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MicroRNA expression profile in mouse cone photoreceptors at P60 [miRNA-seq]

ABSTRACT: The outer segments of cones serve as light detectors for daylight color vision, and their dysfunction leads to human blindness conditions. We show that the cone-specific disruption of DiGeorge Syndrome Critical Region Gene 8 (DGCR8) in adult mice led to the loss of miRNAs and the loss of outer segments, resulting in photoreceptors with significantly reduced light responses. Using next-generation sequencing of RNA from isolated wild type P60 cones, we determine the most highly expressed miRNAs as candidates for controlling outer segment maintenance. The expression pattern of miRNAs was highly uneven, with a single miRNA, miR-182, representing 64% of all miRNA expressed in cones. Re-expression of miR-182 and miR-183 (third most abundant miRNA) prevented outer segment loss. These miRNAs were also necessary and sufficient for the formation of inner segments, connecting cilia and short outer segments, as well as light responses in stem-cell-derived retinal cultures. Our results show that miR-182- and miR-183-regulated pathways are necessary for cone outer segment maintenance in vivo and functional outer segment formation in vitro. microRNA profile in cone photoreceptors from P60 D4-cre/Ai9 tdTomato mice representing wild type control.

ORGANISM(S): Mus musculus

SUBMITTER: Tim Roloff

PROVIDER: E-GEOD-58500 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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MicroRNA expression profile in mouse cone photoreceptors at P60 [miRNA-seq]

Project description:The outer segments of cones serve as light detectors for daylight color vision, and their dysfunction leads to human blindness conditions. We show that the cone-specific disruption of DiGeorge Syndrome Critical Region Gene 8 (DGCR8) in adult mice led to the loss of miRNAs and the loss of outer segments, resulting in photoreceptors with significantly reduced light responses. Using next-generation sequencing of RNA from isolated wild type P60 cones, we determine the most highly expressed miRNAs as candidates for controlling outer segment maintenance. The expression pattern of miRNAs was highly uneven, with a single miRNA, miR-182, representing 64% of all miRNA expressed in cones. Re-expression of miR-182 and miR-183 (third most abundant miRNA) prevented outer segment loss. These miRNAs were also necessary and sufficient for the formation of inner segments, connecting cilia and short outer segments, as well as light responses in stem-cell-derived retinal cultures. Our results show that miR-182- and miR-183-regulated pathways are necessary for cone outer segment maintenance in vivo and functional outer segment formation in vitro.

2014-06-20 | GSE58500 | GEO

Expression data of wild type and C-DGCR8 KO cones at different timepoints [RNA-seq]

Project description:We have analyzed gene expression in cone photoreceptors isolated from wild type and C-DGCR8 (DiGeorge Syndrome Critical Region Gene 8) KO mice at five different time points to get a mechanistic inside into the altered molecular pathways after microRNAs depletion. Cones were isolated by FACS from wild type and C-DGCR8 KO mice expressing Cre recombinase postnatally specifically in cones (D4-Cre) and bearing conditional null Dgcr8 allele. Experiments were performed in duplicates from different time points (P30, P40, P50, P60, and P90). Total RNA was extracted and next-generation RNA sequencing was performed.

2014-06-20 | E-GEOD-58498 | biostudies-arrayexpress

Expression data of wild type and C-DGCR8 KO cones at different timepoints [array]

2014-06-20 | E-GEOD-58493 | biostudies-arrayexpress

Rab28 is localised to photoreceptor outer segments, regulates outer segment shedding but is not linked to retinal degeneration in zebrafish

Project description:The photoreceptor outer segment is the canonical example of a modified and highly specialised cilium, with an expanded membrane surface area in the form of discs or lamellae for efficient light detection. Many ciliary proteins are essential for normal photoreceptor function and cilium dysfunction often results in retinal degeneration leading to impaired vision. Herein, we investigate the function and localisation of the ciliary G-protein RAB28 in zebrafish cone photoreceptors. CRISPR-Cas9 generated rab28 mutant zebrafish display a reduction in shed outer segment material in the RPE at 1 month post fertilisation (mpf), but otherwise normal retinal structure and visual function up to 12 mpf. Cone photoreceptorspecific transgenic reporter lines show Rab28 localises almost exclusively to outer segments, independently of nucleotide binding. Co-immunoprecipitation analysis demonstrates tagged Rab28 interacts with components of the phototransduction cascade, including opsins, Phosphodiesterase 6C and Guanylate Cyclase 2D. Our data shed light on RAB28 function in cones and provide a model for RAB28-associated cone-rod dystrophy

2020-04-09 | PXD017523 | Pride

MiRNAs 182 and 183 are necessary to maintain adult cone photoreceptor outer segments and visual function

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

2014-06-20 | E-GEOD-58501 | biostudies-arrayexpress

Proteomic analysis of growth cones from WT and HD mice

Project description:Although Huntington’s disease (HD) is a late onset neurological condition, studies suggest a developmental contribution to its adult manifestations. Imaging studies in presymptomatic HD gene carriers revealed early alterations in white matter tract with a marked defect in the corpus callosum. This tract is mainly formed by axons projecting from layer II/III neurons. HD leads to microtubule bundling defects in the growth cone, the cellular compartment that drives axonal growth. We performed a mass spectrometry-based proteomic analysis of growth cones from WT (HdhQ7/Q7) and HD (HdhQ7/Q111) mice to identify proteins involved in HD-induced phenotypes on axonal growth and on the growth cone microtubule cytoskeleton.

2021-12-08 | PXD023885 | Pride

Reactivating the phototransduction cascade with a mutation agnostic gene therapy preserves vision in Rod-Cone Dystrophies

Project description:Rod-cone dystrophy (RCD), or retinitis pigmentosa, involves genetic conditions where rod degeneration precedes cone degeneration, resulting in tunnel vision and eventual blindness. Previous studies attempted to restore cone activity and prolong vision by expressing microbial chloride pumps in degenerating cone photoreceptors. However, microbial opsins require high expression and intense light, limiting their effectiveness. In this work, we explored the phototransduction cascade in degenerating cones in two RCD mouse models. We observed that opsin and arrestin expression persists in the soma during outer segment degeneration. This led us to hypothesize that reactivating cones based on cone opsin signaling may be possible by expressing a target channel activated by G proteins recruited by cone opsin. Through adeno-associated viral (AAV) mediated expression of the G protein-coupled inwardly rectifying K (GIRK) channel, we achieved improvements in visual function in two RCD mouse models with mutations in different genes. Importantly, we validated the rationale of GIRK-mediated gene therapy in humans by confirming cone opsin and cone arrestin expression in cones of late-stage RCD patients. Our proposed approach involves GIRK channel expression in cones as a novel method to maintain high acuity, sensitivity, and color vision in RCD, independent of the underlying mutation.

2025-01-13 | GSE286529 | GEO

Single-cell RNASeq analysis of human retinal organoids generated from hiPSC derived of a healthy donor patient

Project description:Results: Our histologic studies indicated that human retinal organoids (HROs) at day 200 of differentiation in this system are postmitotic and thus completed retinogenesis. Further, HRO contain all major retinal cell types in a laminated structure. Notably, HROs are cone photoreceptor-rich, show a 1:1:1 ratio of Müller glia, rod and cone photoreceptor. Immunostaining and ultrastructural studies showed that photoreceptors neurons mature, including photoreceptor inner and nascent outer segment formation. Our transcriptome analysis at the single cell level supports these findings. Further, comparison with published datasets (Voigt et al. 2019 [PMID: 31075224]) indicate that the genotype of cone photoreceptors in this HRO system correlates more strongly with the foveal cones of the human primary retina than peripheral cones. Müller glia show a trend towards human fovea whereas rod photoreceptors were found to be nearly similar. Conclusions: Single cell transcriptome analysis of HROs support and extend our findings at the histological level, indicating that HROs are cone photoreceptor-rich, and provide some characteristics of the human macula.

2022-09-01 | GSE174215 | GEO

miR-182 regulates Slit2-mediated axon guidance by modulating the local translation of a specific mRNA

Project description:During brain wiring, mRNAs are trafficked into axons and growth cones where they are differentially translated in response to extrinsic signals. Differential control of local protein synthesis mediates neuronal compartment-specific behaviors that aid axon guidance. Yet little is understood about how specific mRNAs are selected for translation. Here we have investigated the local role of microRNAs (miRNAs) in mRNA-specific translation during axon pathfinding of Xenopus laevis retinal ganglion cell (RGC) axons. Profiling experiments revealed a rich repertoire of axonal miRNAs in developing RGC axons and identified miR-182 as one of the most abundant. Loss of miR182 impairs Slit2-induced growth cone repulsion and causes RGC axon targeting defects in vivo. To aid miRNA target prediction, we also profiled mRNA expression in RGC axons. Our results show that miR-182 targets cofilin1 mRNA in RGC growth cones and modulates its local translation in response to Slit2.

2017-01-23 | GSE86883 | GEO

A transcriptomic resource for Douglas-fir seed development and analysis of transcription during late megagametophyte development

Project description:We developed a transcriptome resource for Douglas-fir covering key developmental stages of megagametophytes over time: prefertilization, fertilization, embryogenesis, and early, unfertilized abortion. Extracted RNA was sequenced using large-scale sequencing and reads were assembled to generate a de novo reference transcriptome of 105,505 predicted high-confidence transcripts. Expression levels were estimated based on alignment of the original reads to the reference. 200â400 megagametophytes were dissected and pooled per sample on four dates from either pollinated or unpollinated cones: June 10, June 22, June 30, and July 6 2011. These dates coincided with key events in seed development: corrosion cavity formation, fertilization, embryogenesis, or the early stages of abortion in the unpollinated treatment. Sporophytic tissue (i.e. cone bracts and cone scales) were added for comparison. PolyA RNA was used for Illumina sequencing.

2016-06-16 | E-GEOD-83425 | biostudies-arrayexpress

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