Journal Article

Defining the role of NG2-expressing cells in experimental models of multiple sclerosis. A biofunctional analysis of the neurovascular unit in wild type and NG2 null mice

Ferrara, Giovanni, Errede, Mariella, Annese, Tiziana, Morando, Sara, Longo, Giovanna, Trojano, Maria, Girolamo, Francesco, Alias, Carlotta, Uccelli, Antonio, Virgintino, Daniela, Kerlero de Rosbo, Nicole

2019

Overview

During experimental autoimmune encephalomyelitis (EAE), a model for multiple sclerosis associated with blood-brain barrier (BBB) disruption, oligodendrocyte precursor cells (OPCs) overexpress proteoglycan nerve/glial antigen 2 (NG2), proliferate, and make contacts with the microvessel wall. To explore whether OPCs may actually be recruited within the neurovascular unit (NVU), de facto intervening in its cellular and molecular composition, we quantified by immunoconfocal morphometry the presence of OPCs in contact with brain microvessels, during postnatal cerebral cortex vascularization at postnatal day 6, in wild-type (WT) and NG2 knock-out (NG2KO) mice, and in the cortex of adult naïve and EAE-affected WT and NG2KO mice. As observed in WT mice during postnatal development, a higher number of juxtavascular and perivascular OPCs was revealed in adult WT mice during EAE compared to adult naïve WT mice. In EAE-affected mice, OPCs were mostly associated with microvessels that showed altered claudin-5 and occludin tight junction (TJ) staining patterns and barrier leakage. In contrast, EAE-affected NG2KO mice, which did not show any significant increase in vessel-associated OPCs, seemed to retain better preserved TJs and BBB integrity. As expected, absence of NG2, in both OPCs and pericytes, led to a reduced content of vessel basal lamina molecules, laminin, collagen VI, and collagen IV. In addition, analysis of the major ligand/receptor systems known to promote OPC proliferation and migration indicated that vascular endothelial growth factor A (VEGF-A), platelet-derived growth factor-AA (PDGF-AA), and the transforming growth factor-β (TGF-β) were the molecules most likely involved in proliferation and recruitment of vascular OPCs during EAE. These results were confirmed by real time-PCR that showed Fgf2, Pdgfa and Tgfb expression on isolated cerebral cortex microvessels and by dual RNAscope-immunohistochemistry/in situ hybridization (IHC/ISH), which detected Vegfa and Vegfr2 transcripts on cerebral cortex sections. Overall, this study suggests that vascular OPCs, in virtue of their developmental arrangement and response to neuroinflammation and growth factors, could be integrated among the classical NVU cell components. Moreover, the synchronized activation of vascular OPCs and pericytes during both BBB development and dysfunction, points to NG2 as a key regulator of vascular interactions.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Angiogenesis / Animal models / Animals / Antigens / Antigens - biosynthesis / Antigens - genetics / Autoimmune diseases / Basal lamina / Biology and Life Sciences / Blood platelets / Blood-brain barrier / Blood-Brain Barrier - metabolism / Blood-Brain Barrier - pathology / Bone morphogenetic proteins / Brain / Brain research / Care and treatment / Cell adhesion & migration / Cell migration / Cell Movement - genetics / Cell proliferation / Cell Proliferation - genetics / Cerebral cortex / Cerebral Cortex - blood supply / Cerebral Cortex - metabolism / Cerebral Cortex - pathology / Chemical composition / Childrens health / Chondroitin sulfate / Claudin-5 - genetics / Claudin-5 - metabolism / Collagen / Collagen (type IV) / Disease Models, Animal / Disruption / Encephalomyelitis / Encephalomyelitis, Autoimmune, Experimental - genetics / Encephalomyelitis, Autoimmune, Experimental - metabolism / Encephalomyelitis, Autoimmune, Experimental - pathology / Endothelial growth factors / Endothelium / Experimental allergic encephalomyelitis / Fibroblast growth factor 2 / Fibroblast growth factors / Fibroblasts / Glial stem cells / Glycoproteins / Growth factors / Hybridization / Immunohistochemistry / Inflammation / Laminin / Medicine / Medicine and Health Sciences / Mice / Mice, Knockout / Microvessels - metabolism / Microvessels - pathology / Morphometry / Multiple sclerosis / Nervous system / Neurosciences / Oligodendroglia - metabolism / Oligodendroglia - pathology / Pericytes / Platelet-derived growth factor / Platelet-Derived Growth Factor - genetics / Platelet-Derived Growth Factor - metabolism / Proteoglycans / Proteoglycans - biosynthesis / Proteoglycans - genetics / Rehabilitation / Research and Analysis Methods / Risk factors / Stem Cells - metabolism / Stem Cells - pathology / Tight Junctions - genetics / Tight Junctions - metabolism / Tight Junctions - pathology / Transforming Growth Factor beta - genetics / Transforming Growth Factor beta - metabolism / Transforming growth factor-b / Transforming growth factors / Vascular endothelial growth factor / Vascular Endothelial Growth Factor A - genetics / Vascular Endothelial Growth Factor A - metabolism / Vascularization