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Pericytes are enigmatic cells that contribute significantly to central nervous system (CNS) homeostasis, blood–brain barrier (BBB) regulation, angiogenesis, and maintenance of neurovascular integrity. These versatile cells are associated with blood vessels, embedded within the vascular basal lamina (VBL), and closest to endothelial cells (ECs). Their multifaceted roles in CNS physiology, development, and pathology have been implicated in various neurological disorders, such as stroke, Alzheimer’s disease, and multiple sclerosis. Immunohistochemistry (IHC) coupled with confocal microscopy imaging has emerged as a powerful tool for elucidating spatial distribution, molecular phenotype, and functional properties of CNS pericytes. Herein, we describe a detailed protocol using alternatively anti-NG2 or anti-PDGFR antibodies, to identify pericytes on high-resolution immunofluorescence, allowing us to study the three-dimensional network of anatomical interactions between these cells and the surrounding CNS parenchyma.

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Unveiling Central Nervous System Pericytes: Immunofluorescence Protocol and Confocal Microscopy

  • 实验方案

  • 发表日期：

    2025年

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  • Antonio d’Amati,
  • Domenico Ribatti,
  • Daniela Virgintino,
  • Mariella Errede

摘要

Pericytes are enigmatic cells that contribute significantly to central nervous system (CNS) homeostasis, blood–brain barrier (BBB) regulation, angiogenesis, and maintenance of neurovascular integrity. These versatile cells are associated with blood vessels, embedded within the vascular basal lamina (VBL), and closest to endothelial cells (ECs). Their multifaceted roles in CNS physiology, development, and pathology have been implicated in various neurological disorders, such as stroke, Alzheimer’s disease, and multiple sclerosis. Immunohistochemistry (IHC) coupled with confocal microscopy imaging has emerged as a powerful tool for elucidating spatial distribution, molecular phenotype, and functional properties of CNS pericytes. Herein, we describe a detailed protocol using alternatively anti-NG2 or anti-PDGFR antibodies, to identify pericytes on high-resolution immunofluorescence, allowing us to study the three-dimensional network of anatomical interactions between these cells and the surrounding CNS parenchyma.