Endothelin-Mediated Changes in Gene Expression in Isolated Purified Rat Retinal Ganglion Cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Rattus norvegicus
PROVIDER: GSE71021 | GEO | 2015/07/17
SECONDARY ACCESSION(S): PRJNA290111
REPOSITORIES: GEO
Similar Datasets
Project description:Goals of the study: 1. Assess the gene expression profile in rat RGC after experimental IOP elevation to elucidate the molecular mechanisms of RGC death. 2. Identify potentially novel genes and pathways that may contribute to RGC death in glaucoma. Background: Glaucoma is a neurodegenerative disease characterized by the slow, progressive degeneration of RGC. Elevated IOP is the biggest risk factor for developing glaucoma, loss of RGC and optic nerve atrophy. The pathophysiology of glaucomatous neurodegeneration is not fully understood. It is now clear that RGC die by apoptosis in glaucoma. However, what triggers the apoptosis is still unknown. So far, several gene expression studies have been performed on the retina as a whole. These studies revealed up and downregulation of many genes in response to elevated IOP and optic nerve transection. However, the retina is a complex tissue composed of neuronal, glial and vascular cell types. The RGCs only comprise 5% or less of retinal cells. The gene expression profiles from whole retina can not represent of RGC gene expression. In the current study, we sought to investigate the whole genome regulation of the RGCs in glaucoma. The study was carried out in 3 adult male Brown Norway rats with experimental glaucoma. An equal number of RGCs were captured from normal eyes and eyes with elevated IOP. Gene expression in the glaucomatous RGC was compared with that in the fellow RGC by using Affymetrix Gene chip.
2010-10-01 | E-GEOD-12596 | biostudies-arrayexpress
Project description:Retinal ganglion cells (RGCs) serve as the essential connection between the eye and the brain, with this connection disrupted in blinding disorders such as glaucoma. Numerous cellular mechanisms have been associated with glaucomatous neurodegeneration, and useful models for the study of glaucoma allow for the precise analysis of these degenerative phenotypes. Human pluripotent stem cells (hPSCs) serve as powerful tools for the in vitro analysis of human neurodegenerative diseases, particularly those cellular mechanisms underlying degeneration. Thus, efforts of the current study were initially focused upon the use of hPSCs with an E50K mutation in the Optineurin (OPTN) gene underlying glaucomatous neurodegeneration. CRISPR/Cas9 gene editing approaches were used to introduce the OPTN(E50K) mutation into existing lines of hPSCs, as well as the generation of isogenic control lines from existing OPTN(E50K) patient-derived hPSC lines. When grown from glaucomatous sources, hPSC-derived RGCs developed similarly to isogenic controls. Conversely, at later stages of maturation, OPTN(E50K) RGCs exhibited numerous deficits associated with a neurodegenerative phenotype, including neurite retraction, autophagy pathway disruption, apoptosis, and increased excitability. The results of this study provide an extensive analysis of the OPTN(E50K) mutation in hPSC-derived RGCs, with a number of biochemical and molecular alterations associated with functional changes to these RGCs. The opportunity now exists to further explore the precise cellular pathways leading to RGC death in glaucoma, as well as develop novel translational approaches for glaucoma including pharmacological screening and cell replacement therapies.
2020-02-11 | GSE145069 | GEO
Project description:Goals of the study: 1. Assess the gene expression profile in rat RGC after experimental IOP elevation to elucidate the molecular mechanisms of RGC death. 2. Identify potentially novel genes and pathways that may contribute to RGC death in glaucoma. Background: Glaucoma is a neurodegenerative disease characterized by the slow, progressive degeneration of RGC. Elevated IOP is the biggest risk factor for developing glaucoma, loss of RGC and optic nerve atrophy. The pathophysiology of glaucomatous neurodegeneration is not fully understood. It is now clear that RGC die by apoptosis in glaucoma. However, what triggers the apoptosis is still unknown. So far, several gene expression studies have been performed on the retina as a whole. These studies revealed up and downregulation of many genes in response to elevated IOP and optic nerve transection. However, the retina is a complex tissue composed of neuronal, glial and vascular cell types. The RGCs only comprise 5% or less of retinal cells. The gene expression profiles from whole retina can not represent of RGC gene expression. In the current study, we sought to investigate the whole genome regulation of the RGCs in glaucoma.
2010-10-01 | GSE12596 | GEO
Project description:Experimental ocular hypertension (IOP) induces senescence of retinal ganglion cells (RGCs) that mimicks events occurring in human glaucoma. An established transgenic p16-3MR mouse model in which the systemic administration of the small molecule ganciclovir (GCV) selectively kills p16INK4a-expressing cells was used to compare transcriptomes of retinas from IOP and control eyes in GCV-treated and non-treated mice, to investigate how experimental removal of senescent p16INK4a-positive cells impacts retinal cells in conditions resembling glaucoma.
2020-12-01 | GSE141725 | GEO
Project description:Retinal ganglion cells (RGCs) convey the major output of information collected from the eye to the brain. RGCs are irreversibly lost when injured in degenerative diseases such as glaucoma; this failure can be partially reversed by eliminating the retinal mobile zinc (Zn2+) and leads to substantial axon regeneration. ZnT3 conditional knockout in retinal amacrine cells blocks the synaptic transport of Zn2+, which promotes RGC survival and axonal regeneration in optic nerve jinjury (ONC). Here, we conducted an mRNA sequencing of flow cytometry-isolated RGCs 3 days after optic nerve injury with or without ZnT3 expression in retinal amacrine cells.
2023-07-01 | GSE213937 | GEO
Project description:Different sub-types of ganglion cells existing in mammalian retinas possessing distinct function in processing the visual information. In primates, RGC are commonly divided into midget cells, which are relatively small, and parasol cells, with a larger soma size. Rodents also possess morphologically distinct populations, but their physiological properties are less characterized. The two types of mammalian RGCs differ in their response to pathological conditions such as retinal ischemia, diabetic retinopathy and glaucoma. In this work, we compared gene expression profiles of large (LRGCs) and small (SRGCs) ganglion cells isolated from rat retina in attempt to identify molecular determinants underlying differences in function and tolerance to stress. Experiment Overall Design: Rat RGCs retrogradely labeled with 4DI-10ASP were purified from freshly dissected retinas and subjected to fluorescent-activated cell sorting (FACS) in order to separate the SRGC and LRGC sub-populations. Actinomycin D was added to prevent new transcription during experimental procedures. Purified cells were used for RNA isolation. Following two rounds of linear amplification aRNA probes were hybridized with two-color Agilent Rat Genomic Oligo Arrays. We utilized the dye swap experimental design to eliminate the dye bias effects and make data from different experiments available for the cross-comparison.
2008-06-20 | E-GEOD-11468 | biostudies-arrayexpress
Project description:Elevated intraocular pressure (IOP) is the major risk factor for glaucoma, a sight threatening disease of retinal ganglion cells (RGCs) and their axons. Despite the central importance of IOP, details of the impact of IOP elevation on RGC gene expression remain elusive. We developed a novel 4-step immunopanning protocol to extract adult mouse RGCs with high fidelity and used it to isolate RGCs from wild type mice exposed to 2 weeks of IOP elevation generated by the microbead model. IOP was elevated to 2 distinct levels which were defined as Mild (IOP increase >1mmHg and <4mmHg) and Moderate (IOP increase > 4mmHg). RNA sequencing was used to compare the transcriptional environment at each IOP level. Differentially expressed genes were markedly different between the 2 groups, and pathway analysis revealed frequently opposed responses between the IOP levels. These results suggest that the magnitude of IOP elevation has a critical impact on RGC transcriptional changes. Furthermore, it is possible that an IOP-based set point exists within RGCs to impact the direction of transcriptional change. It is possible that this improved understanding of changes in RGC gene expression can ultimately lead to novel diagnostics and therapeutics for glaucoma.
2019-02-22 | GSE122205 | GEO
Project description: Not available
| S-EPMC5102497 | biostudies-literature
Project description:Retinal ganglion cells (RGCs) convey the major output of information collected from the eye to the brain. RGCs are irreversibly lost when injured in degenerative diseases such as glaucoma; this failure can be partially reversed by eliminating the retinal mobile zinc (Zn2+) and leads to substantial axon regeneration. Here, we conducted an mRNA sequencing of flow cytometry-isolated RGCs 3 days after optic nerve injury (NC) with or without TPEN treatment. We identified differentially expressed genes using a cutoff of at least 1.5-fold change and FDR of at least 0.05. We focused on the intersecting genes that were upregulated after NC and recovered in TPEN (TPEN/NC; fold change < -1.5; P < 0.05), namely, TPEN target genes.
2022-11-02 | GSE208777 | GEO
Project description:Since retinal ganglion cells (RGCs) do not regenerate after injury, RGC replacement therapy could provide an approach to vision restoration in glaucoma and other optic neuropathies. Here we developed a rapid protocol for direct induction of RGC (iRGC) differentiation from human iPSCs and ESCs in less than two weeks, by overexpression of NGN2 in RGC survival-promoting medium supplemented with a Notch inhibitor. Neuronal morphology and neurite growth were observed within one week of induction. Immunostaining and qRT-PCR for characteristic RGC-specific genes confirmed identity. Calcium imaging was used to evaluate iRGCs’ electrophysiologic maturation, and demonstrated GABA-induced excitatory calcium influx characteristic of immature RGCs. Using single cell-RNA sequencing (scRNA-seq) we further delineated iRGCs’ differentiation and compared iRGC transcriptomic profiles to fetal human and retinal organoid-derived RGCs. Unbiased clustering showed high similarities of transcriptomic profiles among iRGCs, early-stage fetal human RGCs and early-stage retinal organoid-derived RGCs. However, for some markers, including BRN3a, BRN3b and NEFL, iRGCs demonstrated expression patterns more similar to fetal RGCs than to retinal organoid RGCs. After intravitreal injection into rodent eyes, transplanted iRGCs survived and migrated into host retinas where they were detected one week and one month after transplant. Prior optic nerve trauma to the recipient host did not significantly enhance or detract from iRGC integration, but iRGCs protected host RGCs from neurodegeneration. Taken together, these data demonstrate rapid iRGC generation in vitro into an immature cell with high similarity to human fetal RGCs and capacity for retinal integration after transplant, including after optic nerve injury. The simplicity of this system may benefit translational studies on human RGCs.
2022-04-10 | GSE200452 | GEO