Abstract
The present study aimed to provide a detailed characterization of the cellular phenotypes of nestin-positive cells in a rat model of ischemic stroke. Nestin-positive cells included reactive astrocytes in the peri-infarct region. In the ischemic core, in which astrocytes had virtually disappeared, nestin expression was exclusively associated with the vasculature, including the microvasculature and larger caliber vessels. Induction of nestin expression in the ischemic core occurred by 3 days post-ischemia. Nestin expression continued through at least 28 days post-ischemia but the cellular profiles of nestin-positive cells changed over this period. In the ischemic core at day 3, nestin-positive cells frequently had long processes that ran parallel along the longitudinal axis of the vasculature. These cells were highly proliferative and expressed the transcription factor for neural/glial progenitors, Sox9. Based on their morphological characteristics and on a double-labeling study, most nestin-positive cells were clearly distinguishable from vasculature-associated cells including endothelial cells, smooth muscle cells and microglia/macrophages. Immunoelectron microscopic findings demonstrated that most nestin-positive cells lay in the perivascular space and had macrophage-like features, indicating morphological similarity to perivascular macrophages. Nestin expression was still associated with the vasculature 14 days after ischemia but appeared in fibroblast-like cells. Thus, our data indicated that, in the ischemic core, nestin expression was not limited to a progenitor/stem cell population but was induced in the vasculature-associated cells. These cell types included perivascular macrophages and fibroblast-like cells that appeared to undergo dynamic structural changes. These results suggest that nestin facilitates cellular structural remodeling in response to ischemic injury.
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Alliot F, Rutin J, Leenen PJ, Pessac B (1999) Pericytes and periendothelial cells of brain parenchyma vessels co-express aminopeptidase N, aminopeptidase A, and nestin. J Neurosci Res 58:367–378
Audoy-Remus J, Richard JF, Soulet D, Zhou H, Kubes P, Vallieres L (2008) Rod-shaped monocytes patrol the brain vasculature and give rise to perivascular macrophages under the influence of proinflammatory cytokines and angiopoietin-2. J Neurosci 28:10187–10199
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 68:242–249
Bechmann I, Galea I, Perry VH (2007) What is the blood–brain barrier (not)? Trends Immunol 28:5–11
Brierley JB, Brown AW (1982) The origin of lipid phagocytes in the central nervous system. II. The adventitia of blood vessels. J Comp Neurol 211:407–417
Burggraf D, Trinkl A, Burk J, Martens HK, Dichgans M, Hamann GF (2008) Vascular integrin immunoreactivity is selectively lost on capillaries during rat focal cerebral ischemia and reperfusion. Brain Res 1189:189–197
Calderone A (2012) Nestin+ cells and healing the infarcted heart. Am J Physiol Heart Circ Physiol 302:H1–H9
Chen J, Boyle S, Zhao M, Su W, Takahashi K, Davis L, Decaestecker M, Takahashi T, Breyer MD, Hao CM (2006) Differential expression of the intermediate filament protein nestin during renal development and its localization in adult podocytes. J Am Soc Nephrol 17:1283–1291
Chu MS, Chang CF, Yang CC, Bau YC, Ho LL, Hung SC (2006) Signalling pathway in the induction of neurite outgrowth in human mesenchymal stem cells. Cell Signal 18:519–530
Chu X, Fu X, Zou L, Qi C, Li Z, Rao Y, Ma K (2007) Oncosis, the possible cell death pathway in astrocytes after focal cerebral ischemia. Brain Res 1149:157–164
Dahlstrand J, Collins VP, Lendahl U (1992) Expression of the class VI intermediate filament nestin in human central nervous system tumors. Cancer Res 52:5334–5341
del Zoppo GJ, Mabuchi T (2003) Cerebral microvessel responses to focal ischemia. J Cereb Blood Flow Metab 23:879–894
Domowicz MS, Henry JG, Wadlington N, Navarro A, Kraig RP, Schwartz NB (2011) Astrocyte precursor response to embryonic brain injury. Brain Res 1389:35–49
Dore-Duffy P (2003) Isolation and characterization of cerebral microvascular pericytes. Methods Mol Med 89:375–382
Graeber MB, Streit WJ (1990) Perivascular microglia defined. Trends Neurosci 13:366
Graeber MB, Streit WJ, Kreutzberg GW (1989) Identity of ED2-positive perivascular cells in rat brain. J Neurosci Res 22:103–106
Graeber MB, Streit WJ, Büringer D, Sparks DL, Kreutzberg GW (1992) Ultrastructural location of major histocompatibility complex (MHC) class II positive perivascular cells in histologically normal human brain. J Neuropathol Exp Neurol 51:303–311
Hamann GF, Okada Y, Fitridge R, del Zoppo GJ (1995) Microvascular basal lamina antigens disappear during cerebral ischemia and reperfusion. Stroke 26:2120–2126
Herrmann H, Aebi U (2000) Intermediate filaments and their associates: multi-talented structural elements specifying cytoarchitecture and cytodynamics. Curr Opin Cell Biol 12:79–90
Hickey WF, Kimura H (1988) Perivascular microglial cells of the CNS are bone marrow-derived and present antigen in vivo. Science 239:290–292
Hockfield S, McKay RD (1985) Identification of major cell classes in the developing mammalian nervous system. J Neurosci 5:3310–3328
Jin H, Aiyer A, Su J, Borgstrom P, Stupack D, Friedlander M, Varner J (2006) A homing mechanism for bone marrow-derived progenitor cell recruitment to the neovasculature. J Clin Invest 116:652–662
Kachinsky AM, Dominov JA, Miller JB (1995) Intermediate filaments in cardiac myogenesis: nestin in the developing mouse heart. J Histochem Cytochem 43:843–847
Kida S, Steart PV, Zhang ET, Weller RO (1993) Perivascular cells act as scavengers in the cerebral perivascular spaces and remain distinct from pericytes, microglia and macrophages. Acta Neuropathol 85:646–652
Kim SY, Lee SH, Kim BM, Kim EH, Min BH, Bendayan M, Park IS (2004) Activation of nestin-positive duct stem (NPDS) cells in pancreas upon neogenic motivation and possible cytodifferentiation into insulin-secreting cells from NPDS cells. Dev Dyn 230:1–11
Kishaba Y, Matsubara D, Niki T (2010) Heterogeneous expression of nestin in myofibroblasts of various human tissues. Pathol Int 60:378–385
Lendahl U, Zimmerman LB, McKay RD (1990) CNS stem cells express a new class of intermediate filament protein. Cell 60:585–595
Li Y, Chopp M (1999) Temporal profile of nestin expression after focal cerebral ischemia in adult rat. Brain Res 838:1–10
Li L, Liu F, Welser-Alves JV, McCullough LD, Milner R (2012) Upregulation of fibronectin and the α5β1 and αvβ3 integrins on blood vessels within the cerebral ischemic penumbra. Exp Neurol 233:283–291
Lin RC, Matesic DF, Marvin M, McKay RD, Brüstle O (1995) Re-expression of the intermediate filament nestin in reactive astrocytes. Neurobiol Dis 2:79–85
Liu D, Smith CL, Barone FC, Ellison JA, Lysko PG, Li K, Simpson IA (1999) Astrocytic demise precedes delayed neuronal death in focal ischemic rat brain. Brain Res Mol Brain Res 68:29–41
Longa EL, Weinstein PR, Carlson S, Cummins R (1989) Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke 20:84–89
Mercier F, Kitasako JT, Hatton GI (2002) Anatomy of the brain neurogenic zones revisited: fractones and the fibroblast/macrophage network. J Comp Neurol 451:170–188
Mokrý J, Ehrmann J, Karbanová J, Cízková D, Soukup T, Suchánek J, Filip S, Kolár Z (2008) Expression of intermediate filament nestin in blood vessels of neural and non-neural tissues. Acta Med (Hradec Kralove) 51:173–179
Morshead CM, Reynolds BA, Craig CG, McBurney MW, Staines WA, Morassutti D, Weiss S, van der Kooy D (1994) Neural stem cells in the adult mammalian forebrain: a relatively quiescent subpopulation of subependymal cells. Neuron 13:1071–1082
Nakagomi T, Taguchi A, Fujimori Y, Saino O, Nakano-Doi A, Kubo S, Gotoh A, Soma T, Yoshikawa H, Nishizaki T, Nakagomi N, Stern DM, Matsuyama T (2009) Isolation and characterization of neural stem/progenitor cells from post-stroke cerebral cortex in mice. Eur J Neurosci 29:1842–1852
Nakagomi T, Molnár Z, Nakano-Doi A, Taguchi A, Saino O, Kubo S, Clausen M, Yoshikawa H, Nakagomi N, Matsuyama T (2011) Ischemia-induced neural stem/progenitor cells in the pia mater following cortical infarction. Stem Cells Dev 20:2037–2051
Nakano-Doi A, Nakagomi T, Fujikawa M, Nakagomi N, Kubo S, Lu S, Yoshikawa H, Soma T, Taguchi A, Matsuyama T (2010) Bone marrow mononuclear cells promote proliferation of endogenous neural stem cells through vascular niches after cerebral infarction. Stem Cells 28:1292–1302
Nakayama D, Matsuyama T, Ishibashi-Ueda H, Nakagomi T, Kasahara Y, Hirose H, Kikuchi-Taura A, Stern DM, Mori H, Taguchi A (2010) Injury-induced neural stem/progenitor cells in post-stroke human cerebral cortex. Eur J Neurosci 31:90–98
Ohab JJ, Fleming S, Blesch A, Carmichael ST (2006) A neurovascular niche for neurogenesis after stroke. J Neurosci 26:13007–13016
Oikawa H, Hayashi K, Maesawa C, Masuda T, Sobue K (2010) Expression profiles of nestin in vascular smooth muscle cells in vivo and in vitro. Exp Cell Res 316:940–950
Polfliet MM, Fabriek BO, Daniels WP, Dijkstra CD, van den Berg TK (2006) The rat macrophage scavenger receptor CD163: expression, regulation and role in inflammatory mediator production. Immunobiology 211:419–425
Salazar-Colocho P, Del Río J, Frechilla D (2008) Involvement of the vascular wall in regenerative processes after CA1 ischemic neuronal death. Int J Dev Neurosci 26:541–550
Sejersen T, Lendahl U (1993) Transient expression of the intermediate filament nestin during skeletal muscle development. J Cell Sci 106:1291–1300
Shin HY, Kim JH, Phi JH, Park CK, Kim JE, Kim JH, Paek SH, Wang KC, Kim DG (2008) Endogenous neurogenesis and neovascularization in the neocortex of the rat after focal cerebral ischemia. J Neurosci Res 86:356–367
Shin YJ, Choi JS, Choi JY, Hou Y, Cha JH, Chun MH, Lee MY (2010) Induction of vascular endothelial growth factor receptor-3 mRNA in glial cells following focal cerebral ischemia in rats. J Neuroimmunol 229:81–90
Stolt CC, Lommes P, Sock E, Chaboissier MC, Schedl A, Wegner M (2003) The Sox9 transcription factor determines glial fate choice in the developing spinal cord. Genes Dev 17:1677–1689
Streit WJ, Graeber MB (1993) Heterogeneity of microglial and perivascular cell populations: insights gained from the facial nucleus paradigm. Glia 7:68–74
Takamori Y, Mori T, Wakabayashi T, Nagasaka Y, Matsuzaki T, Yamada H (2009) Nestin positive microglia in adult rat cerebral cortex. Brain Res 1270:10–18
Toma JG, Akhavan M, Fernandes KJ, Barnabe-Heider F, Sadikot A, Kaplan DR, Miller FD (2001) Isolation of multipotent adult stem cells from the dermis of mammalian skin. Nat Cell Biol 3:778–784
Tropepe V, Coles BL, Chiasson BJ, Horsford DJ, Elia AJ, McInnes RR, van der Kooy D (2000) Retinal stem cells in the adult mammalian eye. Science 287:2032–2036
Vallières L, Sawchenko PE (2003) Bone marrow-derived cells that populate the adult mouse brain preserve their hematopoietic identity. J Neurosci 23:5197–5207
Wang N, Stamenovic D (2000) Contribution of intermediate filaments to cell stiffness, stiffening, and growth. Am J Physiol Cell Physiol 279:C188–C194
Wiese C, Rolletschek A, Kania G, Blyszczuk P, Tarasov KV, Tarasova Y, Wersto RP, Boheler KR, Wobus AM (2004) Nestin expression—a property of multi-lineage progenitor cells? Cell Mol Life Sci 61:2510–2522
Williams K, Alvarez X, Lackner AA (2001) Central nervous system perivascular cells are immunoregulatory cells that connect the CNS with the peripheral immune system. Glia 36:156–164
Yamashima T, Tonchev AB, Vachkov IH, Popivanova BK, Seki T, Sawamoto K, Okano H (2004) Vascular adventitia generates neuronal progenitors in the monkey hippocampus after ischemia. Hippocampus 14:861–875
Yamashita T, Ninomiya M, Hernández Acosta P, García-Verdugo JM, Sunabori T, Sakaguchi M, Adachi K, Kojima T, Hirota Y, Kawase T, Araki N, Abe K, Okano H, Sawamoto K (2006) Subventricular zone-derived neuroblasts migrate and differentiate into mature neurons in the post-stroke adult striatum. J Neurosci 26:6627–6636
Yokoyama A, Sakamoto A, Kameda K, Imai Y, Tanaka J (2006) NG2 proteoglycan-expressing microglia as multipotent neural progenitors in normal and pathologic brains. Glia 53:754–768
Zulewski H, Abraham EJ, Gerlach MJ, Daniel PB, Moritz W, Müller B, Vallejo M, Thomas MK, Habener JF (2001) Multipotential nestin-positive stem cells isolated from adult pancreatic islets differentiate ex vivo into pancreatic endocrine, exocrine, and hepatic phenotypes. Diabetes 50:521–533
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This study was supported by the Mid-career Researcher Program through a National Research Foundation of Korea (NRF) grant funded by the MEST (2011–0028319).
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Shin, YJ., Kim, H.L., Park, JM. et al. Characterization of nestin expression and vessel association in the ischemic core following focal cerebral ischemia in rats. Cell Tissue Res 351, 383–395 (2013). https://doi.org/10.1007/s00441-012-1538-x
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DOI: https://doi.org/10.1007/s00441-012-1538-x