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Genetic signatures of retinal ganglion cells revealed through single cell profiling

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Mus musculus

PROVIDER: GSE115380 | GEO | 2019/11/01

REPOSITORIES: GEO

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Molecular signatures of retinal ganglion cells revealed through single cell profiling.

Project description:Retinal ganglion cells can be classified into more than 40 distinct subtypes, whether by functional classification or transcriptomics. The examination of these subtypes in relation to their physiology, projection patterns, and circuitry would be greatly facilitated through the identification of specific molecular identifiers for the generation of transgenic mice. Advances in single cell transcriptomic profiling have enabled the identification of molecular signatures for cellular subtypes that are only rarely found. Therefore, we used single cell profiling combined with hierarchical clustering and correlate analyses to identify genes expressed in distinct populations of Parvalbumin-expressing cells and functionally classified RGCs. RGCs were manually isolated based either upon fluorescence or physiological distinction through cell-attached recordings. Microarray hybridization and RNA-Sequencing were employed for the characterization of transcriptomes and in situ hybridization was utilized to further characterize gene candidate expression. Gene candidates were identified based upon cluster correlation, as well as expression specificity within physiologically distinct classes of RGCs. Further, we identified Prph, Ctxn3, and Prkcq as potential candidates for ipRGC classification in the murine retina. The use of these genes, or one of the other newly identified subset markers, for the generation of a transgenic mouse would enable future studies of RGC-subtype specific function, wiring, and projection.

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Genetic signatures of retinal ganglion cells revealed through single cell profiling [dissociated RGC]

Project description:This study aimed to characterize the transcriptomes of adult retinal ganglion cells (RGCs). These cells were isolated through their expression of a TdTomato molecule that would only turn on when Cre was present. Cre was controlled by the Parvalbumin promoter.

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Genetic signatures of retinal ganglion cells revealed through single cell profiling [non-dissociated RGC]

Project description:This study aimed to characterize the transcriptomes of adult retinal ganglion cells (RGCs). The physiological properties of these cells were characterized first in non-dissociated retinal tissue. With that information recorded, the cells were isolated for transcriptome profiling.

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Genetic signatures of retinal ganglion cells revealed through single cell profiling

Project description:Genetic signatures of retinal ganglion cells revealed through single cell profiling

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Genetic signatures of retinal ganglion cells revealed through single cell profiling [dissociated RGC]

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Genetic signatures of retinal ganglion cells revealed through single cell profiling [non-dissociated RGC]

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Disease gene candidates revealed by expression profiling of retinal ganglion cell development.

Project description:To what extent do postmitotic neurons regulate gene expression during development or after injury? We took advantage of our ability to highly purify retinal ganglion cells (RGCs) to profile their pattern of gene expression at 13 ages from embryonic day 17 through postnatal day 21. We found that a large proportion of RGC genes are regulated dramatically throughout their postmitotic development, although the genes regulated through development in vivo generally are not regulated similarly by RGCs allowed to age in vitro. Interestingly, we found that genes regulated by developing RGCs are not generally correlated with genes regulated in RGCs stimulated to regenerate their axons. We unexpectedly found three genes associated with glaucoma, optineurin, cochlin, and CYP1B1 (cytochrome P450, family 1, subfamily B, polypeptide 1), previously thought to be primarily expressed in the trabecular meshwork, which are highly expressed by RGCs and regulated through their development. We also identified several other RGC genes that are encoded by loci linked to glaucoma. The expression of glaucoma-linked genes by RGCs suggests that, at least in some cases, RGCs may be directly involved in glaucoma pathogenesis rather than indirectly involved in response to increased intraocular pressure. Consistent with this hypothesis, we found that CYP1B1 overexpression potentiates RGC survival.

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Single cell transcriptome profiling of retinal ganglion cells identifies cellular subtypes.

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