstrated in diverse in vitro and in vivo inflammatory disease models. In these studies, we tested the hypothesis that LGM2605 may be an attractive small-scale therapeutic that protects RPE against inflammation and restores its metabolic capacity under lipid overload. Using an in vitro model in which loss of the autophagy protein, LC3B, results in defective phagosome degradation and metabolic dysregulation, we show that lipid overload results in increased gasdermin cleavage, IL-1 β release, lipid accumulation and decreased oxidative capacity. The addition of LGM2605 resulted in enhanced mitochondrial capacity, decreased lipid accumulation and amelioration of IL-1 β release in a model of defective lipid homeostasis. Collectively, these studies suggest that lipid overload decreases mitochondrial function and increases the inflammatory response, with LGM2605 acting as a protective agent.

The cell biology of the retinal pigment epithelium.

The retinal pigment epithelium (RPE), a monolayer of post-mitotic polarized epithelial cells, strategically situated between the photoreceptors and the choroid, is the primary caretaker of photoreceptor health and function. Dysfunction of the RPE underlies many inherited and acquired diseases that cause permanent blindness. Decades of research have yielded valuable insight into the cell biology of the RPE. In recent years, new technologies such as live-cell imaging have resulted in major advancement in our understanding of areas such as the daily phagocytosis and clearance of photoreceptor outer segment tips, autophagy, endolysosome function, and the metabolic interplay between the RPE and photoreceptors. In this review, we aim to integrate these studies with an emphasis on appropriate models and techniques to investigate RPE cell biology and metabolism, and discuss how RPE cell biology informs our understanding of retinal disease.

High-Resolution SD-OCT and EDI-OCT in the Evaluation and Management of Multifocal Serpiginoed Choroditis

Purpose To describe spectral domain optical coherence tomography (SD-OCT) and enhanced depth image OCT (EDI-OCT) findings of multifocal serpiginoid choroditis (MSC) , including affected layer of retinal involvement, changes at the vitreoretinal interface, and response to therapy. Methods A retrospective review of 20 eyes (14 patients) with MSC. Each patient underwent a complete ophthalmologic examination, fundus photography, fundus autoflorecence (FAF) and OCT imaging of the affected retina at the initial visit and on each follow-up. Results In acute stage, SD-OCT showed hyperreflective areas involving the outer retinal layers which include retinal pigment epithelium (RPE), photoreceptor outer segment tips (POST), inner segment–outer segment (IS/OS) junction, external limiting membrane (ELM), and outer nuclear layer (ONL) with choroidal and intraretinal layer cells infiltrate. EDI-OCT showed increase choroidal thickness. As the lesions began to heal, irregular, knobby elevations of outer retinal layers appeared (RPE, POST, IS/OS junction, and ELM could not be distinguished) with significant decrease in choroidal and intraretinal cells. On complete healing, loss of RPE, POST, IS/OS junction, and ELM in SD-OCT scan and absent of the choroidal and intraretinal cells and continous hyperreflactivity of the choroid (increased penetrance). Conclusion SD-OCT and EDI-OCT provides high-resolution detail regarding ultrastructural changes in vitreoretinal interface, outer retina and choroid during the course of the lesion. Serial SD-OCT and EDI-OCT also provides further insight into response to therapy by observing choroidal and intraretinal cells.

Localization of Acetylcholine-Related Molecules in the Retina: Implication of the Communication from Photoreceptor to Retinal Pigment Epithelium

It has been long speculated that specific signals are transmitted from photoreceptors to the retinal pigment epithelium (RPE). However, such signals have not been identified. In this study, we examined the retinal expression and localization of acetylcholine-related molecules as putative candidates for these signals. Previous reports revealed that α7 nicotinic acetylcholine receptors (nAChRs) are present in the microvilli of RPE cells that envelope the tips of photoreceptor outer segments (OS). Secreted mammalian leukocyte antigen 6/urokinase-type plasminogen activator receptor-related protein-1 (SLURP-1) is a positive allosteric modulator of the α7 nAChR. Therefore, we first focused on the expression of SLURP-1. SLURP-1 mRNA was expressed in the outer nuclear layer, which is comprised of photoreceptor cell bodies. SLURP-1 immunoreactivity co-localized with rhodopsin and S-opsin in photoreceptor OS, while choline acetyltransferase (ChAT) and high affinity choline transporter (CHT-1) were also expressed in photoreceptor OS. Immunoelectron microscopy identified that the majority of SLURP-1 was localized to the plasma membranes of photoreceptor OS. These results provide evidence that SLURP-1 is synthesized in photoreceptor cell bodies and transported to photoreceptor OS, where SLURP-1 may also be secreted. Our findings suggest that photoreceptor OS communicate via neurotransmitters such as ACh and SLURP-1, while RPE cells might receive these signals through α7 nAChRs in their microvilli.

High-resolution spectral domain optical coherence tomography and fundus autofluorescence correlation in tubercular serpiginouslike choroiditis

ObjectiveThis study aims to describe changes in high-resolution spectral domain optical coherence tomography (SD-OCT) scans with simultaneous fundus autoflorescence (FAF) signals in tubercular serpiginouslike choroiditis (SLC).MethodsSimultaneous SD-OCT and FAF imaging of eyes affected with SLC from acute stage until resolution of lesions was obtained using Spectralis HRA+OCT system (Heidelberg Engineering, Heidelberg, Germany).PatientsFour eyes (three patients) with SLC were prospectively followed.ResultsAcute lesions of SLC (diffusely hyperautofluorescent) corresponded to hyperreflective areas on SD-OCT involving the retinal pigment epithelium (RPE), photoreceptor outer segment tips (POST), inner segment–outer segment (IS/OS) junction, external limiting membrane (ELM), and outer nuclear layer (ONL) with a minimal distortion of inner retinal layers. There was no backscattering from inner choroid. During healing, lesions became discrete with a hypoautofluorescent border and predominant hyperautofluorescence centrally. The hyperreflective fuzzy areas on SD-OCT scans disappeared, and irregular, knobbly elevations of outer retinal layers appeared. The RPE, POST, IS/OS junction, and ELM could not be distinguished. The ONL appeared normal. The choroid showed an increased reflectance. As the lesions healed further over the next 3–6 months, they became predominantly hypoautofluorescent with loss of RPE, POST, IS/OS junction, and ELM in SD-OCT scan.ConclusionThe SD-OCT provided an insight into the ultrastructural changes in the outer retina during the course of acute SLC lesions. The changes on OCT correlated with abnormal FAF findings.

Melanosomes at a glance

Melanosomes, the pigment granules that provide tissues with colour and photoprotection, are the cellular site of synthesis, storage and transport of melanin pigments. They are synthesised in mammalian skin melanocytes, in choroidal melanocytes and retinal pigment epithelial (RPE) cells in the eye,

Characterization of Outer Retinal Morphology with High-Speed, Ultrahigh-Resolution Optical Coherence Tomography

To visualize, quantitatively assess, and interpret outer retinal morphology by using high-speed, ultrahigh-resolution (UHR) OCT. Retinal imaging was performed in the ophthalmic clinic in a cross-section of 43 normal subjects with a 3.5-microm, axial-resolution, high-speed, UHR OCT prototype instrument, using a radial scan pattern (24 images, 1500 axial scans). Outer retinal layers were automatically segmented and measured. High-definition imaging was performed with a 2.8-microm axial-resolution, high-speed, UHR OCT research prototype instrument, to visualize the finer features in the outer retina. Quantitative maps of outer retinal layers showed clear differences between the cone-dominated fovea and the rod-dominated parafovea and perifovea, indicating that photoreceptor morphology can explain the appearance of the outer retina in high-speed, UHR OCT images. Finer, scattering bands were visualized in the outer retina using high-definition imaging and were interpreted by comparison to known anatomy. High-speed UHR OCT enables quantification of scattering layers in the outer retina. An interpretation of these features is presented and supported by quantitative measurements in normal subjects and comparison with known anatomy. The thick scattering region of the outer retina previously attributed to the retinal pigment epithelium (RPE) is shown to consist of distinct scattering bands corresponding to the photoreceptor outer segment tips, RPE, and Bruch's membrane. These results may advance understanding of the outer retinal appearance in OCT images. The normative measurements may also aid in future investigations of outer retinal changes in age-related macular degeneration and other diseases.

The phagocytosis of os is mediated by the PI3-kinase linked tyrosine kinase receptor, mer, and is stimulated by GAS6.

The phagocytosis of the shed tips of photoreceptor outer segments (OS) by the cells of the adjacent retinal pigment epithelium (RPE) is vital to the survival of the photoreceptor (PR) cells (Young and Bok, 1969). If this phagocytosis is inhibited, death of the PRs occurs. This situation is observed in the Royal College of Surgeons (RCS) strain of rat (Bourne et al., 1938; Dowling and Sidman, 1962). In this animal model, a recessive mutation in the receptor tyrosine kinase (RTK) genemercauses the introduction of a stop codon early in the reading frame, with the result that functional Mer protein is never produced by the RPE cells (D’Cruz et al., 2000; Nandrot et al., 2000). This mutation affects the ability of the RPE cells to perform their normal function of phagocytosing the shed tips of the OS, resulting in degeneration of the underlying photoreceptor cells, and blindness (Bok and Hall, 1971; Mullen and LaVail, 1976; Edwards and Szamier, 1977).

The role of the retinal pigment epithelium: topographical variation and ageing changes.

The retinal pigment epithelium (RPE) is a single layer of post-mitotic cells, which functions both as a selective barrier to and a vegetative regulator of the overlying photoreceptor layer, thereby playing a key role in its maintenance. Through the expression and activity of specific proteins, it regulates the transport of nutrients and waste products to and from the retina, it contributes to outer segment renewal by ingesting and degrading the spent tips of photoreceptor outer segments, it protects the outer retina from excessive high-energy light and light-generated oxygen reactive species and maintains retinal homeostasis through the release of diffusible factors. The ageing characteristics of the RPE suggest that in addition to cell loss, pleomorphic changes and loss of intact melanin granules, significant metabolic changes occur resulting, at least in part, from the intracellular accumulation of lipofuscin. This pigment has been shown to be highly phototoxic and has been linked to several oxidative changes, some leading to cell death. While the aetiology of age-related macular degeneration is complex and is as yet unresolved, it is likely that accelerated ageing-like changes in the RPE play a fundamental role in the development of this condition.

Photopigment transmittance imaging of the primate photoreceptor mosaic

We introduce a new technique for classifying many photoreceptors simultaneously in fresh, excised primate retina on the basis of their absorptance spectra. Primate retina is removed from the pigment epithelium and illuminated under a microscope from the same direction as in the intact eye. To facilitate the guiding of light into the receptor outer segments, the optical axes of the photoreceptors are oriented parallel to the optical axis of the microscope. Photoreceptor outer-segment tips are imaged on a charge-coupled device array, which provides radiometric measurements of the light passing through each photoreceptor. These images are acquired sequentially at three wavelengths chosen to maximize the absorptance differences among the three cone photopigments. After the photopigment is bleached, a second set of three images is acquired. The ratios of the images before and after bleaching at each wavelength are photopigment transmittance maps of the retina. These are combined into a single trichromatic image showing the distribution of photopigment if the retina could be viewed directly in white light without bleaching. We have found patches of receptors in peripheral macaque retina where the measured absorptance at the wavelength of maximum absorptance is consistent with the predicted axial absorptance of th photopigment. The cones in these patches cluster into two groups corresponding to the middle wavelength-sensitive (n = 53, mean absorptance = 0.28) and the long wavelength-sensitive (n = 63, mean absorptance = 0.30) cones. The mean absorptances of 273 macaque and 183 human rods were 0.51 and 0.41, respectively.

Phagocytosis of outer segments by retinal pigment epithelium: phagosome-lysosome interaction.

We studied phagocytosis of rod outer segments (ROS) by the retinal pigmented epithelium (RPE) using rapid freezing, freeze-drying, and electron microscopic immunocytochemistry. Phagocytosis of photoreceptor outer segment tips by the RPE occurs daily, and in rats the shedding of these tips is light-entrained to a circadian rhythm. We studied the phagocytic process 5, 30, 90, and 150 min after light onset or after subjective light onset in rats entrained to a 12-hr dark-12-hr light cycle. Lysosomes were labeled with antibodies to cathepsin D, a major lysosomal enzyme responsible for opsin degradation. Phagosomes and phagolysosomes were recognized because of the lamellar structure of their photoreceptor-derived contents. We found a population of lysosomes that fuse with one another before they interact with phagosomes. This fusion can be triggered either by light or by endogenous circadian mechanisms. We also found that lysosome-phagosome interaction occurs after the ingestion stage is completed and that this interaction occurs in two steps. First, smaller lysosomes fuse with phagosomes. Subsequently, larger lysosomes appear to interact with phagosomes via pore-like or bridge-like structures. It is proposed that interchange of contents takes place through these structures.

Diurnal variation in myeloid bodies of the chick retinal pigment epithelium

Myeloid bodies (MBs) are distinct lamellar regions of the normally branched tubular smooth endoplasmic reticulum of retinal pigment epithelial (RPE) cells. These organelles are present in the retinas of many lower vertebrates. Previous investigations have reported a relationship between outer segment disk shedding and phagocytosis by the RPE, and the formation of MBs. The current morphometric study was undertaken to establish if a temporal relationship existed between MB occurrence and the phagocytosis of shed photoreceptor outer segment tips in the chick retina. We report on the occurrence of phagosomes, MBs, and for the first time, MB precursors (templates) in the RPE over a 24-hr diurnal cycle. Phagosome numbers were observed to be highest within 2 hr following lights on, and again following lights off, while MB precursors were most prevalent at two time points, immediately prior to the times for rod and cone outer segment shedding at lights on and lights off respectively. Traditional MBs of the small (0.06 to 0.239 microns2) and medium (0.24 to 0.99 microns2) variety increased both in size and number from 01:00 hr of a 24 hr time period, with smaller and fewer MBs being present late in the dark part of the diurnal cycle, and larger and more numerous MBs in the later part of the light portion and into the early part of the dark portion of the cycle.(ABSTRACT TRUNCATED AT 250 WORDS)

Photoreceptor shedding is initiated by light in the frog retina.

Frogs maintained on a diurnal light-dark cycle (14 hours light and 10 hours darkness) shed their rod photoreceptor outer segment tips shortly after the onset of light. Shedding is synchronous and occurs in about 25 percent of the rod photoreceptors each day. Prolonged exposure to total darkness decreases the amount of shedding, after which exposure to light results in a large burst of synchronous shedding. Thus in the frog retina, the synchronous shedding of rod outer segment tips is shown to be directly related to light stimulation.