Morphological and Functional Analysis of CNS-Associated Lymphatics

Methods Mol Biol. 2018:1846:141-151. doi: 10.1007/978-1-4939-8712-2_9.

Abstract

The study of meningeal lymphatic vessels of the central nervous system (CNS) has recently gathered momentum, with several papers dissecting their role in draining solutes from cerebrospinal fluid and brain (Louveau et al., Nature 523(7560):337-341, 2015; Antila et al., J Exp Med 214(12):3645-3667, 2017; Aspelund et al., J Exp Med 212(7):991-999, 2015). Methodological capabilities, however, have been limited to few laboratories due to difficulties reproducibly visualizing these rare cell subsets in the meninges. To explore meningeal lymphatics fundamental role during homeostasis and how they may contribute to human pathology, the field has begun to require purification and characterization of lymphatic endothelial cells. Here, modern cell biological techniques involving a combination of histological, flow-cytometric, and functional drainage assays are applied to brain and spinal cord meninges and detailed stepwise procedures used for successful in vivo and ex vivo characterization of meningeal lymphatic vessels.

Keywords: Brain meninges; CSF drainage; Neuroimmunology; Spinal cord meninges.

MeSH terms

  • Animals
  • Biomarkers
  • Central Nervous System / blood supply*
  • Central Nervous System / metabolism
  • Endothelial Cells / metabolism
  • Flow Cytometry
  • Immunohistochemistry
  • Lymph Nodes / blood supply
  • Lymph Nodes / metabolism
  • Lymphatic Vessels / anatomy & histology*
  • Lymphatic Vessels / metabolism*
  • Meninges / blood supply
  • Meninges / metabolism
  • Mice
  • Spinal Cord / blood supply
  • Spinal Cord / metabolism

Substances

  • Biomarkers