Faculty Collaboration Database

LCM-Seq for Retinal Cell Layer-Specific Responses During Optic Nerve Regeneration. Methods Mol Biol 2023;2636:311-321

Date

03/08/2023

Pubmed ID

36881308

DOI

10.1007/978-1-0716-3012-9_17

Scopus ID

2-s2.0-85149523514 (requires institutional sign-in at Scopus site)

Abstract

LCM-seq is a powerful tool for gene expression analysis from individual or groups of cells that can be spatially isolated. Within the visual system, retinal ganglion cells (RGCs), the cells that connect the eye to the brain through the optic nerve, reside in the retinal ganglion cell layer of the retina. This well-defined location provides a unique opportunity to harvest RNA by laser capture microdissection (LCM) from a highly enriched cell population. Using this method, it is possible to explore transcriptome-wide changes in gene expression following optic nerve injury. In the zebrafish model, this method can be used to identify molecular events driving successful optic nerve regeneration in contrast to mammals that fail to regenerate axons in the central nervous system. Here we provide a method for LCM from the different retinal layers of zebrafish following optic nerve injury and during the process of optic nerve regeneration. Purified RNA from this protocol is sufficient for RNA-seq or other downstream analysis.

Author List

Speer W, Veldman MB

Author

Matthew B. Veldman PhD Assistant Professor in the Cell Biology, Neurobiology and Anatomy department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Animals
Laser Capture Microdissection
Mammals
Nerve Regeneration
Optic Nerve Injuries
RNA
Retinal Ganglion Cells
Zebrafish