Photoreceptor Level Research Articles

Docosahexaenoic Acid (DHA) and Eicosapentaenoic Acid (EPA)-Should They Be Mandatory Supplements in Pregnancy?

Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) are essential fatty acids for the human body. Seafood and microalgae are the most important sources of omega-3 fatty acids. Supplementation with 200 mg/day of DHA during pregnancy and breastfeeding has been suggested for women and infants in countries with low seafood consumption. Maternal concentration of DHA and EPA was associated with concentration in cord blood and breast milk. High concentrations of DHA and EPA were identified at the level of retinal photoreceptors and neuronal cell membranes. It was observed that supplementation with DHA and EPA during pregnancy had beneficial effects on the neurological development of the fetus and infant by improving language, memory, attention, and hand coordination, affecting sleep patterns, and improving visual acuity. Beneficial effects on the development of the infant were also associated with the maternal intake of omega-3 fatty acids during breastfeeding. Supplementation with DHA and EPA may reduce the risk of preterm birth but also of preeclampsia in low-risk pregnancies. Women of childbearing age should have an intake of 250 mg/day of DHA + EPA from their diet or supplements. To reduce the risk of premature birth, pregnant women must additionally receive at least 100-200 mg of DHA every day. It is recommended that supplementation with omega-3 fatty acids starts before 20 weeks of pregnancy. Beneficial effects on the mother have been identified, such as the reduction of postpartum depression symptoms, the decrease of cardiovascular risk, and the anti-inflammatory role.

Boosting 2-photon vision with adaptive optics.

The 2-photon effect in vision occurs when two photons of the same wavelength are absorbed by cone photopigment in the retina and create a visual sensation matching the appearance of light close to half their wavelength. This effect is especially salient for infrared light, where humans are mostly insensitive to 1-photon isomerizations and thus any perception is dominated by 2-photon isomerizations. This phenomenon can be made more readily visible using short-pulsed lasers, which increase the likelihood of 2-photon excitation by making photon arrivals at the retina more concentrated in time. Adaptive optics provides another avenue for enhancing the 2-photon effect by focusing light more tightly at the retina, thereby increasing the spatial concentration of incident photons. This article makes three contributions. First, we demonstrate through color-matching experiments that an adaptive optics correction can provide a 25-fold increase in the luminance of the 2-photon effect-a boost equivalent to reducing pulse width by 96%. Second, we provide image-based evidence that the 2-photon effect occurs at the photoreceptor level. Third, we use our results to compute the specifications for a system that could utilize 2-photon vision and adaptive optics to image and stimulate the retina using a single infrared wavelength and reach luminance levels comparable to conventional displays.

Effect of Trace Metal Ions on the Conformational Stability of the Visual Photoreceptor Rhodopsin.

Trace metals are essential elements that play key roles in a number of biochemical processes governing human visual physiology in health and disease. Several trace metals, such as zinc, have been shown to play important roles in the visual phototransduction process. In spite of this, there has been little research conducted on the direct effect of trace metal elements on the visual photoreceptor rhodopsin. In the current study, we have determined the effect of several metal ions, such as iron, copper, chromium, manganese, and nickel, on the conformational stability of rhodopsin. To this aim, we analyzed, by means of UV-visible and fluorescence spectroscopic methods, the effects of these trace elements on the thermal stability of dark rhodopsin, the stability of its active Metarhodopsin II conformation, and its chromophore regeneration. Our results show that copper prevented rhodopsin regeneration and slowed down the retinal release process after illumination. In turn, Fe3+, but not Fe2+, increased the thermal stability of the dark inactive conformation of rhodopsin, whereas copper ions markedly decreased it. These findings stress the important role of trace metals in retinal physiology at the photoreceptor level and may be useful for the development of novel therapeutic strategies to treat retinal disease.

Features of Visual Dysfunctions in Patients with Primary Hypothyroidism and Thyrotoxicosis

Purpose: to study the prevalence and nature of visual dysfunctions in patients with primary hypothyroidism and thyrotoxicosis.Material and methods. The material for this study was the results of a survey of 54 patients (108 eyes) with thyroid dysfunctions: 32 people (64 eyes) with primary untreated hypothyroidism and 22 people (44 eyes) with primary untreated thyrotoxicosis. Static automated perimetry and dedicated shortwave infrared (blue-yellow) perimetry were performed. The average total value of the photosensitivity of each (n = 74) tested point of the field of view was analyzed, the topography of the location of focal defects was studied, and the severity of impairments to photosensitivity was assessed by aggregate signs.Results. Reliably high sensitivity (92.6 %) and specificity (50.0 %) of short-wave infrared perimetry in relation to static automated perimetry were revealed. In thyroid dysfunctions, the prevalence of optic neuropathy reaches 93 % according to the data of short-wave infrared perimetry versus 7 % of static automated perimetry. It is manifested by a diffuse decrease in light sensitivity to blue stimulus with an increase in the depth of depression from the center to the periphery in both types of thyroid dysfunction. Against this background, with primary hypothyroidism, focal defects appeared in the form of first-order scotomas, and with primary thyrotoxicosis, second-order scotomas. Scotomas were located at the periphery of the central visual field, 20–30° from the fixation point. In the analyzed groups, high average group indices of the maximum corrected visual acuity were established, which allows us to speak about the safety of the photopic (cone) component of the visual analyzer.Conclusion. The pattern of photosensitivity disorders, the topography of the location of the loci of local defects revealed by the short-wave infrared perimetry, indicate that the earliest signs of optical neuropathy are manifested at the level of photoreceptors — selectively in the S-cones. Decreased sensitivity to blue stimulus (440 nm) refers to an acquired color anomaly called tritanopia; which can be present with high visual function, is most often associated with a decrease in the number of S-cones and a deficiency of retinol (the source of cyanolab synthesis).

Structural basis of retinal membrane guanylate cyclase regulation by GCAP1 and RD3.

Retinal membrane guanylate cyclases (RetGC1 and RetGC2) are expressed in photoreceptor rod and cone cells, where they promote the onset of visual recovery during phototransduction. The catalytic activity of RetGCs is regulated by their binding to regulatory proteins, guanylate cyclase activating proteins (GCAP1-5) and the retinal degeneration 3 protein (RD3). RetGC1 is activated by its binding to Ca2+-free/Mg2+-bound GCAP1 at low cytosolic Ca2+ levels in light-activated photoreceptors. By contrast, RetGC1 is inactivated by its binding to Ca2+-bound GCAP1 and/or RD3 at elevated Ca2+ levels in dark-adapted photoreceptors. The Ca2+ sensitive cyclase activation helps to replenish the cytosolic cGMP levels in photoreceptors during visual recovery. Mutations in RetGC1, GCAP1 or RD3 that disable the Ca2+-dependent regulation of cyclase activity are genetically linked to rod/cone dystrophies and other inherited forms of blindness. Here I review the structural interaction of RetGC1 with GCAP1 and RD3. I propose a two-state concerted model in which the dimeric RetGC1 allosterically switches between active and inactive conformational states with distinct quaternary structures that are oppositely stabilized by the binding of GCAP1 and RD3. The binding of Ca2+-free/Mg2+-bound GCAP1 is proposed to activate the cyclase by stabilizing RetGC1 in an active conformation (R-state), whereas Ca2+-bound GCAP1 and/or RD3 inhibit the cyclase by locking RetGC1 in an inactive conformation (T-state). Exposed hydrophobic residues in GCAP1 (residues H19, Y22, M26, F73, V77, W94) are essential for cyclase activation and could be targeted by rational drug design for the possible treatment of rod/cone dystrophies.

NMR Structures of Calmodulin Bound to Two Separate Regulatory Sites in the Retinal Cyclic Nucleotide-Gated Channel.

Retinal cyclic nucleotide-gated (CNG) channels (composed of three CNGA1 and one CNGB1 subunits) exhibit a Ca2+-induced reduction in channel open probability mediated by calmodulin (CaM). Defects in the Ca2+-dependent regulation of CNG channels may be linked to autosomal recessive retinitis pigmentosa and other inherited forms of blindness. Here, we report the NMR structure and binding analysis of CaM bound to two separate sites within CNGB1 (CaM1: residues 565-589 and CaM2: residues 1120-1147). Our binding studies reveal that CaM1 binds to the Ca2+-bound CaM N-lobe with at least fivefold higher affinity than it binds to the CaM C-lobe. By contrast, the CaM2 site binds to the Ca2+-bound CaM C-lobe with higher affinity than it binds to the N-lobe. CaM1 and CaM2 both exhibited very weak binding to Ca2+-free CaM. We present separate NMR structures of Ca2+-saturated CaM bound to CaM1 and CaM2 that define key intermolecular contacts: CaM1 residue F575 interacts with the CaM N-lobe while CaM2 residues L1129, L1132, and L1136 each make close contact with the CaM C-lobe. The CNGB1 mutation F575E abolishes CaM1 binding to the CaM N-lobe while L1132E and L1136E each abolish CaM2 binding to the CaM C-lobe. Thus, a single CaM can bind to both sites in CNGB1 in which the CaM N-lobe binds to CaM1 and the CaM C-lobe binds to CaM2. We propose a Ca2+-dependent conformational switch in the CNG channel caused by CaM binding, which may serve to attenuate cGMP binding to CNG channels at high cytosolic Ca2+ levels in dark-adapted photoreceptors.

Chemical shift assignments of calmodulin bound to a C-terminal site (residues 1120–1147) in the β-subunit of a retinal cyclic nucleotide-gated channel (CNGB1)

Retinal cyclic nucleotide-gated (CNG) channels consist of two protein subunits (CNGA1 and CNGB1). Calmodulin (CaM) binds to two separate sites within the cytosolic region of CNGB1: CaM binding to an N-terminal site (human CNGB1 residues 565–587, called CaM1) decreases the open probability of CNG channels at elevated Ca2+ levels in dark-adapted photoreceptors, whereas CaM binding to a separate C-terminal site (CNGB1 residues 1120–1147, called CaM2) may increase channel open probability in light activated photoreceptors. We recently reported NMR chemical shift assignments of Ca2+-saturated CaM bound to the CaM1 site of CNGB1 (BMRB no. 51222). Here, we report complete NMR chemical shift assignments of Ca2+-saturated CaM bound to the C-terminal CaM2 site of CNGB1 (BMRB no. 51447).

Kinase activity profiling identifies putative downstream targets of cGMP/PKG signaling in inherited retinal neurodegeneration

Inherited retinal diseases (IRDs) are a group of neurodegenerative disorders that lead to photoreceptor cell death and eventually blindness. IRDs are characterised by a high genetic heterogeneity, making it imperative to design mutation-independent therapies. Mutations in a number of IRD disease genes have been associated with a rise of cyclic 3’,5’-guanosine monophosphate (cGMP) levels in photoreceptors. Accordingly, the cGMP-dependent protein kinase (PKG) has emerged as a new potential target for the mutation-independent treatment of IRDs. However, the substrates of PKG and the downstream degenerative pathways triggered by its activity have yet to be determined. Here, we performed kinome activity profiling of different murine organotypic retinal explant cultures (diseased rd1 and wild-type controls) using multiplex peptide microarrays to identify proteins whose phosphorylation was significantly altered by PKG activity. In addition, we tested the downstream effect of a known PKG inhibitor CN03 in these organotypic retina cultures. Among the PKG substrates were potassium channels belonging to the Kv1 family (KCNA3, KCNA6), cyclic AMP-responsive element-binding protein 1 (CREB1), DNA topoisomerase 2-α (TOP2A), 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (F263), and the glutamate ionotropic receptor kainate 2 (GRIK2). The retinal expression of these PKG targets was further confirmed by immunofluorescence and could be assigned to various neuronal cell types, including photoreceptors, horizontal cells, and ganglion cells. Taken together, this study confirmed the key role of PKG in photoreceptor cell death and identified new downstream targets of cGMP/PKG signalling that will improve the understanding of the degenerative mechanisms underlying IRDs.

Opposing Actions of Octopamine and Tyramine on Honeybee Vision.

The biogenic amines octopamine and tyramine are important neurotransmitters in insects and other protostomes. They play a pivotal role in the sensory responses, learning and memory and social organisation of honeybees. Generally, octopamine and tyramine are believed to fulfil similar roles as their deuterostome counterparts epinephrine and norepinephrine. In some cases opposing functions of both amines have been observed. In this study, we examined the functions of tyramine and octopamine in honeybee responses to light. As a first step, electroretinography was used to analyse the effect of both amines on sensory sensitivity at the photoreceptor level. Here, the maximum receptor response was increased by octopamine and decreased by tyramine. As a second step, phototaxis experiments were performed to quantify the behavioural responses to light following treatment with either amine. Octopamine increased the walking speed towards different light sources while tyramine decreased it. This was independent of locomotor activity. Our results indicate that tyramine and octopamine act as functional opposites in processing responses to light.

Structural Insights into Retinal Guanylate Cyclase Activator Proteins (GCAPs).

Retinal guanylate cyclases (RetGCs) promote the Ca2+-dependent synthesis of cGMP that coordinates the recovery phase of visual phototransduction in retinal rods and cones. The Ca2+-sensitive activation of RetGCs is controlled by a family of photoreceptor Ca2+ binding proteins known as guanylate cyclase activator proteins (GCAPs). The Mg2+-bound/Ca2+-free GCAPs bind to RetGCs and activate cGMP synthesis (cyclase activity) at low cytosolic Ca2+ levels in light-activated photoreceptors. By contrast, Ca2+-bound GCAPs bind to RetGCs and inactivate cyclase activity at high cytosolic Ca2+ levels found in dark-adapted photoreceptors. Mutations in both RetGCs and GCAPs that disrupt the Ca2+-dependent cyclase activity are genetically linked to various retinal diseases known as cone-rod dystrophies. In this review, I will provide an overview of the known atomic-level structures of various GCAP proteins to understand how protein dimerization and Ca2+-dependent conformational changes in GCAPs control the cyclase activity of RetGCs. This review will also summarize recent structural studies on a GCAP homolog from zebrafish (GCAP5) that binds to Fe2+ and may serve as a Fe2+ sensor in photoreceptors. The GCAP structures reveal an exposed hydrophobic surface that controls both GCAP1 dimerization and RetGC binding. This exposed site could be targeted by therapeutics designed to inhibit the GCAP1 disease mutants, which may serve to mitigate the onset of retinal cone-rod dystrophies.

Opsin Expression Varies with Reproductive State in the Cichlid Fish Astatotilapia burtoni.

Animals use visual communication to convey crucial information about their identity, reproductive status, and sex. Plasticity in the auditory and olfactory systems has been well-documented, however, fewer studies have tested for plasticity in the visual system, a surprising detail since courtship and mate choice are largely dependent on visual signals across taxa. We previously found reproductive state-dependent plasticity in the eye of the highly social cichlid fish Astatotilapia burtoni. Male A. burtoni increase their courtship, including multicomponent visual displays, when around ovulated females, and ovulated females are more responsive to male visual courtship displays than non-ovulated females. Based on this, we hypothesized that ovulation status impacts visual capabilities in A. burtoni females. Using electroretinograms, we found that ovulated females had greater visual sensitivity at wavelengths corresponding to male courtship coloration compared with non-reproductively-receptive females. In addition, ovulated females had higher neural activation in the retina and higher mRNA expression levels of neuromodulatory receptors (e.g., sex-steroids; gonadotropins) in the eye than non-ovulated females. Here, we add to this body of work by testing the hypothesis that cone opsin expression changes with female reproductive state. Ovulated females had higher expression of short wavelength sensitive opsins (sws1, sws2a, sws2b) compared with mouthbrooding females. Further, expression of sws2a, the most abundant opsin in the A. burtoni eye, positively correlated with levels of circulating 11-ketotestosterone and estradiol and estrogen, androgen, and gonadotropin system receptor expression in the eye in females. These data indicate that reproductive state-dependent plasticity also occurs at the level of photoreceptors, not just through modulation of visual signals at downstream retinal layers. Collectively, these data provide crucial evidence linking endocrine modulation of visual plasticity to mate choice behaviors in females.

Color discrimination properties arising from optimal decoding in the early stages of visual systems

In order to interpret animal behavior we need to understand how they see the world. As directly testing color discrimination in animals is difficult and time consuming, it is important to develop theoretical models based in the properties of visual systems. One of the most successful for the prediction of color discrimination behavior is the receptor noise-limited (RNL) model, which depends only on the level of noise in photoreceptors and opponent mechanisms. Here a complementary approach to model construction is used, and optimal color discrimination properties are obtained using information theoretical tools, for the early stages of visual systems. It is shown here that, for most biologically relevant conditions the optimal discrimination function of an ideal observer coincides with the one obtained with the RNL model. Furthermore, within this framework the influence of opponency can be studied by considering models with and without that mechanism but with exactly the same parameters at the level of photoreceptors. As an example, it is shown here that opponency is necessary to explain the discrimination of monochromatic stimuli in honeybees, but not in budgerigars. Since this is a consequence of the narrowing of absorption spectra of photoreceptors, produced by the presence of oil droplets, this could also be true for most other species of birds. This suggests that in order to study opponency in birds, stimuli should have a relatively wide spectrum.

Mapping membrane lipids in the developing and adult mouse retina under physiological and pathological conditions using mass spectrometry

Membrane phospholipids play pivotal roles in various cellular processes, and their levels are tightly regulated. In the retina, phospholipids had been scrutinized because of their distinct composition and requirement in visual transduction. However, how lipid composition changes during retinal development remains unclear. Here, we used liquid chromatography–mass spectrometry (LC-MS) to assess the dynamic changes in the levels of two main glycerophospholipids, phosphatidylcholine (PC) and phosphatidylethanolamine (PE), in the developing mouse retina under physiological and pathological conditions. The total levels of PC and PE increased during retinal development, and individual lipid species exhibited distinct level changes. The amount of very-long-chain PC and PE increased dramatically in the late stages of retinal development. The mRNA levels of Elovl2 and Elovl4, genes encoding enzymes essential for the synthesis of very-long-chain polyunsaturated fatty acids, increased in developing photoreceptors. Cell sorting based on CD73 expression followed by LC-MS revealed distinct changes in PC and PE levels in CD73-positive rod photoreceptors and CD73-negative retinal cells. Finally, using the NaIO3-induced photoreceptor degeneration model, we identified photoreceptor-specific changes in PC and PE levels from 1 day after NaIO3 administration, before the outer segment of photoreceptors displayed morphological impairment. In conclusion, our findings provide insight into the dynamic changes in PC and PE levels in the developing and adult mouse retina under physiological and pathological conditions. Furthermore, we provide evidence that cell sorting followed by LC-MS is a promising approach for investigating the relevance of lipid homeostasis in the function of different retinal cell types.

Light Adaptation of Retinal Rods, Adaptation Memory, and Afterimages

The GPCR signal cascade in retinal photoreceptor cells – rods and cones – supports powerful signal amplification and allows rods to respond reliably to single light quanta. Rods remain functional at light fluxes down to 105 quanta/sec, which is supported by the highly effective light adaptation system. Light adaptation is based on negative feedback circuits, mostly via changes in intracellular Ca2+ ion concentrations. Three calcium regulation loops have been reliably identified – acceleration of quenching of light-activated rhodopsin, acceleration of cGMP synthesis by guanylate cyclase, and increased affinity of cGMP-controlled ion cannels for nucleotide. However, one or two further highly effective mechanisms of adaptation are known to operate, one of which regulates the lifetime of activated phosphodiesterase; for others, the target of regulation is unknown. The mediators of this regulation are also unknown. Our studies of these mechanisms revealed a novel phenomenon, which has been missed in previous studies. We found that recovery of the dark current of rods after cessation of non-saturating adapting light can take 20–30 sec. Furthermore, after formal return of the membrane current to the dark level, cell sensitivity to test stimuli remains decreased for a further 1–2 min. We termed this phenomenon “adaptation memory.” Adaptation memory is reminiscent of the phenomenology of afterimages. The gradual return of the membrane current to the dark level may correspond to fading of the positive afterimage. Prolonged decreases in photoreceptor sensitivity to additional stimulation can create a negative afterimage. As far as we know, this is the first experimental physiological demonstration of the ability to generate afterimages at the level of single photoreceptors.

Effect of AAV-Mediated Rhodopsin Gene Augmentation on Retinal Degeneration Caused by the Dominant P23H Rhodopsin Mutation in a Knock-In Murine Model.

Mutations in the rhodopsin gene may cause photoreceptor degeneration in autosomal dominant retinitis pigmentosa (ADRP) by dominant negative or toxic gain-of-function mechanisms. Controversy exists as to the mechanism by which the widely studied P23H mutation induces rod cell dysfunction and death. Inherited disease caused by dominant negative mutations may be amenable to treatment using wild-type gene augmentation. Indeed, prior studies in the RHOP23H, Rho+/- transgenic mouse model of ADRP have suggested that a therapeutic benefit may be achieved when wild-type rhodopsin is overexpressed following subretinal delivery of a recombinant adeno-associated viral (AAV) vector. In this study, we investigated the effect of wild-type rhodopsin supplementation on the rate of retinal degeneration in the more clinically relevant RhoP23H/+ knock-in mouse model of ADRP. Four AAVs carrying the human rhodopsin coding sequence were first designed and compared for efficacy in the rhodopsin knockout mouse. All four vectors were capable of driving expression of the human transgene in the knockout retina with the protein being appropriately trafficked to de novo rod outer segments. The most efficient of these vectors was injected at one of two doses into the subretinal space of RhoP23H/+ mice and the effect on retinal structure and function determined longitudinally by spectral-domain optical coherence tomography and electroretinography, respectively, over a 3-month period. Although significant overexpression of rhodopsin protein was achieved in this model, no beneficial effect on retinal structure or function was observed at either dose. Lack of therapeutic efficacy in this model may be attributable to the relative rapidity of degeneration in the RhoP23H/+ mouse relative to the human disease, over- or under dosing at the level of individual photoreceptors, late timing of the intervention, or a possible predominant toxic gain-of-function mechanism of degeneration.

Anterior visual system imaging to investigate energy failure in multiple sclerosis.

Mitochondrial failure and hypoxia are key contributors to multiple sclerosis pathophysiology. Importantly, improving mitochondrial function holds promise as a new therapeutic strategy in multiple sclerosis. Currently, studying mitochondrial changes in multiple sclerosis is hampered by a paucity of non-invasive techniques to investigate mitochondrial function of the CNS in vivo. It is against this backdrop that the anterior visual system provides new avenues for monitoring of mitochondrial changes. The retina and optic nerve are among the metabolically most active structures in the human body and are almost always affected to some degree in multiple sclerosis. Here, we provide an update on emerging technologies that have the potential to indirectly monitor changes of metabolism and mitochondrial function. We report on the promising work with optical coherence tomography, showing structural changes in outer retinal mitochondrial signal bands, and with optical coherence angiography, quantifying retinal perfusion at the microcapillary level. We show that adaptive optics scanning laser ophthalmoscopy can visualize live perfusion through microcapillaries and structural changes at the level of single photoreceptors and neurons. Advantages and limitations of these techniques are summarized with regard to future research into the pathology of the disease and as trial outcome measures.

Световая адаптация палочек сетчатки, адаптационная память и последовательные образы

The GPCR signaling cascade in the retinal photoreceptor cells, rods and cones, provides high signal amplification and allows rods to respond reliably to single photons. At the same time, rods remain operational under light fluxes up to 105 photons per second, which is supported by a highly efficient system of light adaptation. Light adaptation is based on negative feedback chains, mainly through changes in the intracellular concentration of Ca2+ ions. There are three reliably identified loops of calcium regulation – the acceleration of quenching of photoactivated rhodopsin, the acceleration of cGMP synthesis by guanylate cyclase, and an increase of cGMP-controlled ion channels’ affinity to the nucleotide. However, it is known that there are additional one or two highly effective adaptation mechanisms, one of which regulates the lifetime of activated phosphodiesterase; for the second one, the target of regulation is unknown. The messengers of these regulations also remain unknown. While studying these mechanisms, we discovered a new phenomenon that was not noticed in previous works. We found that the restoration of rod’s dark current after turning off the non-saturating adapting light can take 20-30 s. Moreover, after the membrane current formally returns to the dark level, the sensitivity of the cell to testing stimuli remains reduced for one to two minutes. We called this phenomenon “adaptation memory”. Adaptation memory resembles the phenomenology of the afterimages. The gradual return of the membrane current to the dark level could correspond to the fading of positive afterimage. Long-term decrease in the sensitivity of photoreceptors to an additional stimulation can create a negative afterimage. As far as we know, this is the first experimental physiological demonstration of the possibility of afterimages generation already at the level of single photoreceptors.

Photoreceptors in a mouse model of Leigh syndrome are capable of normal light-evoked signaling

Mitochondrial dysfunction is an important cause of heritable vision loss. Mutations affecting mitochondrial bioenergetics may lead to isolated vision loss or life-threatening systemic disease, depending on a mutation's severity. Primary optic nerve atrophy resulting from death of retinal ganglion cells is the most prominent ocular manifestation of mitochondrial disease. However, dysfunction of other retinal cell types has also been described, sometimes leading to a loss of photoreceptors and retinal pigment epithelium that manifests clinically as pigmentary retinopathy. A popular mouse model of mitochondrial disease that lacks NADH:ubiquinone oxidoreductase subunit S4 (NDUFS4), a subunit of mitochondrial complex I, phenocopies many traits of the human disease Leigh syndrome, including the development of optic atrophy. It has also been reported that ndufs4-/- mice display diminished light responses at the level of photoreceptors or bipolar cells. By conducting electroretinography (ERG) recordings in live ndufs4-/- mice, we now demonstrate that this defect occurs at the level of retinal photoreceptors. We found that this deficit does not arise from retinal developmental anomalies, photoreceptor degeneration, or impaired regeneration of visual pigment. Strikingly, the impairment of ndufs4-/- photoreceptor function was not observed in ex vivo ERG recordings from isolated retinas, indicating that photoreceptors with complex I deficiency are intrinsically capable of normal signaling. The difference in electrophysiological phenotypes in vivo and ex vivo suggests that the energy deprivation associated with severe mitochondrial impairment in the outer retina renders ndufs4-/- photoreceptors unable to maintain the homeostatic conditions required to operate at their normal capacity.

Computational simulation of human perception of spatially dependent patterns modulated by degree and angle of linear polarization.

Recent studies on polarization perception have shown that humans are sensitive to patterned stimuli modulated by either angle of linear polarization (AoP) or degree of polarization (DoP). Here, we present a model of human polarization sensitivity that incorporates both AoP and DoP as spatially dependent input variables. Applying the model to both sinusoidal- and square-wave-modulated DoP and AoP inputs, we demonstrate the theoretical similarities and differences generated by such inputs. Our model indicates the following: (i)edge boundaries between two adjacent areas of different linear polarization are preserved for both AoP- and DoP-modulated stimuli; and (ii)compared with DoP stimuli, AoP stimuli generate greater luminance changes at the photoreceptor level, suggesting that AoP-modulated patterns are potentially more salient than DoP patterns. The computational model is supported experimentally with an optical test of the model comprising a radial diattenuating polarizing filter and modified liquid crystal displays generating DoP- and AoP-modulated outputs. Psychophysical measures of human sensitivity confirm the increased salience of AoP- relative to DoP-modulated stimuli. These findings have practical application to the selection of DoP- and AoP-modulated stimuli for the investigation of macular function and macular pigment density in healthy and diseased eyes.

Choroidal melanoma and retinal changes

To study the capabilities of optical coherence tomography (OCT) in terms of finding the ways CM spreads into the retina based on pathomorphological examination of the eyes enucleated due to CM. OCT tomograms of 64 eyes with CM (mean tumor elevation - 2.89±0.19 mm, diameter - 9.58±0.43 mm) were studied. Pathomorphological study was conducted on 18 eyes with CM elevated by up to 7 mm and without previous treatment. OCT revealed signs of melanoma invading into the retina in 49 eyes. Among those signs were: presence of retina/CM aggregation (21 eyes), tearing of Bruch's membrane and retinal pigment epithelium, disturbed differentiation of outer retinal layers (15 eyes). Combination of two types of invasion was found in 11 eyes. Pathomorphological study showed low mitotic activity of CM and the presence of a small number of fine-local necroses and hemorrhages. Invasion of CM into the retina was determined in 13 eyes. Retinal invasion began with local destruction of Bruch's membrane (BM) and was often limited by the photoreceptor level, disturbing architectonics of the retina and nuclear layers. Large CMs were accompanied by rough destruction of the Bruch's membrane, tumor cells - in the edematous retina with cystoid dystrophy, in its inner layers, invading into the vitreous body. Infiltration of the retina to inner nuclear layer was accompanied by its transudative detachment and cystoid dystrophy. Invasion of CM into the retina affecting all of its layers imposes use of OCT to examine the integrity of the retina, even with a tumor of small thickness. Tumoral infiltration of the retina and localization of tumor near the optic disc are two contraindications for local endovitreal resection. Brachytherapy requires consideration of tumor characteristics and thickness of the retina for calculation of apical radiation dose.