Molecular profiling of resident and infiltrating mononuclear phagocytes during rapid adult retinal degeneration using single-cell RNA sequencing

Kaitryn E. Ronning, Sarah J. Karlen, Eric B. Miller, Marie E Burns

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Neuroinflammation commonly accompanies neurodegeneration, but the specific roles of resident and infiltrating immune cells during degeneration remains controversial. Much of the difficulty in assessing myeloid cell-specific functions during disease progression arises from the inability to clearly distinguish between activated microglia and bone marrow-derived monocytes and macrophages in various stages of differentiation and activation within the central nervous system. Using an inducible model of photoreceptor cell death, we investigated the prevalence of infiltrating monocytes and macrophage subpopulations after the initiation of degeneration in the mouse retina. In vivo retinal imaging revealed infiltration of CCR2 + leukocytes across retinal vessels and into the parenchyma within 48 hours of photoreceptor degeneration. Immunohistochemistry and flow cytometry confirmed and characterized these leukocytes as CD11b + CD45 + cells. Single-cell mRNA sequencing of the entire CD11b + CD45 + population revealed the presence of resting microglia, activated microglia, monocytes, and macrophages as well as 12 distinct subpopulations within these four major cell classes. Our results demonstrate a previously immeasurable degree of molecular heterogeneity in the innate immune response to cell-autonomous degeneration within the central nervous system and highlight the necessity of unbiased high-throughput and high-dimensional molecular techniques like scRNAseq to understand the complex and changing landscape of immune responders during disease progression.

Original languageEnglish (US)
Article number4858
JournalScientific Reports
Volume9
Issue number1
DOIs
StatePublished - Dec 1 2019

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RNA Sequence Analysis
Retinal Degeneration
Phagocytes
Microglia
Macrophages
Disease Progression
Monocytes
Leukocytes
Central Nervous System
Retinal Vessels
Photoreceptor Cells
Myeloid Cells
Innate Immunity
Retina
Flow Cytometry
Cell Death
Immunohistochemistry
Messenger RNA
Population

ASJC Scopus subject areas

  • General

Cite this

Molecular profiling of resident and infiltrating mononuclear phagocytes during rapid adult retinal degeneration using single-cell RNA sequencing. / Ronning, Kaitryn E.; Karlen, Sarah J.; Miller, Eric B.; Burns, Marie E.

In: Scientific Reports, Vol. 9, No. 1, 4858, 01.12.2019.

Research output: Contribution to journalArticle

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