Retinal Projections Research Articles

Wiring the Binocular Visual Pathways.

Retinal ganglion cells (RGCs) extend axons out of the retina to transmit visual information to the brain. These connections are established during development through the navigation of RGC axons along a relatively long, stereotypical pathway. RGC axons exit the eye at the optic disc and extend along the optic nerves to the ventral midline of the brain, where the two nerves meet to form the optic chiasm. In animals with binocular vision, the axons face a choice at the optic chiasm—to cross the midline and project to targets on the contralateral side of the brain, or avoid crossing the midline and project to ipsilateral brain targets. Ipsilaterally and contralaterally projecting RGCs originate in disparate regions of the retina that relate to the extent of binocular overlap in the visual field. In humans virtually all RGC axons originating in temporal retina project ipsilaterally, whereas in mice, ipsilaterally projecting RGCs are confined to the peripheral ventrotemporal retina. This review will discuss recent advances in our understanding of the mechanisms regulating specification of ipsilateral versus contralateral RGCs, and the differential guidance of their axons at the optic chiasm. Recent insights into the establishment of congruent topographic maps in both brain hemispheres also will be discussed.

P‐92: A Lensless Retinal Scanning Display for Augmented Reality

We propose a design of a retinal‐scanning‐based near‐eye display for augmented reality. Our solution is highlighted by a scanning laser projector, a diffractive optical element, and a wet eye with gradient refractive indices. Its design rules are set forth in detail, followed by the results and discussion.

Augmented reality technology selection based on integrated QFD-AHP model

In the last decade, Augmented Reality has become increasingly popular. As improved performances are gathered in terms of mature hardware and software tools, we are observing the stemming of a huge number of applications of this technology both in the entertainment and in the industrial domains. On the one hand, such applications are usually claimed to bring benefits in terms of productivity or enhancement of the human’s capability to perform tasks. On the other hand, researchers and developers seem not to adequately consider the different meanings that AR assumes when implemented through visualization devices that can differ significantly in nature and in their capability to provide a mixed real-virtual scenario. In this paper, we describe a user-centred method based on an integrated QFD-AHP approach to select the best visualization display technology with regard to a specific application context. The aim is to establish a repeatable and documented process for the identification of the technology that best suits and mitigates the acceptability risks of the transition from a legacy working environment to an AR based operational environment. The method has been developed in the framework of the RETINA (Resilient Synthetic Vision for Advanced Control Tower Air Navigation Service Provision) project involving the end users, in this case, air traffic controllers. Nevertheless, it can be generalised and applied to other contexts of use. Furthermore, in order to be resilient to the fast, technological development in AR, it can be used to update the results as improvements arise in the performance level of the display devices in a specific technology.

Varifocal Occlusion for Optical See-Through Head-Mounted Displays using a Slide Occlusion Mask.

We propose a varifocal occlusion technique for optical see-through head-mounted displays (OST-HMDs). Occlusion in OST-HMDs is a powerful visual cue that enables depth perception in augmented reality (AR). Without occlusion, virtual objects rendered by an OST-HMD appear semi-transparent and less realistic. A common occlusion technique is to use spatial light modulators (SLMs) to block incoming light rays at each pixel on the SLM selectively. However, most of the existing methods create an occlusion mask only at a single, fixed depth-typically at infinity. With recent advances in varifocal OST-HMDs, such traditional fixed-focus occlusion causes a mismatch in depth between the occlusion mask plane and the virtual object to be occluded, leading to an uncomfortable user experience with blurred occlusion masks. In this paper, we thus propose an OST-HMD system with varifocal occlusion capability: we physically slide a transmissive liquid crystal display (LCD) to optically shift the occlusion plane along the optical path so that the mask appears sharp and aligns to a virtual image at a given depth. Our solution has several benefits over existing varifocal occlusion methods: it is computationally less demanding and, more importantly, it is optically consistent, i.e., when a user loses focus on the corresponding virtual image, the mask again gets blurred consistently as the virtual image does. In the experiment, we build a proof-of-concept varifocal occlusion system implemented with a custom retinal projection display and demonstrate that the system can shift the occlusion plane to depths ranging from 25 cm to infinity.

Focus-free head-mounted display based on Maxwellian view using retroreflector film.

In this study, a focus-free retinal projection display based on a Maxwellian view was implemented. The display was constructed using simple and cost-effective passive components, such as a retroreflector film, beam splitter, and a pinhole. As a part of the system, the pinhole and display serve as a pinhole projector, which projects images through a small exit pupil. The retroreflector film and the beam splitter form the conjugate image of the pinhole near an observer. By locating the center of the crystalline lens of an eye at the conjugate pinhole, the projected images are focused on the retina and are seldomly changed by the accommodative response of the lens. For the practical implementation of the system, a collimating lens was used as an imaging lens combined with a pinhole. The point spread function of the system was derived to theoretically demonstrate that the system exhibits focus-free features at accommodation distances of 0.25, 0.5, and 1m. The experiments conducted to demonstrate the feasibility of the system are presented under the same accommodation condition as the analysis.

Conversations with Ray Guillery on albinism: linking Siamese cat visual pathway connectivity to mouse retinal development.

In albinism of all species, perturbed melanin biosynthesis in the eye leads to foveal hypoplasia, retinal ganglion cell misrouting, and, consequently, altered binocular vision. Here, written before he died, Ray Guillery chronicles his discovery of the aberrant circuitry from eye to brain in the Siamese cat. Ray's characterization of visual pathway anomalies in this temperature sensitive mutation of tyrosinase and thus melanin synthesis in domestic cats opened the exploration of albinism and simultaneously, a genetic approach to the organization of neural circuitry. I follow this account with a remembrance of Ray's influence on my work. Beginning with my postdoc research with Ray on the cat visual pathway, through my own work on the mechanisms of retinal axon guidance in the developing mouse, Ray and I had a continuous and rich dialogue about the albino visual pathway. I will present the questions Ray posed and clues we have to date on the still-elusive link between eye pigment and the proper balance of ipsilateral and contralateral retinal ganglion cell projections to the brain.

Mutant Ahi1 Affects Retinal Axon Projection in Zebrafish via Toxic Gain of Function.

Joubert syndrome (JBTS) is an inherited autosomal recessive disorder associated with cerebellum and brainstem malformation and can be caused by mutations in the Abelson helper integration site-1 (AHI1) gene. Although AHI1 mutations in humans cause abnormal cerebellar development and impaired axonal decussation in JBTS, these phenotypes are not robust or are absent in various mouse models with Ahi1 mutations. AHI1 contains an N-terminal coiled-coil domain, multiple WD40 repeats, and a C-terminal Src homology 3 (SH3) domain, suggesting that AHI1 functions as a signaling or scaffolding protein. Since most AHI1 mutations in humans can result in truncated AHI1 proteins lacking WD40 repeats and the SH3 domain, it remains unclear whether mutant AHI1 elicits toxicity via a gain-of-function mechanism by the truncated AHI1. Because Ahi1 in zebrafish and humans share a similar N-terminal region with a coiled-coil domain that is absent in mouse Ahi1, we used zebrafish as a model to investigate whether Ahi1 mutations could affect axonal decussation. Using in situ hybridization, we found that ahi1 is highly expressed in zebrafish ocular tissues, especially in retina, allowing us to examine its effect on retinal ganglion cell (RGC) projection and eye morphology. We injected a morpholino to zebrafish embryos, which can generate mutant Ahi1 lacking the intact WD40 repeats, and found RGC axon misprojection and ocular dysplasia in 4 dpf (days post-fertilization) larvae after the injection. However, ahi1 null zebrafish showed normal RGC axon projection and ocular morphology. We then used CRISPR/Cas9 to generate truncated ahi1 and also found similar defects in the RGC axon projection as seen in those injected with ahi1 morpholino. Thus, the aberrant retinal axon projection in zebrafish is caused by the presence of mutant ahi1 rather than the loss of ahi1, suggesting that mutant Ahi1 may affect axonal decussation via toxic gain of function.

QD laser eyewear as a visual field aid in a visual field defect model

Visual field defects interfere with free actions and influence the quality of life of patients with retinitis pigmentosa; the prevalence of this disease is increasing in aging societies. Patients with progressive disease may require visual aids; however, no such devices are currently available. We utilized a retinal projection eyewear system, QD laser eyewear, which includes a projector inside the spectacle frame, to draw the image taken by a connected portable camera with a wide field lens. The images are projected onto the retina using a Maxwellian view optical system, which is not influenced by refractive error or the amount of incident light. Goldmann perimetry and figure recognition tests with the QD laser eyewear showed increased visual field areas and angles, and shortened the time for recognition of the number of figures in a sheet, in a limited visual field model that we developed by using a pin-hole system to simulate the tunnel vision of retinitis pigmentosa in 19 healthy adults. The device supported the quality of vision. Additionally, the visual field defect model used in healthy adults was useful for validating the device in the development stage of the study, to clarify both advantages and future goals for improving the device.

Abrogation of Stem Loop Binding Protein (Slbp) function leads to a failure of cells to transition from proliferation to differentiation, retinal coloboma and midline axon guidance deficits.

Through forward genetic screening for mutations affecting visual system development, we identified prominent coloboma and cell-autonomous retinal neuron differentiation, lamination and retinal axon projection defects in eisspalte (ele) mutant zebrafish. Additional axonal deficits were present, most notably at midline axon commissures. Genetic mapping and cloning of the ele mutation showed that the affected gene is slbp, which encodes a conserved RNA stem-loop binding protein involved in replication dependent histone mRNA metabolism. Cells throughout the central nervous system remained in the cell cycle in ele mutant embryos at stages when, and locations where, post-mitotic cells have differentiated in wild-type siblings. Indeed, RNAseq analysis showed down-regulation of many genes associated with neuronal differentiation. This was coincident with changes in the levels and spatial localisation of expression of various genes implicated, for instance, in axon guidance, that likely underlie specific ele phenotypes. These results suggest that many of the cell and tissue specific phenotypes in ele mutant embryos are secondary to altered expression of modules of developmental regulatory genes that characterise, or promote transitions in, cell state and require the correct function of Slbp-dependent histone and chromatin regulatory genes.

CPSF1 mutations are associated with early-onset high myopia and involved in retinal ganglion cell axon projection.

High myopia is a severe form of nearsightedness, which can result in blindness due to its associated complications. While both genetic and environmental factors can cause high myopia, early-onset high myopia (eoHM), which is defined as high myopia that occurs before school age, is considered to be caused mainly by genetic variations, with minimal environmental involvement. Here we report six rare heterozygous loss-of-function (LoF) variants in CPSF1 that were identified in six of 623 probands with eoHM but none of 2657 probands with other forms of genetic eye diseases; this difference was statistically significant (P = 4.60 × 10−5, Fisher’s exact test). The six variants, which were confirmed by Sanger sequencing, were c.3862_3871dup (p.F1291*), c.2823_2824del (p.V943Lfs*65), c.1858C>T (p.Q620*), c.15C>G (p.Y5*), c.3823G>T (p.D1275Y) and c.4146-2A>G. Five of these six variants were absent in existing databases, including gnomAD, 1000G and EVS. The remaining variant, c.4146-2A>G, was present in gnomAD with a frequency of 1/229918. Clinical data demonstrated eoHM in the six probands with these mutations. Knockdown of cpsf1 by morpholino oligonucleotide (MO) injection in zebrafish eggs resulted in small eye size in 84.38% of the injected larvae, and this phenotype was rescued in 61.39% of the zebrafish eggs when the cpsf1 MO and the cpsf1 mRNA were co-injected. The projection of retinal ganglion cell (RGC) towards the tectum was abnormal in cpsf1 morphants. Thus, we demonstrated that heterozygous LoF mutations in CPSF1 are associated with eoHM and that CPSF1 may play an important role in the development of RGC axon projection.

Updated functional segregation of retinal ganglion cell projections in the tectum of a cyprinid fish-further elaboration based on microelectrode recordings.

Single-unit responses of retinal ganglion cells (GCs) were recorded extracellularly from their axonal terminals in the tectum opticum (TO) of the intact fish (goldfish, carp). The depths of retinal units consecutively recorded along the track of the microelectrode were measured. At the depth of around 50μm, the responses of six types of direction-selective (DS) GCs were regularly recorded. Responses of two types of orientation-selective (OS) GCs and detectors of white and black spots occurred approximately 50μm deeper. Responses of GCs with dark- and light-sustained activity were recorded deeper than all others, at about 200μm. The receptive fields of consecutively recorded units overlap, so they analyze the same fragment of the visual scene, focused by eye optic on the photoreceptor raster. The responses of pairs of DS GCs (ON and OFF units that preferred same direction of stimulus movement) and OS GCs (detectors of vertical and horizontal lines) were often simultaneously recorded at one position of the microelectrode. (The paired recordings of certain units amounted about fourth part of all recordings.) This suggests that their axonal arborizations are located close to each other in the tectal retinorecipient layer. Electrophysiological method, thus, allows to indirectly clarify and make precise the morphology of the retino-tectal connections and to establish a morpho-physiological correspondence.

Ultra-lightweight and wide field of view augmented reality virtual retina display based on optical fiber projector and volume holographic lens

A novel see-through virtual retina display (VRD) system is proposed in this Letter. An optical fiber projector is used as the thin-light-beam source, which is modified from a laser scan projector by separating the laser sources and the scan mechanical structure. A synthetic aperture method is proposed for simple, low-cost fabrication of a volume holographic lens with large numerical aperture. These two key performance-enhanced elements are integrated into a lightweight and ordinary-glasses-like optical see-through VRD system. The proposed VRD system achieves a weight of 30 g and a diagonal field of view of 60°.

Dual-phone illumination-imaging system for high resolution and large field of view multi-modal microscopy.

In this paper we present for the first time a system comprised of two mobile phones, one for illumination and the other for microscopy, as a portable, user-friendly, and cost-effective microscopy platform for a wide range of applications. Versatile and adaptive illumination is made with a Retina display of an Apple mobile phone device. The phone screen is used to project various illumination patterns onto the specimen being imaged, each corresponding to a different illumination mode, such as bright-field, dark-field, point illumination, Rheinberg illumination, and fluorescence microscopy. The second phone (a Nokia phone) is modified to record microscopic images about the sample. This imaging platform provides a high spatial resolution of at least 2 μm, a large field-of-view of 3.6 × 2.7 mm, and a working distance of 0.6 mm. We demonstrate the performance of this platform for the visualization of microorganisms within microfluidic devices to gather qualitative and quantitative information regarding microorganism morphology, dimension, count, and velocity/trajectories in the x-y plane.

The absence of retinal input disrupts the development of cholinergic brainstem projections in the mouse dorsal lateral geniculate nucleus

BackgroundThe dorsal lateral geniculate nucleus (dLGN) of the mouse has become a model system for understanding thalamic circuit assembly. While the development of retinal projections to dLGN has been a topic of extensive inquiry, how and when nonretinal projections innervate this nucleus remains largely unexplored. In this study, we examined the development of a major nonretinal projection to dLGN, the ascending input arising from cholinergic neurons of the brainstem. To visualize these projections, we used a transgenic mouse line that expresses red fluorescent protein exclusively in cholinergic neurons. To assess whether retinal input regulates the timing and pattern of cholinergic innervation of dLGN, we utilized the math5-null (math5−/−) mouse, which lacks retinofugal projections due to a failure of retinal ganglion cell differentiation.ResultsCholinergic brainstem innervation of dLGN began at the end of the first postnatal week, increased steadily with age, and reached an adult-like pattern by the end of the first postnatal month. The absence of retinal input led to a disruption in the trajectory, rate, and pattern of cholinergic innervation of dLGN. Anatomical tracing experiments reveal these disruptions were linked to cholinergic projections from parabigeminal nucleus, which normally traverse and reach dLGN through the optic tract.ConclusionsThe late postnatal arrival of cholinergic projections to dLGN and their regulation by retinal signaling provides additional support for the existence of a conserved developmental plan whereby retinal input regulates the timing and sequencing of nonretinal projections to dLGN.

Measuring the perceptual grouping of non-adjacent surfaces that are invisibly (amodally) or visibly connected.

Classic Gestalt examples of perceptual grouping entail arrays of disconnected surfaces that are grouped on the basis of the surfaces’ relative similarity or proximity. However, most natural environments contain multiple objects, each with multiple, connected surfaces. Moreover, an object in a scene is likely to partially occlude other objects in the 2-dimensional retinal projection of the scene. A central question, therefore, is how the visual system forms a 3-dimensional representation of multi-object scenes by determining which surfaces belong to which objects. To this end, a recently developed dynamic grouping methodology determines whether pairs of surfaces are grouped together on the basis of the direction in which motion is perceived across a surface when its luminance is perturbed. It is shown using this method that the visible surfaces of a partially occluded object are perceptually grouped when they are plausibly connected and represented in a depth plane behind the occluding object. Invisible connectivity (amodal completion) as well as connectivity established by a visible surface have a powerful influence on the grouping of surfaces. However, for neither kind of connectivity is grouping affected by the distance between the surfaces. This absence of a distance/proximity effect on grouping is obtained when the space between to-be-grouped surfaces is filled with other surfaces. It contrasts with the strong effect of distance/proximity on the grouping of disconnected surfaces, and on the clarity of illusory contours formed between disconnected contours. It is concluded that distance/proximity is an operative grouping variable only when there is empty space between the to-be-grouped surfaces.

Suprachiasmatic function in a circadian period mutant: Duper alters light-induced activation of vasoactive intestinal peptide cells and PERIOD1 immunostaining.

Mammalian circadian rhythms are entrained by photic stimuli that are relayed by retinal projections to the core of the suprachiasmatic nucleus (SCN). Neuronal activation, as demonstrated by expression of the immediate early gene c-fos, leads to transcription of the core clock gene per1. The duper mutation in hamsters shortens circadian period and amplifies light-induced phase shifts. We performed two experiments to compare the number of c-FOS immunoreactive (ir) and PER1-ir cells, and the intensity of staining, in the SCN of wild-type (WT) and duper hamsters at various intervals after presentation of a 15-min light pulse in the early subjective night. Light-induced c-FOS-ir within 1hr in the dorsocaudal SCN of duper, but not WT hamsters. In cells that express vasoactive intestinal peptide (VIP), which plays a critical role in synchronization of SCN cellular oscillators, light-induced c-FOS-ir was greater in duper than WT hamsters. After the light pulse, PER1-ir cells were found in more medial portions of the SCN than FOS-ir, and appeared with a longer latency and over a longer time course, in VIP cells of duper than wild-type hamsters. Our results indicate that the duper allele alters SCN function in ways that may contribute to changes in free running period and phase resetting.

Growth Cone Tctp Is Dynamically Regulated by Guidance Cues.

Translationally controlled tumor protein (Tctp) contributes to retinal circuitry formation by promoting axon growth and guidance, but it remains unknown to what extent axonal Tctp specifically influences axon development programs. Various genome-wide profiling studies have ranked tctp transcripts among the most enriched in the axonal compartment of distinct neuronal populations, including embryonic retinal ganglion cells (RGCs), suggesting its expression can be regulated locally and that this may be important during development. Here, we report that growth cone Tctp levels change rapidly in response to Netrin-1 and Ephrin-A1, two guidance cues encountered by navigating RGC growth cones. This regulation is opposite in effect, as we observed protein synthesis- and mTORC1-dependent increases in growth cone Tctp levels after acute treatment with Netrin-1, but a decline upon exposure to Ephrin-A1, an inhibitor of mTORC1. Live imaging with translation reporters further showed that Netrin-1-induced synthesis of Tctp in growth cones is driven by a short 3′untranslated region (3′UTR) tctp mRNA isoform. However, acute inhibition of de novo Tctp synthesis in axons did not perturb the advance of retinal projections through the optic tract in vivo, indicating that locally produced Tctp is not necessary for normal axon growth and guidance.

Light Prior to Eye Opening Promotes Retinal Waves and Eye-Specific Segregation

Retinal waves are bursts of correlated activity that occur prior to eye opening and provide a critical source of activity that drives the refinement of retinofugal projections. Retinal waves are thought to be initiated spontaneously with their spatiotemporal features dictated by immature neural circuits. Here we demonstrate that, during the second postnatal week in mice, changes in light intensity dictate where and when a subset of retinal waves are triggered via activation of conventional photoreceptors. Propagation properties of triggered waves are indistinguishable from spontaneous waves, indicating that theyare activating the same retinal circuits. Using whole-brain imaging techniques, we demonstrate that light deprivation prior to eye opening diminishes eye-specific segregation of the retinal projections to the dorsolateral geniculate nucleus of the thalamus, but not other retinal targets. These data indicate that light that passes through the closed eyelids plays a critical role in the development of the image-forming visual system.

The Pax2 and Pax6 Transcription Factors in the Optic Nerve and Brain of Trout Oncorhynchus mykiss after a Mechanical Eye Injury

The distribution of the Pax2+ transcription factor in the optic nerve after a mechanical eye injury on the side of damage and in the contralateral nerve has been studied in the trout Oncorhynchus mykiss. It has been found that injury of the optic nerve in this fish species causes Pax2+ reactive astrocytes involved in the initial stages of optic nerve axon regeneration to increase in number, especially in the area of the head and the proximal part of the optic nerve. As the optic nerve in trout is damaged, a significant growth of the heterogeneous population of Pax6+ cells occurs in the brain divisions that have direct retinal inputs, diencephalon, and optic tectum. A part of the Pax6+ cells have an undifferentiated phenotype and are a component of reactive neurogenic niches located in the periventricular zone and parenchymal regions of the brain. Another population of Pax6+ cells has the radial glial phenotype and appears as a result of activation of the constitutive neurogenic domains also within the newly formed reactive neurogenic niches. Thus, due to the optic nerve injury, a pronounced neurogenic response associated with the appearance of reactive neurogenic niches and radial glia arises both in the brain divisions with direct retinal projections and in those lacking the retinal projections as well as in remote regions. The results obtained indicate that the damage to the optic nerve causes an increased reactive neurogenesis in the brain of adult trout.

Nodes of Ranvier in Glaucoma

Retinal ganglion cell axons of the DBA/2J mouse model of glaucoma, a model characterized by extensive neuroinflammation, preserve synaptic contacts with their subcortical targets for a time after onset of anterograde axonal transport deficits, axon terminal hypertrophy, and cytoskeletal alterations. Though retrograde axonal transport is still evident in these axons, it is unknown if they retain their ability to transmit visual information to the brain. Using a combination of in vivo multiunit electrophysiology, neuronal tract tracing, multichannel immunofluorescence, and transmission electron microscopy, we report that eye–brain signaling deficits precede transport loss and axonal degeneration in the DBA/2J retinal projection. These deficits are accompanied by node of Ranvier pathology – consisting of increased node length and redistribution of the voltage-gated sodium channel Nav1.6 that parallel changes seen early in multiple sclerosis (MS) axonopathy. Further, with age, axon caliber and neurofilament density increase without corresponding changes in myelin thickness. In contrast to these findings in DBA/2J mice, node pathologies were not observed in the induced microbead occlusion model of glaucoma – a model that lacks pre-existing inflammation. After one week of systemic treatment with fingolimod, an immunosuppressant therapy for relapsing-remitting MS, DBA/2J mice showed a substantial reduction in node pathology and mild effects on axon morphology. These data suggest that neurophysiological deficits in the DBA/2J may be due to defects in intact axons and targeting node pathology may be a promising intervention for some types of glaucoma.