Hibernation in a Dish?: Exploring the Ground Squirrel Induced Pluripotent Stem Cells (iPSCs)
Jingxing Ou1, Barbara Mallon2, Kiyoharu Miyagishima1, Wei Li1
1Retinal Neurophysiology Section, National Eye Institute; 2Stem Cell Unit, National Institute of Neurological Disorders & Stroke, National Institutes of Health
Hibernation is an effective strategy adopted by some animals to survive the harsh winter conditions. During hibernation, thirteen-lined ground squirrels (Ictidomys tridecemlineatus) lower their core body temperature to 2-8 °C. When awakened from deep torpor, they can regain normal body temperature and motility within several hours. However, such drastic changes are stressful to the animal. For example, in many species, the microtubules, a major cytoskeletal component that neurons rely on to transport vital signals and materials and form neural circuits, dissemble and hence sabotage neuronal activity when the temperature drops substantially below 37 °C. To study cellular changes and protective mechanisms under hibernation-like conditions in ground squirrels, a versatile in vitro tool would be very useful. We describe here the first successful generation of ground squirrel induced pluripotent stem cell (iPSC) lines created to address this need. Pluripotency of the cell lines was verified by both in vitro differentiation and in vivo teratoma formation and karyotyping confirmed the lines had the appropriate chromosome number for this species. Primary neonatal cortical cultures of ground squirrel neurons and iPSC-derived ground squirrel neurons were cultured at 4-8 °C. for several days to test their resilience compared to human iPSC derived neurons. We found that both ground squirrel neuronal cultures were indeed resilient to the temperature drop, maintaining their neuronal processes for several days, while the microtubule processes of human iPSC-derived neurons collapsed overnight. Further analyses on ground squirrel and human iPSC-derived neuronal transcriptomes and microtubule dynamics may unveil novel neural protective mechanisms that can inspire effective strategies to treat human neuronal injuries and diseases.
Development of the First Visual Synapse
D. Thad Whitaker1,2, Passley Hargrove1,3, Soo-Young Kim1, Amal Alsufyani1,4, Jung-Woong Kim1, Hyun Jin Yang1, Hannah Fann1, Kristen Mollura1, Anand Swaroop1
1Retinal Development, Genetics & Therapy Section, Neurobiology Neurodegeneration & Repair Laboratory, National Eye Institute, National Institutes of Health; 2Texas A&M Institute for Neuroscience, Texas A&M University; 3Institute of Biomedical Sciences, George Washington University; 4Department of Chemistry & Biochemistry, George Mason University
Neuronal morphology, including accurate targeting of the axon and the appropriate weight of the presynaptic area, are key factors in a neuron’s functionality and performance. Screens to identify genes involved in axon development/regeneration, synapse formation, and axon targeting have been performed using in vitro/ex vivo cultures but much less work has been performed in a native system. We set out to determine transcriptional targets of NRL (the transcription factor which determines the cell fate of rod photoreceptors) that are important for synapse morphology. We injected short hairpin RNAs (shRNAs) into newborn mouse retinas, along with a fluorescent reporter under a photoreceptor specific promoter, by in vivo electroporation. After maturation of the eye, the retinas were removed, sectioned, and stained. Individual pre-synaptic segments were imaged and measured for three criteria (synapse width, relative position within the OPL, and presence/absence of telodendrites) by multiple blind observers. Values were averaged for each synapse then each gene-specific shRNA was compared to the scrambled shRNA and no-shRNA controls. We have found eight potential constructs to date that modulate synapse morphology following gene knockdown in the three criteria measured. These are still to be verified using additional shRNAs and rescue experiments but already give promising leads to future avenues of study. Our results begin to shed light on genes previously unknown to play a role in the formation of neuronal morphology. This work also shows that the retina, specifically photoreceptors, provides a good, native system for study of modulators of synapse size and positioning.
NK-DC Crosstalk Controls the Uveitogenic Th17 Response Through an Innate IFN-γ/IL-27 Axis
Wai Po Chong1, Nicolas van Panhuys2, Jun Chen1, Phyllis Silver1, Ronald Germain2, Rachel Caspi1
1Immunoregulation Section, Laboratory of Immunology, National Eye Institute, 2Lymphocyte Biology Section, Laboratory of Systems Biology, National Institute of Allergy & Infectious Diseases, National Institutes of Health
Autoimmune uveitis is caused by the inflammation in eye tissues that can lead to severe vision handicap. Because of the strong MHC association with the disease, it is believed that the MHC molecules present eye antigen to activate T cells and trigger the inflammation and tissue damage. Upon activation, T cells differentiate into Interferon (IFN)-γ producing T helper (Th)1 cells or Interleukin (IL)-17A producing Th17 cells. IFN-γ is a pathogenic cytokine involved in inflammation. Paradoxically, its deficiency exacerbates experimental autoimmune uveitis. We previously showed that it is innate IFN-γ that is protective, whereas adaptive IFN-γ from T cells is pathogenic. Here we demonstrate by immunological and imaging methods, using IFN-γ-deficient mice repleted with IFN-γ-sufficient Natural Killer (NK) cells, that innate production of IFN-γ from NK cells is necessary and sufficient to limit autoimmunity and to reduce the number of IL-17A and granulocyte macrophage colony-stimulating factor (GM-CSF) producing effector T cells in the eye. Mechanistic studies demonstrated that, following immunization for uveitis, dendritic cell (DC) recruited IFN-γ-producing NK cells to the draining lymph node (DLN) and interacted with them in a CXCR3-dependent fashion. During the interaction, IFN-γ from NK cells induced DCs to produce IL-27, which in turn enhanced IFN-γ production by NK cells, forming a positive feedback loop. The NK-DC-dependent IL-27 inhibited development of the adaptive pathogenic IL-17 response. Our data reveal that an early NK-DC interaction controls the adaptive Th17 response and limits tissue-specific autoimmunity through an innate IFN-γ/IL-27 axis.
Sensorimotor Encoding in Basal Ganglia During a Covert Attention Task
Fabrice Arcizet, Richard Krauzlis
Section on Eye Movements & Visual Selection, Laboratory of Sensorimotor Research, National Eye Institute, National Institutes of Health
Attention plays a crucial role in seeing what is important and choosing the correct action. Disorders of attention are also fairly common (e.g., ADHD). The complete brain circuit for attention is not yet known, but one possibility is a pathway through the basal ganglia. Here we investigated how neuronal activity in the caudate nucleus, a major input structure of the basal ganglia, is modulated by different task events during the performance of a visual attention task.
To study attention, we trained macaque monkeys to perform a motion-change detection task. The task was to release a joystick when one of two motion patches changed its direction (“the cued” patch) and to ignore changes at the other location (the “foil”). Animals performed the task well, correctly reporting “cued” changes on 62% of trials and incorrectly reporting “foil” changes on only 5%.
We found that caudate neurons were modulated by several factors during the attention task. Early during the trial, after the appearance of the cue and motion patches, the most significant factor for a plurality of caudate cells (30%) was cue location. Later in the trial, when the motion change could happen, caudate cells were more strongly modulated by the response choice (release or not) of the monkey (37%). Importantly, this response choice activity depended on both sensory and motor factors – caudate neurons showed significant preferences for particular stimulus locations and also predicted whether or not the monkey would release the joystick.
These results illustrate how the caudate nucleus may contribute to the performance of visual attention tasks – by linking sensory selectivity to particular motor actions, caudate activity could be used to select the action appropriate for the sensory context. By identifying a novel component of the brain circuitry for attention, these results are a step toward identifying the etiology of attention disorders.
Developmentally Guided Protocol for Efficient Differentiation of Induced Pluripotent Stem Cells to Retinal Pigment Epithelium
Ruchi Sharma1, Janine Davis1, Vladimir Khristov2, Shekhar Jha1, Qin Wan2, Fang Hua2, Kapil Bharti1
1Unit on Ocular & Stem Cell Translational Research, 2Section on Epithelial & Retinal Physiology & Disease, Ophthalmic Genetics & Visual Function Branch, National Eye Institute, National Institutes of Health
Induced pluripotent stem (iPS) cells have the potential to differentiate into any somatic cell type and hence can be used to study disease mechanism and develop cell-based therapies. Retinal degeneration a blinding eye disease that can occur because of Retinal Pigment Epithelium (RPE) cell dysfunction and atrophy. RPE helps in maintenance of photoreceptor health and function. Age Related Macular Degeneration (AMD) is one of the retinal degenerations, a foremost cause for blindness in developed countries. Currently there is no cure for “dry” form of AMD where RPE cells die-off. A cell-based therapy using iPS cell derived RPE could be used to treat this blinding disease. Here we have developed a differentiation protocol for the derivation of RPE monolayer from iPS cells. We used a dual reporter iPS cell line expressing RPE specific GFP and constitutive RFP to optimize the differentiation protocol. The triphasic developmentally guided protocol marks the commitment of iPS cells towards RPE-primed neuroectoderm using a dual SMAD, canonical WNT, and FGF inhibition. Committed RPE cell differentiation requires an activation of Canonical WNT and TGF-signaling pathways and maturation of RPE cells requires a down regulation of canonical WNT pathway. The first phase of the protocol significantly increases the expression of eye-field transcription factors PAX6, RAX, OTX1, and SIX3; GFP expression significantly increases by using a combination of dual-SMAD and FGF inhibition. This differentiation protocol reproducibly generates RPE at a high efficiency from different healthy and patient-derived iPS cell lines. RPE monolayers derived using this protocol perform RPE functions that resemble native human RPE.
Modulation of Immune Responses by Extracellular Vesicles from Retinal Pigment Epithelium
Jared Knickelbein1, Baoying Liu1, Anush Arakelyan2, Sonia Zicari2, Susan Hannes1, Ping Chen1, Zhiyu Li1, Jean-Charles Grivel2, H. Nida Sen1, Leonid Margolis2, Robert Nussenblatt1
1Clinical Immunology Section, Laboratory of Immunology, National Eye Institute, 2Program in Physical Biology, Eunice Kennedy-Shriver National Institute of Child Health & Human Development, National Institutes of Health
Introduction: Extracellular vesicles (EV), such as exosomes, are important mediators of intercellular communication and have been implicated in modulation of the immune system. We investigated if EV released from retinal pigment epithelium (RPE) modulates immune responses in vitro.
Methods: EV were isolated from ARPE-19 cultures stimulated or not with the inflammatory cytokines IL-1β, IFN-γ, and TNF-α. Isolated EV were characterized with a nanoparticle analyzer and cultured with human peripheral blood mononuclear cells, which were assayed for T cell proliferation by 3H-thymidine incorporation. RPE-derived EV were also independently cultured with enriched lymphocytes or monocytes. Cell phenotype and cell death were evaluated by flow cytometric analysis. Cytokine levels were assayed in culture supernatants by multiplex bead analysis.
Results: The size and concentration of EV secreted from ARPE-19 stimulated or not with inflammatory cytokines did not significantly differ. EV from both resting and cytokine-stimulated ARPE-19 significantly inhibited T cell proliferation without affecting T cell viability. Culture of EV from resting ARPE-19 with undifferentiated human monocytes induced an immunoregulatory phenotype with a significantly higher percentage of CD14++CD16+ monocytes and upregulation of TGF-β1. Culture of EV from cytokine-stimulated ARPE-19 cells with human monocytes induced upregulation of several pro-inflammatory cytokines and monocyte death.
Conclusions: RPE cells constitutively secrete EV in the size range of exosomes. EV from both resting and cytokine-stimulated RPE inhibited T cell stimulation. EV from resting ARPE-19 cells promoted an immunoregulatory CD14++CD16+ phenotype in human monocytes, while EV from cytokine-stimulated ARPE-19 cells caused human monocyte death. These findings suggest that RPE cells use EV to induce a down-regulatory immune environment under homeostatic conditions, and in an inflammatory milieu, RPE-derived EV may mitigate a potentially harmful inflammatory response through killing of immune cells.
Platelet-derived Growth Factor Receptor Alpha (PDGFRα)-mediated Retinal Ganglion Cell Neuroprotection
Shokichi Takahama1, Modupe Adetunji1, Tantai Zhao2, Shan Chen2, Rafael Villasmil3, Wei Li2, Stanislav Tomarev1
1Section on Retinal Ganglion Cell Biology, Laboratory of Retinal Cell & Molecular Biology, 2Retinal Neurophysiology Section, 3Flow Cytometry Core, National Eye Institute, National Institutes of Health
Glaucoma, a group of chronic, degenerative optic neuropathies, is a major cause of blindness worldwide. Retinal ganglion cell (RGC) death is one of the pathogenic features of glaucoma. Our previous study showed that treatment of the retina with platelet-derived growth factor AA (PDGF-AA) rescued RGCs from cell death in a rat glaucoma model and retinal explant culture. PDGF-AA acts through interaction with PDGF receptor alpha (PDGFRα). However, physiological roles of PDGFRα in adult retina and the mechanisms of PDGFRα-mediated neuroprotection are poorly understood. To elucidate the molecular mechanism of PDGFRα-mediated neuroprotection, we utilized an RGC primary culture system and PDGFRα-EGFP mice, in which nuclear-targeted EGFP is expressed under the control of PDGFRα promoter. Using survival assays and immunofluorescent marker studies, we found that PDGF-mediated neuroprotection of RGCs does not occur through a direct interaction of PDGF-AA with RGCs and that RGCs do not express PDGFRα. We demonstrated that astrocytes in the RGC layer and a sub-population of amacrine cells in the inner nuclear layer express PDGFRα. These data suggest that the neuroprotective effect of PDGFRα is mediated by astrocytes and/or a sub-population of amacrine cells that secrete factors protecting RGCs. To identify the neuroprotective factors induced by PDGFRα activation, we examine the gene expression patterns of PDGFRα-expressing GFP positive cells in the healthy retina and in the retina following optic nerve crush isolating these cells by FACS with subsequent RNA sequencing analysis.
PDZPH1, a Newly Identified Protein in Photoreceptor Function and Disease
Vetrivel Sengottuvel1, Lijin Dong2, Tiansen Li1
1Retinal Cell Biology & Degeneration Section, Neurobiology-Neurodegeneration & Repair Laboratory, 2Genetic Engineering Core, National Eye Institute, National Institutes of Health
Intracellular trafficking to photoreceptor outer segment and to the synaptic terminal is a polarized process that involves transport of proteins synthesized and processed in the biosynthetic inner segment. Impairment of protein trafficking linked to mutations in the genes coding for Tubby and Whirlin causes early and late onset retinal degeneration, respectively. We report here the identification of a novel protein, designated PDZPH1, as an interacting partner for both tubby and whirlin. PDZPH1 was discovered in yeast two hybrid screens by virtue of its interaction with tubby and whirlin. It is highly expressed in photoreceptors, conserved across vertebrate species, and restricted to photoreceptors in its expression pattern. PDZPH1 is enriched in tubulo-vesicular structures in photoreceptor apical inner segments. In addition, PDZPH1 also interacts with SNAP25 as shown by co-immunoprecipitation of mouse retinal extracts. CRISPR/Cas9 mediated disruption of PDZPH1 in mice leads to early and pronounced functional deficits in photoreceptors and a late onset degeneration. At an early age (1.5 months), the amplitudes of electroretinograms (ERG) originating from both rods and cones are significantly reduced in the mutant. Notably, the early ERG defects are marked by a substantial decline in b-wave but relative sparing of the a-wave, suggesting a greater involvement of the synaptic terminals. Thinning of the outer plexiform (synaptic) layer and diminished labeling of synaptic proteins are seen at this age. Moreover, aberrant accumulation of early endosomes (marked by EEA1, Rab5) and vacuolar structures in the photoreceptor inner segment are found. Visual deficits and photoreceptor degeneration are slowly progressive such that by 8 months of age, almost 50% of photoreceptors are lost and ERG is nearly flat. We conclude that PDZPH1 functions in endosomal / vesicular cargo transport in photoreceptor cells, with a critical and non-redundant role in directing traffic to the synaptic terminals.
Binocular Correlation Computations in Primary Visual Cortex
Sindre Henriksen1,2, Jenny Read2, Bruce Cumming1
1Vision Section, Laboratory of Sensorimotor Research, National Eye Institute, National Institutes of Health; 2Institute of Neuroscience, Newcastle University, United Kingdom
Binocular (3D) depth perception relies on small differences in the images seen by the left and right eyes, owing to their different placements on the head. In order to extract depth from the difference between the two eyes (retinal disparity), the brain has to solve the stereo correspondence problem: which points in the left eye corresponds to those in the right? The first step in the solution to this problem is generally thought to be carried out by neurons in V1 who perform a correlation-based computation on the images seen by the left and right eyes. Recently, a series of publications have shown that humans can see depth in stimuli with a mean binocular correlation of 0, suggesting that correlation-based V1 cells cannot signal disparity in this case. We have shown that a simple modification to a correlation-based model of V1 cells can produce neurons that are disparity-tuned to zero mean binocular correlation stimuli by exploiting fluctuations in the correlation level. These cells can be combined to produce a psychophysical decision model that performs very similarly to humans and makes predictions about how humans should behave in response to novel stimuli. Lastly, we record from disparity-selective cells in V1 and show that V1 neurons are indeed disparity-tuned to these stimuli, and (like our model cells) appear to be sensitive to the fluctuations in the correlation level.
RetSeq: A Database Integrating Retina-specific High-throughput Data
Koray Dogan Kaya, Vijender Chaitankar, Gökhan Karakulah, Anand Swaroop
Retinal Development, Genetics & Therapy Section, Neurobiology Neurodegeneration & Repair Laboratory, National Eye Institute, National Institutes of Health
The advent of technologies measuring different genomic features in a tissue has led us new ways to understand better the details of how changes in genome affect phenotype. Many of those technologies use Next Generation Sequencing (NGS). Attempts to understand effects of individual features on different tissues under different conditions rarely went further from what we have known, rather they repeated the results of earlier experiments or revealed the well predicted/speculated phenomena. However, we anticipate that we can do better with those technologies providing measurements that are more precise. Instead of trying to mine individual data to address specific research questions, integrating and then mining them would be better. Even if the integration can be tissue specific, we can have more precise results with easier mining.
Here, we have developed a platform with web interface integrating NGS data measuring the retinal (with its sub-cell types) transcriptome and the features of their regulatory context. Our platform reveal very important unknowns related to a wide range of biological questions such as how mouse retina develops, how genes behave during development and diseases and detailed transcriptional mechanisms of mouse retinal disease models. We are extending our platform for the data coming from other model organisms as well as human.
In vivo Rod Photoreceptor Reprogramming by AAV-delivered CRISPR/Cas9 Rescues Retinal Degeneration
Wenhan Yu1, Suddhasil Mookherjee1, Jung-Woong Kim2, Suja Hiriyanna1, Lijin Dong3, Tiansen Li4, Anand Swaroop2, Zhijian Wu1
1Ocular Gene Therapy Core, 2Retinal Development, Genetics & Therapy Section, Neurobiology Neurodegeneration & Repair Laboratory, 3Genetic Engineering Core, 4Retinal Cell Biology & Degeneration Section, Neurobiology Neurodegeneration & Repair Laboratory, National Eye Institute, National Institutes of Health
Retinitis pigmentosa (RP) is the most common form of inherited retinal dystrophy, characterized by early rod photoreceptor death and secondary cone degeneration. In a recent proof-of-concept study, knocking down the rod photoreceptor determinant Nrl rendered the rods resistant to the effects of mutations in rod-specific genes and consequently prevented secondary cone death. To develop this strategy into a generic treatment for RP, we combined the advantages of CRISPR/Cas9 and adeno-associated viral (AAV) vector for efficient gene knockdown in photoreceptors. Two AAV vectors were included in each gene-specific AAV-CRISPR, one containing a Cas9 expression cassette and another containing a single-guided (sg) RNA targeting the gene locus. After validating the potency of this system by knocking down EGFP in a mouse line with EGFP-labeled rods, we evaluated the Nrl gene disruption in wild-type mice following administration of the Nrl-specific vectors. Insertions/deletions in the targeted Nrl locus were detected in roughly 70% of AAV-transduced photoreceptors by deep DNA sequencing, which caused Nrl reading frame shifts. Consequently, down-regulation of some rod-specific genes and up-regulation of cone-specific genes in transduced rods were detected by RNA-Seq and protein analyses, indicating reprogramming of rods into cone-like cells. To assess the effectiveness of the approach for rescue of retinal degeneration, three mouse models harboring either recessive or dominant rod-specific mutations received vector treatment. In all three models, a majority of the AAV-CRISPR-transduced rods were reprogrammed into cone-like cells and maintained their viability, though the rod function was lost. Cone function and viability in the treated eyes were maintained throughout the duration of the study, as revealed by the preserved cone ERG amplitudes and cone marker staining. In conclusion, AAV-CRISPR-mediated Nrl knockdown can efficiently reprogram rods into cone-like photoreceptors and prevent secondary cone death in retinal degeneration, which could be developed into a viable treatment for RP in humans.
Understanding Genetic Disease through Homology Modeling and Computational Analysis of Human Tyrosinase
Katie Farney, Monika Dolinska, Yuri Sergeev
Ophthalmic Genetics & Visual Function Branch, National Eye Institute, National Institutes of Health
Oculocutaneous albinism type 1 (OCA1) is a genetic disease characterized by the absence of pigment in the skin, hair, and eyes. The mutated protein responsible for this incurable disease is human tyrosinase (Tyr), a type 3 copper glycoprotein that functions as the rate-limiting enzyme controlling the production of melanin pigment in melanosomes. This disease is often diagnosed as OCA1A or OCA1B, where OCA1A is characterized by a complete loss of Tyr enzymatic function and OCA1B has reduced Tyr enzymatic activity. Many deleterious, long-term effects of both OCA1A and OCA1B have been documented, but much remains unidentified about the link between molecular genetics, protein structure, and variations in phenotype.
The crystal structure of human tyrosinase is unknown. However, we have used homology modeling and molecular dynamics to construct a reliable, atomic protein structure that has allowed us to better understand its role in melanin production. Our fully glycosylated, intra-melanosomal domain of human tyrosinase (residues 106 – 451) was used to analyze the Gibbs free energy changes (ΔΔG) between the wild-type and five clinical mutations: T373K, P406L, R402Q, R422Q, and R422W. The computational free energy changes were then compared to our experimental ΔΔG values and a strong correlation was found (Pearson’s R = 0.9207). Our model will allow us to further use computational methods to bridge the knowledge gap between molecular genotype and phenotype, in addition to focusing on Tyr’s enzymatic active site for evaluation of compounds as potential activators of mutant Tyr in OCA1 disorders.
- Molecular and Cytogenetic Evaluations of Atypical Anterior Segment Dysgenesis Syndromes with Cardiac Involvement
Robert Hufnagel1,2, Michael Yang2, Michael Gray2, Laura Krueger2, Thomas Jaworek4, Patricia Bender3, Pei-Wen Chiang5, Xia Li3, Teresa Smolarek3, Howard Saal3, Robert Hopkin2, Zubair Ahmed4
1Ophthalmic Genetics & Visual Function Branch, National Eye Institute, National Institutes of Health; 2Division of Pediatric Ophthalmology; 3Division of Human Genetics, Cincinnati Children’s Hospital Medical Center; 4Department of Otorhinolaryngology-Head & Neck Surgery, University of Maryland School of Medicine; 4Casey Eye Institute Molecular Diagnostics Laboratory, Oregon Health & Science University
- PEDF Alterations that Modify Receptor Affinity: Implications for the construction of PEDF with optimal biological effects
Jeanee Bullock1,2, Federica Polato1, Valeria Marigo3, S. Patricia Becerra1
1Section on Protein Structure & Function, Laboratory of Retinal Cell & Molecular Biology, National Eye Institute, National Institutes of Health; 2Department of Biochemistry & Molecular & Cellular Biology, Georgetown University Medical Center; 3Department of Life Sciences, University of Modena & Reggio Emilia
- High-throughput Screening for Identifying Candidate Drugs to Prevent Proteotoxic Stress-induced Retinal Pigment Epithelium (RPE) Cell Death
Justin Chang1, Balendu Shekhar Jha1, Madhu Lal2, Ruchi Sharma1, Marc Ferrer2, Kapil Bharti1
1Unit on Ocular & Stem Cell Translational Research, Ophthalmic Genetics & Visual Function Branch, National Eye Institute, 2Division of Pre-Clinical Innovation, National Center for Advancing Translational Sciences, National Institute of Health
- Optogenetic Stimulation of Striatum during an Active Visual Detection Task in Mice
Lupeng Wang, Krsna Rangarajan, Richard Krauzlis
Section on Eye Movements & Visual Selection, Laboratory of Sensorimotor Research, National Eye Institute, National Institutes of Health
- Retina-specific T Cells Provide Neuroprotection in a Mouse Model of Glaucoma
Jennifer Kielczewski, Reiko Horai, Rachel Caspi
Immunoregulation Section, Laboratory of Immunology, National Eye Institute, National Institutes of Health
- Human Induced Pluripotent Stem Cell (iPSC) Disease Model of Late-Onset Retinal Degeneration
Zoya Qureshy1, Kiyoharu Miyagishima2, Congxiao Zhang2, Ruchi Sharma1, Katharina Clore-Gronenborn1, Vaisakh Rajan1, Jason Silver1, Qin Wan2, Catherine Cukras3, Paul Sieving4, Sheldon Miller2, Kapil Bharti1
1Unit on Ocular Stem Cell & Translational Research, 2Section on Epithelial & Retinal Physiology & Disease, Ophthalmic Genetics & Visual Function Branch, 3Clinical Trials Branch, Division of Epidemiology & Clinical Applications, 4Office of the Director, National Eye Institute, National Institutes of Health
- Disparity Selective Adaptation to Correlated and Anticorrelated Patterns in V1 Neurons Require Computation Across a Broad Population of Neurons
Paul Aparicio, Bruce Cumming
Vision Section, Laboratory of Sensorimotor Research, National Eye Institute, National Institutes of Health
- Unfolding Patterns of Visual Cycle Proteins
Caitlyn McCafferty, Yuri Sergeev
Ophthalmic Genetics & Visual Function Branch, National Eye Institute, National Institutes of Health
- Spontaneous Ocular Autoimmunity in the Absence of Th1 and Th17 Effector Cytokines
So Jin Bing1,2, Reiko Horai1, Wai Po Chong1, Phyllis Silver1, Yingyos Jittayasothorn1, Chi-Chao Chan3, Rachel Caspi1
1Immunoregulation Section, Laboratory of Immunology, National Eye Institute, National Institutes of Health; 2Department of Veterinary Medicine, Jeju National University, South Korea; 3Immunopathology Section, Laboratory of Immunology, National Eye Institute, National Institutes of Health
- Ciliary Defects of rd16 iPSC-derived Photoreceptors in in vitro Three-dimensional Differentiation
Holly Yu Chen1, Pinghu Liu2, Lijin Dong2, Anand Swaroop1
1Retinal Development, Genetics, & Therapy Section, Neurobiology Neurodegeneration & Repair Laboratory, 2Genetic Engineering Core, National Eye Institute, National Institutes of Health
- Expression, Stability, and Structure of Chicken gS-crystallin
Vatsala Sagar1, Yingwei Chen1, Hoay-Shuen Len1, Jianguo Fan1, Katherine Peterson1, Phillip Wilmarth2, Larry David2, Graeme Wistow1
1Section on Molecular Structure & Functional Genomics, National Eye Institute, National Institutes of Health; 2Proteomics Shared Resource, Oregon Health & Science University
- Probing RPE65 Palmitoylation by Acyl-exchange Labeling
Tingting Liu, Eugenia Poliakov, Susan Gentleman, T. Michael Redmond
Molecular Mechanisms Section, Laboratory of Retinal Cell & Molecular Biology, National Eye Institute, National Institutes of Health
- Ocular Microflora Modulates IL-17 and Neutrophil Recruitment within Conjunctiva Associated Lymphoid Tissue
Anthony St. Leger1, Rebecca Drummond2, Desai Jigar2, Phyllis Silver1, Mihalis Lionakis2, Rachel Caspi1
1Immunoregulation Section, Laboratory of Immunology, National Eye Institute; 2Fungal Pathogenesis Unit, Laboratory of Clinical Infectious Diseases, National Institute of Allergies & Infectious Diseases, National Institutes of Health
- Identification of Rare Variants Associated with Age-related Macular Degeneration by Whole Exome Sequencing in 41 Extended Families
Rinki Ratnapriya1, Maartje Geerlings2, Eiko De Jong2, Jordi Corominas Galbany2, Lars Fritsche3, Kari Branham4, Nicole Saksens2, Samuel Perez1, Vijender Chaitankar1, Mohammad Othman4, Deborah Ferrington5, Emily Chew6, Goncalo R. Abecasis3, Carel Hoyng2, Anneke den Hollander2, Anand Swaroop1
1Retinal Development, Genetics, & Therapy Section, Neurobiology Neurodegeneration & Repair Laboratory, National Eye Institute, National Institutes of Health; 2Department of Ophthalmology, Radboud University Medical Center, Nijmegen, The Netherlands; 3Department of Biostatistics, 4Department of Ophthalmology, University of Michigan; 5Departments of Ophthalmology & Neurosciences, University of Minnesota Twin Cities; 6Clinical Trials Branch, Division of Epidemiology & Clinical Applications, National Eye Institute, National Institutes of Health
- Modeling Synaptic Vesicle Dynamics in Active vs. Hibernating Ground Squirrel Cones Predicts Hidden Ribbon Functions
John Ball1, Shan Chen1, Cole Graydon2, Nicholas Brecha3, Bechara Kachar2, Wei Li1
1Retinal Neurobiology Section, National Eye Institute, 2Section on Structural Cell Biology, Laboratory of Cell Structure & Dynamics, National Institute on Deafness & Communication Disorders, National Institutes of Health; 3Department of Neurobiology, University of California, Los Angeles
- Atypical Cadherin FAT1 is Essential for Optic Fissure Closure in Mouse and Zebrafish
Aman George1, Sanita Bharti1, Ruchi Sharma2, Felix Onojafe1, Jerry Lee1, Sunit Dutta1, Helen McNeill3, Kapil Bharti2, Brian Brooks1
1Pediatric, Developmental & Genetic Ophthalmology Section, 2Section on Epithelial & Retinal Physiology & Disease, Ophthalmic Genetics & Visual Function Branch, National Eye Institute, National Institutes of Health; 3Lunenfeld- Tanenbaum Research Institute, Mt. Sinai Hospital, & Department of Molecular Genetics, University of Toronto