Journal Mobile Options
Table of Contents
Vol. 94, No. 4, 2008
Issue release date: November 2008
Neonatology 2008;94:255–266

Pretreatment with Low Doses of Erythropoietin Ameliorates Brain Damage in Periventricular Leukomalacia by Targeting Late Oligodendrocyte Progenitors: A Rat Model

Mizuno K. · Hida H. · Masuda T. · Nishino H. · Togari H.
Departments of aPediatrics and Neonatology and bNeurophysiology and Brain Science, Nagoya City University Graduate School of Medical Science, Nagoya, Japan

Individual Users: Register with Karger Login Information

Please create your User ID & Password

Contact Information

I have read the Karger Terms and Conditions and agree.

To view the fulltext, please log in

To view the pdf, please log in


Background: One of the pathological hallmarks of periventricular leukomalacia (PVL) is the selective vulnerability of late oligodendrocyte progenitors (preoligodendrocytes; preOLs) to hypoxia-ischemia (H-I). It is unknown whether recombinant human erythropoietin (rhEPO) protects preOLs in vivo. Objectives: To develop a rat PVL model in which preOLs are selectively damaged and exhibit similar pathological changes to diffuse-type human PVL, various conditions of H-I were compared in P2–P7 rats (P2 = postnatal day 2). To evaluate the effect of rhEPO on oligoprotection (preOLs), rhEPO was administered to P3 PVL rats. Methods: After counts of NG2-positive and O4-positive cells were performed in P2–P7 rats, right common carotid artery occlusion followed by 6% O2 for 0–120 min was performed in P2–P4 rats. The mortality and histological alterations after hematoxylin/eosin staining and ED1 immunostaining were assessed 2 days after H-I. Various doses of rhEPO (1–30,000 U/kg i.p.) were administered to PVL rats 15 min before administration of 6% O2. Results: Double-positive cells for NG2 and O4 were detected from P2, and their number gradually increased until P7. Although right common carotid artery occlusion with 6% O2 for 60 min resulted in a relatively high proportion of deaths in P2–P4 rats, typical histological changes in the PVL diffuse component were found in most surviving P3 animals. With 50–100 U/kg rhEPO, the histological damage was attenuated. Conclusions: Histological changes similar to those seen in the PVL diffuse component were induced by H-I in P3 rats, in which preOLs were gradually developing, and a low dose of rhEPO was effective in the treatment of brain damage induced by H-I.

Copyright / Drug Dosage

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.


  1. Wood NS, Marlow N, Costeloe K, Gibson AT, Wilkinson AR: Neurologic and developmental disability after extremely preterm birth. N Engl J Med 2000;343:378–384.
  2. Back SA, Rivkees SA: Emerging concepts in periventricular white matter injury. Semin Perinatol 2004;28:405–414.
  3. Volpe JJ: Neurobiology of periventricular leukomalacia in the premature infant. Pediatr Res 2001;50:553–562.
  4. Fujimoto S, Togari H, Takashima S, Funato M, Yoshioka H, Ibara S, Tatsuno M: National survey of periventricular leukomalacia in Japan. Acta Paediatr Jpn 1998;40:239–243.
  5. Takashima S, Tanaka K: Development of cerebrovascular architecture and its relationship to periventricular leukomalacia. Arch Neurol 1978;35:11–16.
  6. Back SA, Gan X, Li Y, Rosenberg PA, Volpe JJ: Maturation-dependent vulnerability of oligodendrocytes to oxidative stress-induced death caused by glutathione depletion. J Neurosci 1998;18:6241–6253.
  7. Back SA, Luo NL, Borenstein NS, Levine JM, Volpe JJ, Kinney HC: Late oligodendrocyte progenitors coincide with the developmental window of vulnerability for human perinatal white matter injury. J Neurosci 2001;21:1302–1312.
  8. Ballabh P, Xu H, Hu F, Braun A, Smith K, Rivera A, Lou N, Ungvari Z, Goldman SA, Csiszar A, Nedergaard M: Angiogenic inhibition reduces germinal matrix hemorrhage. Nat Med 2007;13:477–485.
  9. Back SA, Han BH, Luo NL, Chricton CA, Xanthoudakis S, Tam J, Arvin KL, Holtzman DM: Selective vulnerability of late oligodendrocyte progenitors to hypoxia-ischemia. J Neurosci 2002;22:455–463.
  10. Fern R, Möller T: Rapid ischemic cell death in immature oligodendrocytes: a fatal glutamate release feedback loop. J Neurosci 2000;20:34–42.
  11. Jensen FE: Role of glutamate receptors in periventricular leukomalacia. J Child Neurol 2005;20:950–959.
  12. Hagberg H, Peebles D, Mallard C: Models of white matter injury: comparison of infectious, hypoxic-ischemic, and excitotoxic insults. Ment Retard Dev Disabil Res Rev 2002;8:30–38.
  13. Bahcekapili N, Uzüm G, Gökkusu C, Kuru A, Ziylan YZ: The relationship between erythropoietin pretreatment with blood-brain barrier and lipid peroxidation after ischemia/reperfusion in rats. Life Sci 2007;80:1245–1251.
  14. Baud O, Green AE, Li J, Wang H, Volpe JJ, Rosenberg PA: Glutathione peroxidase-catalase cooperativity is required for resistance to hydrogen peroxide by mature rat oligodendrocytes. J Neurosci 2004;24:1531–1540.
  15. Mitsufuji N, Yoshioka H, Okano S, Nishiki T, Sawada T: A new model of transient cerebral ischemia in neonatal rats. J Cereb Blood Flow Metab 1996;16:237–243.
  16. Sheldon RA, Chuai J, Ferriero DM: A rat model for hypoxic-ischemic brain damage in very premature infants. Biol Neonate 1996;69:327–341.
  17. Ota A, Ikeda T, Ikenoue T, Toshimori K: Sequence of neuronal responses assessed by immunohistochemistry in the newborn rat brain after hypoxia-ischemia. Am J Obstet Gynecol 1997;177:519–526.
  18. Vannucci RC, Rossini A, Towfighi J, Vannucci SJ: Measuring the accentuation of the brain damage that arises from perinatal cerebral hypoxia-ischemia. Biol Neonate 1997;72:187–191.
  19. Uehara H, Yoshioka H, Kawase S, Nagai H, Ohmae T, Hasegawa K, Sawada T: A new model of white matter injury in neonatal rats with bilateral carotid artery occlusion. Brain Res 1999;837:213–220.
  20. Derugin N, Wendland M, Muramatsu K, Roberts TP, Gregory G, Ferriero DM, Vexler ZS: Evolution of brain injury after transient middle cerebral artery occlusion in neonatal rats. Stroke 2000;31:1752–1761.
  21. Oakden E, Chiswick M, Rothwell N, Loddick S: The influence of litter size on brain damage caused by hypoxic-ischemic injury in the neonatal rat. Pediatr Res 2002;52:692–696.
  22. Sheldon RA, Sedik C, Ferriero DM: Strain-related brain injury in neonatal mice subjected to hypoxia-ischemia. Brain Res 1998;810:114–122.
  23. Skoff RP, Bessert DA, Barks JD, Song D, Cerghet M, Silverstein FS: Hypoxic-ischemic injury results in acute disruption of myelin gene expression and death of oligodendroglial precursors in neonatal mice. Int J Dev Neurosci 2001;19:197–208.
  24. Yoshioka H, Goma H, Nioka S, Ochi M, Miyake H, Zaman A, Masumura M, Sawada T, Chance B: Bilateral carotid artery occlusion causes periventricular leukomalacia in neonatal dogs. Brain Res Dev Brain Res 1994;78:273–278.
  25. Ohyu J, Marumo G, Ozawa H, Takashima S, Nakajima K, Kohsaka S, Hamai Y, Machida Y, Kobayashi K, Ryo E, Baba K, Kozuma S, Okai T, Taketani Y: Early axonal and glial pathology in fetal sheep brains with leukomalacia induced by repeated umbilical cord occlusion. Brain Dev 1999;21:248–252.
  26. Craig A, Ling Luo N, Beardsley DJ, Wingate-Pearse N, Walker DW, Hohimer AR, Back SA: Quantitative analysis of perinatal rodent oligodendrocyte lineage progression and its correlation with human. Exp Neurol 2003;181:231–240.
  27. Sakanaka M, Wen TC, Matsuda S, Masuda S, Morishita E, Nagao M, Sasaki R: In vivo evidence that erythropoietin protects neurons from ischemic damage. Proc Natl Acad Sci USA 1988;95:4635–4640.
  28. Kawakami M, Sekiguchi M, Sato K, Kozaki S, Takahashi M: Erythropoietin receptor-mediated inhibition of exocytotic glutamate release confers neuroprotection during chemical ischemia. J Biol Chem 2001;276:39469–39475.
  29. Kumral A, Gonenc S, Acikgoz O, Sonmez A, Genc K, Yilmaz O, Gokmen N, Duman N, Ozkan H: Erythropoietin increases glutathione peroxidase enzyme activity and decreases lipid peroxidation levels in hypoxic-ischemic brain injury in neonatal rats. Biol Neonate 2005;87:15–18.
  30. Sugawa M, Sakurai Y, Ishikawa-Ieda Y, Suzuki H, Asou H: Effects of erythropoietin on glial cell development: oligodendrocyte maturation and astrocyte proliferation. Neurosci Res 2002;44:391–403.
  31. Hida H, Takeda M, Soliven B: Ceramide inhibits inwardly rectifying K+ currents via a Ras- and Raf-1-dependent pathway in cultured oligodendrocytes. J Neurosci 1998;18:8712–8719.
  32. Yang Z, Watanabe M, Nishiyama A: Optimization of oligodendrocyte progenitor cell culture method for enhanced survival. J Neurosci Methods 2005;149:50–56.
  33. Demers EJ, McPherson RJ, Juul SE: Erythropoietin protects dopaminergic neurons and improves neurobehavioral outcome in juvenile rats after neonatal hypoxia-ischemia. Pediatr Res 2005;58:297–301.
  34. Back SA, Luo NL, Borenstein NS, Volpe JJ, Kinney HC: Arrested oligodendrocyte lineage progression during human cerebral white matter development: dissociation between the timing of progenitor differentiation and myelinogenesis. J Neuropathol Exp Neurol 2002;61:197–211.
  35. Riddle A, Luo NL, Manese M, Beardsley DJ, Green L, Rorvik D, Kelly KA, Barlow CH, Kelly JJ, Hosimer AR, Back SA: Spatial heterogeneity in oligodendrocyte lineage maturation and not cerebral blood flow predicts fetal ovine periventricular white matter injury. J Neurosci 2006;26:3045–3055.
  36. Wu H, Ren B, Zhu J, Dong G, Xu B, Wang C, Zheng X, Jing H: Pretreatment with recombinant human erythropoietin attenuates ischemia-reperfusion-induced lung injury in rats. Eur J Cardiothorac Surg 2006;29:902–907.
  37. Vesy DA, Cheung C, Pat B, Endre Z, Gobe G, Johnson DW: Erythropoietin protects against ischemic acute renal injury. Nephrol Dial Transplant 2004;19:348–355.
  38. Fragoso G, Martínez-Bermúdez AK, Liu HN, Khorchid A, Chemtob S, Mushynski WE, Almazan G: Developmental differences in H2O2-induced oligodendrocyte cell death: role of glutathione, mitogen-activated protein kinases and caspase 3. J Neurochem 2004;90:392–404.
  39. Sánchez-Gómez MV, Alberdi E, Ibarretxe G, Torre I, Matute C: Caspase-dependent and caspase-independent oligodendrocyte death mediated by AMPA and kainate receptors. J Neurosci 2003;23:9519–9528.
  40. Leist M, Ghezzi P, Grasso G, Bianchi R, Villa P, Fratelli M, Savino C, Bianchi M, Nielsen J, Gerwien J, Kallunki P, Larsen AK, Helboe L, Christensen S, Pedersen LO, Nielsen M, Torup L, Sager T, Sfacteria A, Erbayraktar S, Erbayraktar Z, Gokmen N, Yilmaz O, Cerami-Hand C, Xie QW, Coleman T, Cerami A, Brines M: Derivatives of erythropoietin that are tissue protective but not erythropoietic. Science 2004;305:239–242.
  41. Wang X, Zhu C, Wang X, Gerwien J, Schrattenholz A, Sandberg M, Leist M, Blomgren K: The nonerythropoietic asialoerythropoietin protects against neonatal hypoxia-ischemia as potently as erythropoietin. J Neurochem 2004;91:900–910.
  42. Sun Y, Calvert JW, Zhang JH: Neonatal hypoxia/ischemia is associated with decreased inflammatory mediators after erythropoietin administration. Stroke 2005;36:1672–1678.
  43. Kellert BA, McPherson RJ, Juul SE: A comparison of high-dose recombinant erythropoietin treatment regimens in brain-injured neonatal rats. Pediatr Res 2007;61:451–455.
  44. Statler PA, McPherson RJ, Bauer LA, Kellert BA, Juul SE: Pharmacokinetics of high-dose recombinant erythropoietin in plasma and brain of neonatal rats. Pediatr Res 2007;61:671–675.
  45. Sun Y, Zhou C, Polk P, Nanda A, Zhang JH: Mechanisms of erythropoietin-induced brain protection in neonatal hypoxia-ischemia rat model. J Cereb Blood Flow Metab 2004;24:259–270.
  46. Itoh T, Beesley J, Itoh A, Cohen AS, Kavanaugh B, Coulter DA, Grinspan JB, Pleasure D: AMPA glutamate receptor-mediated calcium signaling is transiently enhanced during development of oligodendrocytes. J Neurochem 2002;81:390–402.
  47. Follett PL, Deng W, Dai W, Talos DM, Massillon LJ, Rosenberg PA, Volpe JJ, Jensen FE: Glutamate receptor-mediated oligodendrocyte toxicity in periventricular leukomalacia: a protective role for topiramate. J Neurosci 2004;24:4412–4420.
  48. Káradóttir R, Cavelier P, Bergersen LH, Attwell D: NMDA receptors are expressed in oligodendrocytes and activated in ischaemia. Nature 2005;438:1162–1166.
  49. Salter MG, Fern R: NMDA receptors are expressed in developing oligodendrocyte process and mediate injury. Nature 2005;438:1167–1171.
  50. Hirrlinger J, Resch A, Gutterer JM, Dringen R: Oligodendroglial cells in culture effectively dispose of exogenous hydrogen peroxide: comparison with cultured neurones, astroglial and microglial cells. J Neurochem 2002;82:635–644.
  51. Kress G, Dineley KE, Reynolds IJ: The relationship between intracellular free iron and cell injury in cultured neurons, astrocytes, and oligodendrocytes. J Neurosci 2002;22:5848–5855.
  52. Palmer C, Menzies SL, Roberts RL, Pavlick G, Connor JR: Changes in iron histochemistry after hypoxic-ischemic brain injury in the neonatal rat. J Neurosci Res 1999;56:60–71.
  53. Brines M, Ghezzi P, Keenan S, Agnello D, de Lanerolle NC, Cerami C, Itri LM, Cerami A: Erythropoietin crosses the blood-brain barrier to protect against experimental brain injury. Proc Natl Acad Sci USA 2000;97:10526–10531.
  54. Cai Z, Lin S, Fan LW, Pang Y, Rhodes PG: Minocycline alleviates hypoxic-ischemic injury to developing oligodendrocytes in the neonatal rat brain. Neuroscience 2006;137:425–435.

Pay-per-View Options
Direct payment This item at the regular price: USD 38.00
Payment from account With a Karger Pay-per-View account (down payment USD 150) you profit from a special rate for this and other single items.
This item at the discounted price: USD 26.50