The mouse cortical meninges are the site of immune responses to many different pathogens, and are accessible to intravital imaging

Coles, J. A., Stewart-Hutchinson, P. J., Myburgh, E. and Brewer, J. M. (2017) The mouse cortical meninges are the site of immune responses to many different pathogens, and are accessible to intravital imaging. Methods, 127, pp. 53-61. (doi: 10.1016/j.ymeth.2017.03.020) (PMID:28351758) (PMCID:PMC5595162)

[img]
Preview
Text
138992.pdf - Published Version
Available under License Creative Commons Attribution.

2MB

Abstract

A wide range of viral and microbial infections are known to cause meningitis, and there is evidence that the meninges are the gateway to pathogenic invasion of the brain parenchyma. Hence observation of these regions has wide application to understanding host-pathogen interactions. Interactions between pathogens and cells of the immune response can be modified by changes in their environment, such as suppression of the flow of blood and lymph, and, particularly in the case of the meninges, with their unsupported membranes, invasive dissection can alter the tissue architecture. For these reasons, intravital imaging through the unperforated skull is the method of choice. We give a protocol for a simple method of two-photon microscopy through the thinned cortical skull of the anesthetized mouse to enable real-time imaging with sub-micron resolution through the meninges and into the superficial brain parenchyma. In reporter mice in which selected cell types express fluorescent proteins, imaging after infection with fluorescent pathogens (lymphocytic choriomeningitis virus, Trypanosoma brucei or Plasmodium berghei) has shown strong recruitment to the cortical meninges of immune cells, including neutrophils, T cells, and putative dendritic cells and macrophages. Without special labeling, the boundaries between the dura mater, the leptomeninx, and the parenchyma are not directly visualized in intravital two-photon microscopy, but other landmarks and characteristics, which we illustrate, allow the researcher to identify the compartment being imaged. While most infectious meningitides are localized mainly in the dura mater, others involve recruitment of immune cells to the leptomeninx.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Brewer, Professor James and Coles, Dr Jonathan and Myburgh, Dr Elmarie
Authors: Coles, J. A., Stewart-Hutchinson, P. J., Myburgh, E., and Brewer, J. M.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Methods
Publisher:Elsevier
ISSN:1046-2023
ISSN (Online):1095-9130
Published Online:27 March 2017
Copyright Holders:Copyright © 2017 The Authors
First Published:First published in Methods 127:53-61
Publisher Policy:Reproduced under a Creative Commons License

University Staff: Request a correction | Enlighten Editors: Update this record

Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
522031In vivo imaging models of African trypanosomiasis to support drug discovery programmes.Michael BarrettGates Foundation (GATES)OPPGH5337III - PARASITOLOGY
371796The Wellcome Centre for Molecular Parasitology ( Core Support )Andrew WatersWellcome Trust (WELLCOME)085349/Z/08/ZIII - PARASITOLOGY