Abstract
To examine how astrocyte activation is regulated at different phases of relapsing–remitting EAE, we performed an immunofluorescent analysis of the spinal cord using the anti-glial fibrillary acidic protein (GFAP) monoclonal antibody GA-5. In keeping with previous studies, gray matter astrocytes showed strongly increased GFAP expression during the peak phase of disease (14 days post-immunization), which remained elevated during the remission phase (21–28 days post-immunization). In sharp contrast, during the peak phase of disease, the GA-5 signal in sub-meningeal white matter transiently disappeared in areas containing high levels of infiltrating leukocytes, but during the remission phase, the GFAP signal was fully restored. Parallel staining of the same sections with a polyclonal GFAP antibody confirmed elevated GFAP expression in the gray matter but no loss of signal in white matter. Interestingly, loss of GA-5 signal in sub-meningeal white matter was strongly associated with vascular disruption as defined by extravascular fibrinogen leak and by glio-vascular uncoupling, as defined by dissociation of AQP4-positive astrocyte endfeet and CD31-positive blood vessels. GA-5-negative areas were also associated with demyelination. These findings demonstrate a novel staining pattern of a GFAP antibody during EAE progression and suggest that the GFAP epitope recognized by the GA-5 monoclonal antibody transiently disappears as white matter astrocytes undergo remodeling during the peak phase of EAE. They also suggest that the GA-5 antibody provides a novel tool to identify astrocyte remodeling in other neurological conditions
Data Availability
The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.
References
Ballabh P, Braun A, Nedergaard M (2004) The blood-brain barrier: an overview. Structure, regulation and clinical implications. Neurobiology of Disease 16:1–13
Brosnan CF, Raine CS (2013) The astrocyte in multiple sclerosis revisited. Glia 61:453–465
Candelario-Jalil E, Yang Y, Rosenberg GA (2009) Diverse roles of matrixmetalloproteinsases and tissue inhibitors of metalloproteinases in neuroinflammation and cerebral ischemia. Neuroscience 158:983–994
Doshi A, Chataway J (2017) Multiple sclerosis, a treatable disease. Clin Med (Lond) 17:530–536
Eilam R, Segal M, Malach R, Sela M, Arnon R, Aharoni R (2018) Astrocyte disruption of neurovascular communication is linked to cortical damage in an animal model of multiple sclerosis. Glia 66:1098–1117
ffrench-Constant C (1994) Pathogenesis of multiple sclerosis. Lancet 343:271–275
Janzer RC, Raff MC (1987) Astrocytes induce blood-brain barrier properties in endothelial cells. Nature 325:253–257
Kothavale A, Di Gregorio D, Somera FP, Smith ME (1995) GFAP mRNA fluctuates in synchrony with chronic relapsing EAE symptoms in SJL/J mice. Glia 14:216–224
Lassmann H (1998) Multiple sclerosis pathology. In: Compston A (ed) McAlpine’s multiple sclerosis, 3rd edn. Churchill Livingstone, London, pp 323–358
McRae BL, Kennedy MK, Tan LJ, Dal Canto MC, Picha KS, Miller SD (1992) Induction of active and adoptive relapsing experimental autoimmune encephalomyelitis (EAE) using an encephalitogenic epitope of proteolipid protein. J Neuroimmunol 38:229–240
McRae BL, Vanderlugt CL, Dal Canto MC, Miller SD (1995) Functional evidence for epitope spreading in the relapsing pathology of experimental autoimmune encephalomyelitis. J Exp Med 182:75–85
Milner R, Campbell IL (2002) Developmental regulation of b1 integrins during angiogenesis in the central nervous system. Mol Cell Neurosci 20:616–626
Nedergaard M, Ransom B, Goldman SA (2003) New roles for astrocytes: redefining the functional architecture of the brain. Trends Neurosci 26(523–30):523–530
Papadopoulos MC, Verkman AS (2007) Aquaporin-4 and brain edema. Pediatr Nephrol 22:778–784
Prineas JW, Lee S (2019) Multiple sclerosis: destruction and regeneration of astrocytes in acute lesions. J Neuropath and Exp Neurol 78:140–156
Ransom B, Behar T, Nedergaard M (2003) New roles for astrocytes (stars at last). Trends Neurosci 26:520–522
Ridet J, Malhotra S, Privat A, Gage F (1997) Reactive astrocytes: cellular and molecular cues to biological function. Trends Neurosci 20:570–577
Rosenberg GA (2002) Matrix metalloproteinases in neuroinflammation. Glia 39:279–291
Shih AY, Fernandes HB, Choi F, Kozoriz MG, Liu Y, Cowan CM, Klegeris A (2006) Policing the police: astrocytes modulate microglial activation. J Neurosci 26:3887–3888
Smith ME, Somera FP, Eng LF (1983) Immunocytochemical staining for glial fibrillary acidic protein and the metabolism of cytoskeletal proteins in experimental allergic encephalomyelitis. Brain Res 264:241–253
Tari M, Glabinski AR, Tuohy VK, Stoler MH, Estes ML, Ransohoff RM (1996) In situ hybridization analysis of glial fibrillary acidic protein mRNA reveals evidence of biphasic astrocyte activation during acute experimental autoimmune encephalomyelitis. Am J Path 148:889–896
Tichauer J, Saud K, von Bernhardi R (2007) Modulation by astrocytes of microglial cell-mediated neuroinflammation: effect on the activation of microglial signaling pathways. NeuroImmunoModulation 14:168–174
Trivino A, Ramirez JM, Ramirez AI, Salazar JJ, Garcia-Sanchez J (1992) Retinal perivascular astrocglia: an immunoperoxidase study. Vision Res 32:1601–1607
Voskuhl RR, Peterson RS, Song B, Ao Y, Morales LBJ, Tiwari-Woodruff S, Sofroniew MV (2009) Reactive astrocytes form scar-like perivascular barriers to leukocytes during adaptive immune inflammation of the CNS. J Neurosci 29:11511–11522
Wang W, Schulze CJ, Suarez-Pinzon WL, Dyck JRB, Sawicki G, Schulz R (2002) Intracellular action of matrix metalloproteinase-2 accounts for myocardial ischemia and reperfusion injury. Circulation 106:1543–1549
Xie Y, Mustafa A, Yerzhan A, Merzhakupova D, Yerlan P, Orakov AN, Wang X, Huang Y, Miao L (2017) Nuclear matrix metalloproteinases: functions resemble the evolution from the intracellular to the extracellular compartment. Cell Death Discov 3:17036
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This work was supported by the NIH R56 Grant No. NS095753.
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SKH established the EAE studies, analyzed clinical EAE progression, performed the histological analysis and contributed to drafting the manuscript. RM conceived of the study, assisted in interpreting the findings and drafted the manuscript. Both authors read and approved the final manuscript.
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Halder, S.K., Milner, R. The GFAP Monoclonal Antibody GA-5 Identifies Astrocyte Remodeling and Glio-Vascular Uncoupling During the Evolution of EAE. Cell Mol Neurobiol 42, 1615–1622 (2022). https://doi.org/10.1007/s10571-021-01049-8
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DOI: https://doi.org/10.1007/s10571-021-01049-8