Abstract
Microglia represent resident immune cells of the central nervous system (CNS), which have been shown to be involved in the pathophysiology of practically every neuropathology. As microglia were described to participate in the formation of the astroglial glia limitans around CNS vessels, they are part of the neurovascular unit (NVU). Since the NVU is a highly specialized structure, being functionally and morphologically adapted to differing demands in the arterial, capillary, and venous segments, the present study was aimed to systematically investigate the microglial contribution to the glia limitans along the vascular tree. Thereby, the microglial participation in the glia limitans was demonstrated for arteries, capillaries, and veins by immunoelectron microscopy in wild-type mice. Furthermore, analysis by confocal laser scanning microscopy revealed the highest density of microglial endfeet contacting the glial basement membrane around capillaries, with significantly lower densities around arteries and veins. Importantly, this pattern appeared to be unaltered in the setting of experimental autoimmune encephalomyelitis (EAE) in CX3CR1CreERT2:R26-Tomato reporter mice, although perivascular infiltrates of blood-borne leukocytes predominantly occur at the level of post-capillary venules. However, EAE animals exhibited significantly increased contact sizes of individual microglial endfeet around arteries and veins. Noteworthy, under EAE conditions, the upregulation of MHC-II was not limited to microglia of the glia limitans of veins showing infiltrates of leukocytes, but also appeared at the capillary level. As a microglial contribution to the glia limitans was also observed in human brain tissue, these findings may help characterizing microglial alterations within the NVU in various neuropathologies.
Similar content being viewed by others
References
Agrawal S, Anderson P, Durbeej M, van Rooijen N, Ivars F, Opdenakker G, Sorokin LM (2006) Dystroglycan is selectively cleaved at the parenchymal basement membrane at sites of leukocyte extravasation in experimental autoimmune encephalomyelitis. J Exp Med 203:1007–1019. https://doi.org/10.1084/jem.20051342
Ajami B, Bennett JL, Krieger C, Tetzlaff W, Rossi FMV (2007) Local self-renewal can sustain CNS microglia maintenance and function throughout adult life. Nat Neurosci 10:1538–1543. https://doi.org/10.1038/nn2014
Alliot F, Godin I, Pessac B (1999) Microglia derive from progenitors, originating from the yolk sac, and which proliferate in the brain. Dev Brain Res 117:145–152. https://doi.org/10.1016/S0165-3806(99)00113-3
Askew K, Li K, Olmos-Alonso A, Garcia-Moreno F, Liang Y, Richardson P, Tipton T, Chapman MA, Riecken K, Beccari S, Sierra A, Molnár Z, Cragg MS, Garaschuk O, Perry VH, Gomez-Nicola D (2017) Coupled proliferation and apoptosis maintain the rapid turnover of microglia in the adult brain. Cell Rep 18:391–405. https://doi.org/10.1016/j.celrep.2016.12.041
Barkauskas DS, Evans TA, Myers J, Petrosiute A, Silver J, Huang AY (2013) Extravascular CX3CR1 + cells extend intravascular dendritic processes into intact central nervous system vessel lumen. Microsc Microanal 19:778–790. https://doi.org/10.1017/S1431927613000482
Bechmann I, Kwidzinski E, Kovac AD, Simbürger E, Horvath T, Gimsa U, Dirnagl U, Priller J, Nitsch R (2001) Turnover of rat brain perivascular cells. Exp Neurol 168:242–249. https://doi.org/10.1006/exnr.2000.7618
Bechmann I, Goldmann J, Kovac AD, Kwidzinski E, Simbürger E, Naftolin F, Dirnagl U, Nitsch R, Priller J (2005) Circulating monocytic cells infiltrate layers of anterograde axonal degeneration where they transform into microglia. FASEB J 19:647–649. https://doi.org/10.1096/fj.04-2599fje
Bechmann I, Galea I, Perry VH (2007) What is the blood–brain barrier (not)? Trends Immunol 28:5–11. https://doi.org/10.1016/j.it.2006.11.007
Bennett ML, Bennett FC, Liddelow SA, Ajami B, Zamanian JL, Fernhoff NB, Mulinyawe SB, Bohlen CJ, Adil A, Tucker A, Weissman IL, Chang EF, Li G, Grant GA, Hayden Gephart MG, Barres BA (2016) New tools for studying microglia in the mouse and human CNS. Proc Natl Acad Sci USA 113:E1738–E1746. https://doi.org/10.1073/pnas.1525528113
Colonna M, Butovsky O (2017) Microglia function in the central nervous system during health and neurodegeneration. Annu Rev Immunol 35:441–468. https://doi.org/10.1146/annurev-immunol-051116-052358
Dyrna F, Hanske S, Krueger M, Bechmann I (2013) The blood-brain barrier. J Neuroimmune Pharmacol 8:763–773. https://doi.org/10.1007/s11481-013-9473-5
Ebner F, Brandt C, Thiele P, Richter D, Schliesser U, Siffrin V, Schueler J, Stubbe T, Ellinghaus A, Meisel C, Sawitzki B, Nitsch R (2013) Microglial activation milieu controls regulatory T cell responses. J Immunol 191:5594–5602. https://doi.org/10.4049/jimmunol.1203331
Edvinsson L, Högestätt ED, Uddman R, Auer LM (1983) Cerebral veins: fluorescence histochemistry, electron microscopy, and in vitro reactivity. J Cereb Blood Flow Metab 3(2):226–230
Engelhardt B, Carare RO, Bechmann I, Flugel A, Laman JD, Weller RO (2016) Vascular, glial, and lymphatic immune gateways of the central nervous system. Acta Neuropathol 132:317–338. https://doi.org/10.1007/s00401-016-1606-5
Geissmann F, Gordon S, Hume DA, Mowat AM, Randolph GJ (2010) Unravelling mononuclear phagocyte heterogeneity. Nat Rev Immunol 10:453–460. https://doi.org/10.1038/nri2784
Gertig U, Hanisch U-K (2014) Microglial diversity by responses and responders. Front Cell Neurosci 8:101. https://doi.org/10.3389/fncel.2014.00101
Ginhoux F, Greter M, Leboeuf M, Nandi S, See P, Gokhan S, Mehler MF, Conway SJ, Ng LG, Stanley ER, Samokhvalov IM, Merad M (2010) Fate mapping analysis reveals that adult microglia derive from primitive macrophages. Science 330:841–845. https://doi.org/10.1126/science.1194637
Goldmann T, Wieghofer P, Müller PF, Wolf Y, Varol D, Yona S, Brendecke SM, Kierdorf K, Staszewski O, Datta M, Luedde T, Heikenwalder M, Jung S, Prinz M (2013) A new type of microglia gene targeting shows TAK1 to be pivotal in CNS autoimmune inflammation. Nat Neurosci 16:1618–1626. https://doi.org/10.1038/nn.3531
Goldmann T, Wieghofer P, Jordao MJC, Prutek F, Hagemeyer N, Frenzel K, Amann L, Staszewski O, Kierdorf K, Krueger M, Locatelli G, Hochgerner H, Zeiser R, Epelman S, Geissmann F, Priller J, Rossi FMV, Bechmann I, Kerschensteiner M, Linnarsson S, Jung S, Prinz M (2016) Origin, fate and dynamics of macrophages at central nervous system interfaces. Nat Immunol 17:797–805. https://doi.org/10.1038/ni.3423
Hannocks M-J, Zhang X, Gerwien H, Chashchina A, Burmeister M, Korpos E, Song J, Sorokin L (2017) The gelatinases, MMP-2 and MMP-9, as fine tuners of neuroinflammatory processes. Matrix Biol. https://doi.org/10.1016/j.matbio.2017.11.007
Hanske S, Dyrna F, Bechmann I, Krueger M (2016) Different segments of the cerebral vasculature reveal specific endothelial specifications, while tight junction proteins appear equally distributed. Brain Struct Funct. https://doi.org/10.1007/s00429-016-1267-0
Hawkes CA, Härtig W, Kacza J, Schliebs R, Weller RO, Nicoll JA, Carare RO (2011) Perivascular drainage of solutes is impaired in the ageing mouse brain and in the presence of cerebral amyloid angiopathy. Acta Neuropathol 121:431–443. https://doi.org/10.1007/s00401-011-0801-7
Hefendehl JK, Neher JJ, Sühs RB, Kohsaka S, Skodras A, Jucker M (2014) Homeostatic and injury-induced microglia behavior in the aging brain. Aging Cell 13:60–69. https://doi.org/10.1111/acel.12149
Hell SW, Wichmann J (1994) Breaking the diffraction resolution limit by stimulated emission: Stimulated-emission-depletion fluorescence microscopy. Opt Lett 19:780–782
Immig K, Gericke M, Menzel F, Merz F, Krueger M, Schiefenhovel F, Losche A, Jager K, Hanisch U-K, Biber K, Bechmann I (2015) CD11c-positive cells from brain, spleen, lung, and liver exhibit site-specific immune phenotypes and plastically adapt to new environments. Glia 63:611–625. https://doi.org/10.1002/glia.22771
Jolivel V, Bicker F, Binamé F, Ploen R, Keller S, Gollan R, Jurek B, Birkenstock J, Poisa-Beiro L, Bruttger J, Opitz V, Thal SC, Waisman A, Bäuerle T, Schäfer MK, Zipp F, Schmidt MHH (2015) Perivascular microglia promote blood vessel disintegration in the ischemic penumbra. Acta Neuropathol 129:279–295. https://doi.org/10.1007/s00401-014-1372-1
Jung S, Aliberti J, Graemmel P, Sunshine MJ, Kreutzberg GW, Sher A, Littman DR (2000) Analysis of fractalkine receptor CX3CR1 function by targeted deletion and green fluorescent protein reporter gene insertion. Mol Cell Biol 20:4106–4114. https://doi.org/10.1128/MCB.20.11.4106-4114.2000
Kierdorf K, Prinz M (2017) Microglia in steady state. J Clin Invest 127:3201–3209. https://doi.org/10.1172/JCI90602
Kierdorf K, Erny D, Goldmann T, Sander V, Schulz C, Perdiguero EG, Wieghofer P, Heinrich A, Riemke P, Hölscher C, Müller DN, Luckow B, Brocker T, Debowski K, Fritz G, Opdenakker G, Diefenbach A, Biber K, Heikenwalder M, Geissmann F, Rosenbauer F, Prinz M (2013) Microglia emerge from erythromyeloid precursors via Pu.1- and Irf8-dependent pathways. Nat Neurosci 16:273–280. https://doi.org/10.1038/nn.3318
Krueger M, Härtig W, Frydrychowicz C, Mueller WC, Reichenbach A, Bechmann I, Michalski D (2017) Stroke-induced blood-brain barrier breakdown along the vascular tree - No preferential affection of arteries in different animal models and in humans. J Cereb Blood Flow Metab 37:2539–2554. https://doi.org/10.1177/0271678X16670922
Lassmann H, Bradl M (2017) Multiple sclerosis: Experimental models and reality. Acta Neuropathol 133:223–244. https://doi.org/10.1007/s00401-016-1631-4
Lassmann H, Zimprich F, Vass K, Hickey WF (1991) Microglial cells are a component of the perivascular glia limitans. J Neurosci Res 28:236–243. https://doi.org/10.1002/jnr.490280211
Lawson LJ, Perry VH, Dri P, Gordon S (1990) Heterogeneity in the distribution and morphology of microglia in the normal adult mouse brain. Neuroscience 39:151–170. https://doi.org/10.1016/0306-4522(90)90229-W
Liebner S, Dijkhuizen RM, Reiss Y, Plate KH, Agalliu D, Constantin G (2018) Functional morphology of the blood-brain barrier in health and disease. Acta Neuropathol 135:311–336. https://doi.org/10.1007/s00401-018-1815-1
Mildner A, Schmidt H, Nitsche M, Merkler D, Hanisch U-K, Mack M, Heikenwalder M, Bruck W, Priller J, Prinz M (2007) Microglia in the adult brain arise from Ly-6ChiCCR2 + monocytes only under defined host conditions. Nat Neurosci 10:1544–1553. https://doi.org/10.1038/nn2015
Mildner A, Huang H, Radke J, Stenzel W, Priller J (2017) P2Y12 receptor is expressed on human microglia under physiological conditions throughout development and is sensitive to neuroinflammatory diseases. Glia 65:375–387. https://doi.org/10.1002/glia.23097
Mittelbronn M, Dietz K, Schluesener HJ, Meyermann R (2001) Local distribution of microglia in the normal adult human central nervous system differs by up to one order of magnitude. Acta Neuropathol 101:249–255
O’Loughlin E, Madore C, Lassmann H, Butovsky O (2018) Microglial phenotypes and functions in multiple sclerosis. Cold Spring Harb Perspect Med 8. https://doi.org/10.1101/cshperspect.a028993
Poliani PL, Wang Y, Fontana E, Robinette ML, Yamanishi Y, Gilfillan S, Colonna M (2015) TREM2 sustains microglial expansion during aging and response to demyelination. J Clin Invest 125:2161–2170. https://doi.org/10.1172/JCI77983
Prodinger C, Bunse J, Kruger M, Schiefenhovel F, Brandt C, Laman JD, Greter M, Immig K, Heppner F, Becher B, Bechmann I (2011) CD11c-expressing cells reside in the juxtavascular parenchyma and extend processes into the glia limitans of the mouse nervous system. Acta Neuropathol 121:445–458. https://doi.org/10.1007/s00401-010-0774-y
Rodrigues MCO, Hernandez-Ontiveros DG, Louis MK, Willing AE, Borlongan CV, Sanberg PR, Voltarelli JC, Garbuzova-Davis S (2012) Neurovascular aspects of amyotrophic lateral sclerosis. Int Rev Neurobiol 102:91–106. https://doi.org/10.1016/B978-0-12-386986-9.00004-1
Schetters STT, Gomez-Nicola D, Garcia-Vallejo JJ, van Kooyk Y (2017) Neuroinflammation: microglia and T Cells get ready to tango. Front Immunol 8:1905. https://doi.org/10.3389/fimmu.2017.01905
Sierra A, Gottfried-Blackmore AC, McEwen BS, Bulloch K (2007) Microglia derived from aging mice exhibit an altered inflammatory profile. Glia 55:412–424. https://doi.org/10.1002/glia.20468
Sixt M, Engelhardt B, Pausch F., Hallmann R, Wendler O, Sorokin LM (2001) Endothelial cell laminin isoforms, laminins 8 and 10, play decisive roles in T cell recruitment across the blood–brain barrier in experimental autoimmune encephalomyelitis. J Cell Biol 153(5):933–946
Streit WJ, Braak H, Xue Q-S, Bechmann I (2009) Dystrophic (senescent) rather than activated microglial cells are associated with tau pathology and likely precede neurodegeneration in Alzheimer’s disease. Acta Neuropathol 118:475–485. https://doi.org/10.1007/s00401-009-0556-6
Su EJ, Cao C, Fredriksson L, Nilsson I, Stefanitsch C, Stevenson TK, Zhao J, Ragsdale M, Sun Y-Y, Yepes M, Kuan C-Y, Eriksson U, Strickland DK, Lawrence DA, Zhang L (2017) Microglial-mediated PDGF-CC activation increases cerebrovascular permeability during ischemic stroke. Acta Neuropathol 134:585–604. https://doi.org/10.1007/s00401-017-1749-z
Sweeney MD, Sagare AP, Zlokovic BV (2018) Blood–brain barrier breakdown in Alzheimer disease and other neurodegenerative disorders. Nat Rev Neurol 14:133. https://doi.org/10.1038/nrneurol.2017.188
Tay TL, Mai D, Dautzenberg J, Fernández-Klett F, Lin G, Datta M, Drougard A, Stempfl T, Ardura-Fabregat A, Staszewski O, Margineanu A, Sporbert A, Steinmetz LM, Pospisilik JA, Jung S, Priller J, Grün D, Ronneberger O, Prinz M (2017) A new fate mapping system reveals context-dependent random or clonal expansion of microglia. Nat Neurosci 20:793–803. https://doi.org/10.1038/nn.4547
Thomsen MS, Routhe LJ, Moos T (2017) The vascular basement membrane in the healthy and pathological brain. J Cereb Blood Flow Metab 37:3300–3317. https://doi.org/10.1177/0271678X17722436
Tischer J, Krueger M, Mueller W, Staszewski O, Prinz M, Streit WJ, Bechmann I (2016) Inhomogeneous distribution of Iba-1 characterizes microglial pathology in Alzheimer’s disease. Glia 64:1562–1572. https://doi.org/10.1002/glia.23024
Toft-Hansen H, Nuttall RK, Edwards DR, Owens T (2004) Key metalloproteinases are expressed by specific cell types in experimental autoimmune encephalomyelitis. J Immunol 173:5209–5218. https://doi.org/10.4049/jimmunol.173.8.5209
Tremblay M-È, Zettel ML, Ison JR, Allen PD, Majewska AK (2012) Effects of aging and sensory loss on glial cells in mouse visual and auditory cortices. Glia 60:541–558. https://doi.org/10.1002/glia.22287
Wieghofer P, Knobeloch K-P, Prinz M (2015) Genetic targeting of microglia. Glia 63:1–22. https://doi.org/10.1002/glia.22727
Wolf SA, Boddeke HWGM, Kettenmann H (2017) Microglia in Physiology and Disease. Annu Rev Physiol 79:619–643. https://doi.org/10.1146/annurev-physiol-022516-034406
Wu C, Ivars F, Anderson P, Hallmann R, Vestweber D, Nilsson P, Robenek H, Tryggvason K, Song J, Korpos E, Loser K, Beissert S, Georges-Labouesse E, Sorokin LM (2009) Endothelial basement membrane laminin alpha5 selectively inhibits T lymphocyte extravasation into the brain. Nat Med 15:519–527. https://doi.org/10.1038/nm.1957
Yousif LF, Di Russo J, Sorokin L (2013) Laminin isoforms in endothelial and perivascular basement membranes. Cell Adh Migr 7:101–110. https://doi.org/10.4161/cam.22680
Zhao Z, Nelson AR, Betsholtz C, Zlokovic BV (2015) Establishment and dysfunction of the blood–brain barrier. Cell 163:1064–1078. https://doi.org/10.1016/j.cell.2015.10.067
Funding
This work was supported by Deutsche Forschungsgemeinschaft (SFB Grant 1052 ‘Obesity mechanisms’) to IB.
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflicts of interest
The authors declare that there is no conflict of interest.
Ethical approval
All procedures performed in studies involving human tissue were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.
Research involving human and/or animal participants
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Ingo Bechmann and Martin Krueger these authors equally contributed
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Joost, E., Jordão, M.J.C., Mages, B. et al. Microglia contribute to the glia limitans around arteries, capillaries and veins under physiological conditions, in a model of neuroinflammation and in human brain tissue. Brain Struct Funct 224, 1301–1314 (2019). https://doi.org/10.1007/s00429-019-01834-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00429-019-01834-8