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Lauren Laboissonniere, Jillian Goetz, Greg Schwartz, Jeffrey Trimarchi; Single cell transcriptomics of defined retinal ganglion cell type. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):2242.
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© ARVO (1962-2015); The Authors (2016-present)
Despite studies characterizing the distinct morphologies of retinal ganglion cell subtypes, the transcriptomic profiles of many of these cells have remained elusive. Specifically, there has not been a definitive study that links morphological and functional properties of ganglion cells to their transcriptome. Our project aims to uncover the genetic networks operating within subsets of retinal ganglion cells identified by different characteristics, including their physiology and morphology.
Individual retinal ganglion cells were isolated from mouse retinas based upon morphology, physiology or fluorescent reporter gene expression. After isolating the RNA and amplifying the cDNA for each individual cell, the resulting cDNA was either hybridized to microarrays or analyzed using RNA sequencing (SMART-seq).
We have analyzed individual ganglion cells through the use of both microarray and RNA sequencing techniques. Gene clusters have been identified that correlate with specific morphologies/electrophysiological properties of ganglion cells. Both methods also revealed transcriptomic variation between individual cells, which we have validated using a combination of in situ hybridizations and immunohistochemistry.
The analysis of single cells allows for in-depth studies regarding the genetic profile responsible for a particular cell’s function. This project isolated mature ganglion cells through the use of genetically driven fluorescent proteins and via electrophysiological properties. Our studies have yielded multiple genetic networks corresponding to cell function that we will utilize in the future to better understand the unique roles of these cells and how retinal ganglion cells are specified during development.
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