Tracking the evolution of CNS remyelinating lesion in mice with neutral red dye.
Animals
Astrocytes
/ drug effects
Cellular Microenvironment
/ drug effects
Central Nervous System
/ diagnostic imaging
Demyelinating Diseases
/ diagnostic imaging
Disease Models, Animal
Flow Cytometry
Humans
Lysophosphatidylcholines
/ toxicity
Mice
Microglia
/ drug effects
Multiple Sclerosis
/ diagnostic imaging
Myelin Sheath
/ drug effects
Nerve Regeneration
/ drug effects
Nervous System Diseases
/ diagnostic imaging
Neutral Red
/ pharmacology
Oligodendroglia
/ metabolism
Remyelination
/ drug effects
Spinal Cord Injuries
/ diagnostic imaging
animal models
demyelination
inflammation
remyelination
vital dye
Journal
Proceedings of the National Academy of Sciences of the United States of America
ISSN: 1091-6490
Titre abrégé: Proc Natl Acad Sci U S A
Pays: United States
ID NLM: 7505876
Informations de publication
Date de publication:
09 07 2019
09 07 2019
Historique:
pubmed:
27
6
2019
medline:
31
3
2020
entrez:
26
6
2019
Statut:
ppublish
Résumé
Animal models of central nervous system (CNS) demyelination, including toxin-induced focal demyelination and immune-mediated demyelination through experimental autoimmune encephalomyelitis (EAE), have provided valuable insights into the mechanisms of neuroinflammation and CNS remyelination. However, the ability to track changes in transcripts, proteins, and metabolites, as well as cellular populations during the evolution of a focal lesion, has remained challenging. Here, we developed a method to label CNS demyelinating lesions by the intraperitoneal injection of a vital dye, neutral red (NR), into mice before killing. We demonstrate that NR-labeled lesions can be easily identified on the intact spinal cord in both lysolecithin- and EAE-mediated demyelination models. Using fluorescence microscopy, we detected NR in activated macrophages/microglia and astrocytes, but not in oligodendrocytes present in lesions. Importantly, we successfully performed RT-qPCR, Western blot, flow cytometry, and mass spectrometry analysis of precisely dissected NR-labeled lesions at 5, 10, and 20 d postlesion (dpl) and found differential changes in transcripts, proteins, cell populations, and metabolites in lesions over the course of remyelination. Therefore, NR administration is a simple and powerful method to track and analyze the detailed molecular, cellular, and metabolic changes that occur within the lesion microenvironment over time following CNS injury. Furthermore, this method can be used to identify molecular and metabolic pathways that regulate neuroinflammation and remyelination and facilitate the development of therapies to promote repair in demyelinating disorders such as multiple sclerosis.
Identifiants
pubmed: 31235582
pii: 1819343116
doi: 10.1073/pnas.1819343116
pmc: PMC6628798
doi:
Substances chimiques
Lysophosphatidylcholines
0
Neutral Red
261QK3SSBH
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
14290-14299Subventions
Organisme : NINDS NIH HHS
ID : R01 NS107523
Pays : United States
Informations de copyright
Copyright © 2019 the Author(s). Published by PNAS.
Déclaration de conflit d'intérêts
The authors declare no conflict of interest.
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