Objective: To investigate the effects of neonatal stroke on progenitor cells lining the lateral ventricles. Methods: Intraventricular injection of replication-incompetent green fluorescent protein (GFP)-expressing lentivirus was performed in postnatal day 1 (P1) rats to specifically label radial glia/type B neural stem cells and ependymal cells of the lateral ventricle. A subset of animals was exposed to transient middle cerebral artery occlusion (MCAO) at P7, with mild or moderate injury confirmed by diffusion-weighted MRI and histology. Newborn cells were identified by GFP expression, location and expression of cell type-specific markers in the striatum, cortex and olfactory bulb using confocal microscopy and systematic random sampling. Results: Three weeks lentiviral GFP transduction of cells in the lateral ventricle, abundant GFP-expressing neurons and glia were identified in the rostral migratory stream, olfactory bulb and striatum as expected from labeling the subventricular zone (SVZ) type B neural stem cell lineage. Two weeks following mild or severe focal stroke at P7, no GFP-expressing neurons were detected in striatum or cortex although some single-labeled doublecortin+ immature neurons were detected in the penumbra. The densities of GFP+/ glial fibrillary acidic protein (GFAP)+ astrocytes and GFP+/O4+ oligodendrocytes were reduced in the striatum following MCAO (4.8 ± 1.02 vs. 2.5 ± 0.4 cells/high-power field, HPF; p = 0.005; 2.8± 1 vs. 0.5 ± 0.2 cells/HPF, p = 0.008). Furthermore, there was a reduction of GFP+ cells in the olfactory bulb following MCAO (58.8 ± 14.9 vs. 19.6 ± 5.4 cells/HPF, p = 0.025). Finally, there was an increased percentage of GFP+/GFAP+ cells (70 vs. 50%), with a decreased proportion of GFP+/O4+ cells (14 vs. 30%) in injured animals. Conclusion: Neurogenesis originating from cells of the lateral ventricle, including SVZ type B cells, is significantly reduced following neonatal stroke. Furthermore, neonatal stroke disrupts gliogenesis in the striatum, decreasing overall numbers of new glia and shifting the population towards astrocytes.
© 2010 S. Karger AG, Basel
- Stem cells
- Arvidsson A, Collin T, Kirik D, Kokaia Z, Lindvall O: Neuronal replacement from endogenous precursors in the adult brain after stroke. Nat Med 2002;8:963–970.
- Parent JM, Vexler ZS, Gong C, Derugin N, Ferriero DM: Rat forebrain neurogenesis and striatal neuron replacement after focal stroke. Ann Neurol 2002;52:802–813.
- Yang Z, Levison SW: Hypoxia/ischemia expands the regenerative capacity of progenitors in the perinatal subventricular zone. Neuroscience 2006;139:555–564.
- Plane JM, Liu R, Wang TW, Silverstein FS, Parent JM: Neonatal hypoxic-ischemic injury increases forebrain subventricular zone neurogenesis in the mouse. Neurobiol Dis 2004;16:585–595.
- Ong J, Plane JM, Parent JM, Silverstein FS: Hypoxic-ischemic injury stimulates subventricular zone proliferation and neurogenesis in the neonatal rat. Pediatr Res 2005;58:600–606.
- Ackman JB, Siddiqi F, Walikonis RS, LoTurco JJ: Fusion of microglia with pyramidal neurons after retroviral infection. J Neurosci 2006;26:11413–11422.
- Kuhn HG, Cooper-Kuhn CM: Bromodeoxyuridine and the detection of neurogenesis. Curr Pharm Biotechnol 2007;8:127–131.
- Taupin P: BrdU immunohistochemistry for studying adult neurogenesis: Paradigms, pitfalls, limitations, and validation. Brain Res Rev 2007;53:198–214.
- Sergent-Tanguy S, Michel DC, Neveu I, Naveilhan P: Long-lasting coexpression of nestin and glial fibrillary acidic protein in primary cultures of astroglial cells with a major participation of nestin(+)/GFAP(–) cells in cell proliferation. J Neurosci Res 2006;83:1515–1524.
- Lois C, Alvarez-Buylla A: Proliferating subventricular zone cells in the adult mammalian forebrain can differentiate into neurons and glia. Proc Natl Acad Sci USA 1993;90:2074–2077.
- Doetsch F, Alvarez-Buylla A: Network of tangential pathways for neuronal migration in adult mammalian brain. Proc Natl Acad Sci USA 1996;93:14895–14900.
- Levison SW, Goldman JE: Both oligodendrocytes and astrocytes develop from progenitors in the subventricular zone of postnatal rat forebrain. Neuron 1993;10:201–212.
- Suzuki SO, Goldman JE: Multiple cell populations in the early postnatal subventricular zone take distinct migratory pathways: a dynamic study of glial and neuronal progenitor migration. J Neurosci 2003;23:4240–4250.
- Doetsch F, Caille I, Lim DA, Garcia-Verdugo JM, Alvarez-Buylla A: Subventricular zone astrocytes are neural stem cells in the adult mammalian brain. Cell 1999;97:703–716.
- Merkle FT, Tramontin AD, Garcia-Verdugo JM, Alvarez-Buylla A: Radial glia give rise to adult neural stem cells in the subventricular zone. Proc Natl Acad Sci USA 2004;101:17528–17532.
- Mirzadeh Z, Merkle FT, Soriano-Navarro M, Garcia-Verdugo JM, Alvarez-Buylla A: Neural stem cells confer unique pinwheel architecture to the ventricular surface in neurogenic regions of the adult brain. Cell Stem Cell 2008;3:265–278.
- Merkle FT, Alvarez-Buylla A: Neural stem cells in mammalian development. Curr Opin Cell Biol 2006;18:704–709.
- Lois C, Hong EJ, Pease S, Brown EJ, Baltimore D: Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors. Science 2002;295:868–872.
- Geraerts M, Eggermont K, Hernandez-Acosta P, Garcia-Verdugo JM, Baekelandt V, Debyser Z: Lentiviral vectors mediate efficient and stable gene transfer in adult neural stem cells in vivo. Hum Gene Ther 2006;17:635–650.
- Carlen M, Meletis K, Goritz C, Darsalia V, Evergren E, Tanigaki K, Amendola M, Barnabe-Heider F, Yeung MS, Naldini L, Honjo T, Kokaia Z, Shupliakov O, Cassidy RM, Lindvall O, Frisen J: Forebrain ependymal cells are notch-dependent and generate neuroblasts and astrocytes after stroke. Nat Neurosci 2009;12:259–267.
- Derugin N, Ferriero DM, Vexler ZS: Neonatal reversible focal cerebral ischemia: a new model. Neurosci Res 1998;32:349–353.
- Gonzalez FF, McQuillen P, Mu D, Chang Y, Wendland M, Vexler Z, Ferriero DM: Erythropoietin enhances long-term neuroprotection and neurogenesis in neonatal stroke. Dev Neurosci 2007;29:321–330.
- Yang Z, Covey MV, Bitel CL, Ni L, Jonakait GM, Levison SW: Sustained neocortical neurogenesis after neonatal hypoxic/ischemic injury. Ann Neurol 2007;61:199–208.
- Zerlin M, Levison SW, Goldman JE: Early patterns of migration, morphogenesis, and intermediate filament expression of subventricular zone cells in the postnatal rat forebrain. J Neurosci 1995;15:7238–7249.
- Chang YS, Mu D, Wendland M, Sheldon RA, Vexler ZS, McQuillen PS, Ferriero DM: Erythropoietin improves functional and histological outcome in neonatal stroke. Pediatr Res 2005;58:106–111.
- Burns KA, Ayoub AE, Breunig JJ, Adhami F, Weng WL, Colbert MC, Rakic P, Kuan CY: Nestin-creer mice reveal DNA synthesis by nonapoptotic neurons following cerebral ischemia hypoxia. Cereb Cortex 2007;17:2585–2592.
- Levison SW, Rothstein RP, Romanko MJ, Snyder MJ, Meyers RL, Vannucci SJ: Hypoxia/ischemia depletes the rat perinatal subventricular zone of oligodendrocyte progenitors and neural stem cells. Dev Neurosci 2001;23:234–247.
- Romanko MJ, Zhu C, Bahr BA, Blomgren K, Levison SW: Death effector activation in the subventricular zone subsequent to perinatal hypoxia/ischemia. J Neurochem 2007;103:1121–1131.
- Iwai M, Ikeda T, Hayashi T, Sato K, Nagata T, Nagano I, Shoji M, Ikenoue T, Abe K: Temporal profile of neural stem cell proliferation in the subventricular zone after ischemia/hypoxia in the neonatal rat brain. Neurol Res 2006;28:461–468.
- Kadam SD, Mulholland JD, McDonald JW, Comi AM: Poststroke subgranular and rostral subventricular zone proliferation in a mouse model of neonatal stroke. J Neurosci Res 2009;87:2653–2666.
- Cooper-Kuhn CM, Kuhn HG: Is it all DNA repair? Methodological considerations for detecting neurogenesis in the adult brain. Brain Res Dev Brain Res 2002;134:13–21.
- Kuan CY, Schloemer AJ, Lu A, Burns KA, Weng WL, Williams MT, Strauss KI, Vorhees CV, Flavell RA, Davis RJ, Sharp FR, Rakic P: Hypoxia-ischemia induces DNA synthesis without cell proliferation in dying neurons in adult rodent brain. J Neurosci 2004;24:10763–10772.
- Iwai M, Cao G, Yin W, Stetler RA, Liu J, Chen J: Erythropoietin promotes neuronal replacement through revascularization and neurogenesis after neonatal hypoxia/ischemia in rats. Stroke 2007;38:2795–2803.
- Chen ZY, Asavaritikrai P, Prchal JT, Noguchi CT: Endogenous erythropoietin signaling is required for normal neural progenitor cell proliferation. J Biol Chem 2007;282:25875–25883.
- Seri B, Herrera DG, Gritti A, Ferron S, Collado L, Vescovi A, Garcia-Verdugo JM, Alvarez-Buylla A: Composition and organization of the SCZ: a large germinal layer containing neural stem cells in the adult mammalian brain. Cereb Cortex 2006;16 (suppl 1):I103–I111.
- Laskowski A, Howell OW, Sosunov AA, McKhann G, Gray WP: NPY mediates basal and seizure-induced proliferation in the subcallosal zone. Neuroreport 2007;18:1005–1008.
- 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: Isolation and characterization of neural stem/progenitor cells from post-stroke cerebral cortex in mice. Eur J Neurosci 2009;29:1842–1852.
- Burns KA, Murphy B, Danzer SC, Kuan CY: Developmental and post-injury cortical gliogenesis: a genetic fate-mapping study with nestin-creER mice. Glia 2009;57:1115–1129.
- Biran V, Joly LM, Heron A, Vernet A, Vega C, Mariani J, Renolleau S, Charriaut-Marlangue C: Glial activation in white matter following ischemia in the neonatal P7 rat brain. Exp Neurol 2006;199:103–112.
- Segovia KN, McClure M, Moravec M, Luo NL, Wan Y, Gong X, Riddle A, Craig A, Struve J, Sherman LS, Back SA: Arrested oligodendrocyte lineage maturation in chronic perinatal white matter injury. Ann Neurol 2008;63:520–530.
- Sellers DL, Maris DO, Horner PJ: Postinjury niches induce temporal shifts in progenitor fates to direct lesion repair after spinal cord injury. J Neurosci 2009;29:6722–6733.
- Guo F, Ma J, McCauley E, Bannerman P, Pleasure D: Early postnatal proteolipid promoter-expressing progenitors produce multilineage cells in vivo. J Neurosci 2009;29:7256–7270.
- Rothstein RP, Levison SW: Gray matter oligodendrocyte progenitors and neurons die caspase-3-mediated deaths subsequent to mild perinatal hypoxic/ischemic insults. Dev Neurosci 2005;27:149–159.
Prof. Donna M. Ferriero, MD
University of California, San Francisco
521 Parnassus Avenue, Rm C215
San Francisco, CA 94143-0663 (USA)
Tel. +1 415 502 5820, Fax +1 415 502 5821, E-Mail email@example.com
Received: July 27, 2009
Accepted after revision: January 21, 2010
Published online: May 4, 2010
Number of Print Pages : 13
Number of Figures : 5, Number of Tables : 0, Number of References : 43
Additional supplementary material is available online - Number of Parts : 1
Vol. 32, No. 2, Year 2010 (Cover Date: July 2010)
Journal Editor: Levison S.W. (Newark, N.J.)
ISSN: 0378-5866 (Print), eISSN: 1421-9859 (Online)
For additional information: http://www.karger.com/DNE
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.