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Large-scale screen of novel zebrafish retinal ganglion cell ablation model reveals genetic regulation of retinal regeneration is context specific

ABSTRACT: Large-scale screen of novel zebrafish retinal ganglion cell ablation model reveals genetic regulation of retinal regeneration is context specific

PROVIDER: PRJNA1115053 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR29141400_1.fastq.gz	Fastqsanger.gz
	SRR29141400_2.fastq.gz	Fastqsanger.gz
	SRR29141500_1.fastq.gz	Fastqsanger.gz
	SRR29141500_2.fastq.gz	Fastqsanger.gz
	SRR29141401_1.fastq.gz	Fastqsanger.gz

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Large-scale screen of novel zebrafish retinal ganglion cell ablation model reveals genetic regulation of retinal regeneration is context specific

Project description:Many genes are known to regulate Müller glia (MG)-dependent retinal regeneration following widespread tissue damage. Conversely, genes controlling regeneration following limited retinal cell loss, per degenerative disease, are undefined. Studying regeneration in the context of selective cell loss is important as evidence suggests injury specifics inform the regenerative process. Here, transgenic zebrafish enabling inducible selective retinal ganglion cell (RGC) ablation were combined with single cell multiomics and CRISPR/Cas9-based knockout methods to screen 101 genes for effects on RGC regeneration. We identified 18 regulators of RGC regeneration- seven knockouts inhibited and eleven promoted RGC regeneration. Surprisingly, 35 of 36 known/implicated regulators of retinal tissue regeneration following widespread damage were not required for RGC regeneration, and seven of these knockouts actually enhanced RGC replacement kinetics, including sox2, olig2, and ascl1a. Mechanistic analyses revealed ascl1a knockout increased the propensity of progenitor cells to produce RGCs; i.e., biased progenitor cell fate. These data demonstrate plasticity in how MG can convert to a stem-like state and context-specificity in how genes function during regeneration. Increased understanding of how disease-relevant cell types can be selectively regenerated will, support the development of disease-tailored regenerative therapeutics.

2024-05-27 | GSE268179 | GEO

Molecular Profiling of Small and Large Retinal Ganglion Cells

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Transcription profiling of rat small and large retinal ganglion cells

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

RNA-seq analysis of RNA from DBA/2J retinal ganglion cells

Project description:RNA-seq analysis from young and pre-glaucomatous DBA/2J retinal ganglion cells and control (age and sex-matched, D2-Gpnmb+) retinal ganglion cells

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Subcellular expression profiling of the growth cones of retinal ganglion cells (RGC)

Project description:Cue-directed axon guidance depends partly on local translation in growth cones. Many mRNA transcripts are known to reside in developing axons yet little is known about their subcellular distribution or, specifically, which transcripts are in growth cones. Laser capture microdissection (LCM) was used to isolate the growth cones of retinal ganglion cell (RGC) axons of two vertebrate species, mouse and Xenopus, coupled with unbiased genome-wide microarray profiling. Localized mRNA from the isolated growth cones of Xenopus laevis and Mus musculus retinal ganglion cells were subjected to microarray analysis

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Transcription profiling of mouse developing retinal ganglion and amacrine cells reveals molecular heterogeneity of

Project description:During development of the central nervous system (CNS), cycling uncommitted progenitor cells give rise to a variety of distinct neuronal and glial cell types. As these different cell types are born, they progress from newly specified cells to fully differentiated neurons and glia. In order to define the developmental processes of individual cell types, single cell expression profiling was carried out on developing ganglion and amacrine cells of the murine retina. Individual cells from multiple developmental stages were isolated and profiled on Affymetrix oligonucleotide arrays. These experiments have yielded an expanded view of the processes underway in developing retinal ganglion and amacrine cells, as well as several hundred new marker genes for these cell types. In addition, this study has allowed for the definition of some of the molecular heterogeneity both between developing ganglion and amacrine cells and among subclasses of each cell type. Experiment Overall Design: Single retinal cells were isolated in tubes containing lysis buffer, their mRNAs were reverse transcribed, and the resulting cDNAs were PCR amplified for 35 cycles. Labeled cDNA samples were hybridized to Affymetrix 430 2.0 microarrays and the data was normalized using MAS5.0 software. Cells were identified post hoc as either developing retinal ganglion cells, amacrine cells or rod photoreceptor cells.

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2025-07-23 | GSE303399 | GEO

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Project description:Retinal ganglion cells (RGCs) are the projection neurons in the retina that connect the visual sensing tissue to the brain. We found that Ascl1/Brn3b/Isl1 transcription factor combination can quickly and efficiently reprogramming mouse embryonic fibroblasts (MEFs) into retinal ganglion cell-like neurons (iRGCs). Using RNA-seq, we analyzed the transcriptomes of MEFs infected with Ascl1/Brn3b/Isl1-overexpressing viruses on day 2 or day 7 of reprogramming, or the final iRGCs on day 13 of reprogramming.

2020-09-30 | GSE140689 | GEO

Direct induction of retinal ganglion cell-like neurons

Project description:Mouse embryonic fibroblasts were reprogrammed into retinal ganglion cell-like neurons by overexpressing Ascl1/Brn3b/Isl1 transcription factor combination in 13 days.

2020-09-30 | GSE140128 | GEO

RNA-Seq and ChIP-Seq gene profiling of retinal ganglion cells in Math5Cre-driven Knockout of Ezh2 and G9a in murine retinal ganglion cells at P0

Project description:Epigenetic mark deposition during embryonic development contribute to postnatal homeostasis and tissue stability. Previously, we found out that Ezh2 contributes critically to the function and postnatal cell survival in bipolar cells but not in retinal ganglion cells in the retina. (Yan et al. Postnatal onset of retinal degeneration by loss of embryonic Ezh2 repression of Six1. Sci Report. doi:10.1038/srep33887; (Cheng L, Wong LJ, Yan N, Han RC et al. Ezh2 does not mediate retinal ganglion cell homeostasis or their susceptibility to injury. PLoS One 2018;13(2):e0191853.). In this study, we used RNA-seq to define up- and down regulated genes in both Ezh2 and G9a deficient (Math5Cre; Ezh2f/fG9af/+; dKO) retinal ganglion cells (RGC) to evaluate the hypothesis of Ezh2 and G9a interaction that has been discussed in other tissues but in the retina. ChIP-Seq was applied to evaluate H3K27me3 histone marks in retinal ganglion cells in wild type (WT), Ezh2 (Math5Cre; Ezh2-/-, sKO) and combined G9a-Ezh2 (Math5Cre; G9a+/-Ezh2-/-, dKO) knockout mutant mice at P1.

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