Methods Mol Biol. 2023;2636:19-41. doi: 10.1007/978-1-0716-3012-9_2.
We describe a computational workflow to analyze single-cell RNA-sequencing (scRNA-seq) profiles of axotomized retinal ganglion cells (RGCs) in mice. Our goal is to identify differences in the dynamics of survival among 46 molecularly defined RGC types together with molecular signatures that correlate with these differences. The data consists of scRNA-seq profiles of RGCs collected at six time points following optic nerve crush (ONC) (see companion chapter by Jacobi and Tran). We use a supervised classification-based approach to map injured RGCs to type identities and quantify type-specific differences in survival at 2 weeks post crush. As injury-related changes in gene expression confound the inference of type identity in surviving cells, the approach deconvolves type-specific gene signatures from injury responses by using an iterative strategy that leverages measurements along the time course. We use these classifications to compare expression differences between resilient and susceptible subpopulations, identifying potential mediators of resilience. The conceptual framework underlying the method is sufficiently general for analysis of selective vulnerability in other neuronal systems.
PMID:36881293 | DOI:10.1007/978-1-0716-3012-9_2