Acta Neuropathologica

, Volume 118, Issue 3, pp 391–399 | Cite as

Nitrosative stress and inducible nitric oxide synthase expression in periventricular leukomalacia

  • Robin L. Haynes
  • Rebecca D. Folkerth
  • Felicia L. Trachtenberg
  • Joseph J. Volpe
  • Hannah C. Kinney
Original Paper

Abstract

Periventricular leukomalacia (PVL) is a lesion of the immature cerebral white matter in the perinatal period and associated predominantly with prematurity and cerebral ischemia/reperfusion as well as inflammation due to maternofetal infection. It consists of focal necrosis in the periventricular region and diffuse gliosis with microglial activation and premyelinating oligodendrocyte (pre-OL) injury in the surrounding white matter. We previously showed nitrotyrosine in pre-OLs in PVL, suggesting involvement of nitrosative stress in this disorder. Here we hypothesize that inducible nitric oxide synthase (iNOS) expression is increased in PVL relative to controls. Using immunocytochemistry in human archival tissue, the density of iNOS-expressing cells was determined in the cerebral white matter of 15 PVL cases [29–51 postconceptional (PC) weeks] and 16 control cases (20–144 PC weeks). Using a standardization score of 0–3, the density of iNOS-positive cells was significantly increased in the diffuse component of PVL (score of 1.8 ± 0.3) cases compared to controls (score of 0.7 ± 0.3) (P = 0.01). Intense iNOS expression occurred in reactive astrocytes in acute through chronic stages and in activated microglia primarily in the acute stage, suggesting an early role for microglial iNOS in PVL’s pathogenesis. This study supports an important role for iNOS-induced nitrosative stress in the reactive/inflammatory component of PVL.

Keywords

Nitrotyrosine Oxidative stress Astrocytes Microglia 

Abbreviations

ANCOVA

Analysis of covariance

GA

Gestational

GFAP

Glial fibrillary acidic protein

ICC

Immunocytochemisty

iNOS

Inducible nitric oxide synthase

NO

Nitric oxide

NT

Nitrotyrosine

PC

Postconceptional

PMI

Postmortem interval

PN

Postnatal

pre-OL

Premyelinating oligodendrocyte

PVL

Periventricular leukomalacia

Notes

Acknowledgments

The authors thank Mr. Richard A. Belliveau for assistance in this study. This research was supported by grants from the National Institute of Neurological Diseases and Stroke (PO1-NS38475) (HCK, JJV), National Institute of Child Health and Development (Children’s Hospital Developmental Disabilities Research Center) (P30-HD18655), and the William Randolph Hearst Award.

References

  1. 1.
    Askalan R, Deveber G, Ho M, Ma J, Hawkins C (2006) Astrocytic-inducible nitric oxide synthase in the ischemic developing human brain. Pediatr Res 60:687–692. doi:10.1203/01.pdr.0000246226.89215.a6 PubMedCrossRefGoogle Scholar
  2. 2.
    Back SA, Luo NL, Mallinson RA et al (2005) Selective vulnerability of preterm white matter to oxidative damage defined by F2-isoprostanes. Ann Neurol 58:108–120. doi:10.1002/ana.20530 PubMedCrossRefGoogle Scholar
  3. 3.
    Billiards SS, Haynes RL, Folkerth RD et al (2008) Myelin abnormalities without oligodendrocyte loss in periventricular leukomalacia. Brain Pathol 18:153–163. doi:10.1111/j.1750-3639.2007.00107.x PubMedCrossRefGoogle Scholar
  4. 4.
    Brown GC (2007) Mechanisms of inflammatory neurodegeneration: iNOS and NADPH oxidase. Biochem Soc Trans 35:1119–1121. doi:10.1042/BST0351166 PubMedCrossRefGoogle Scholar
  5. 5.
    Contestabile A (2000) Roles of NMDA receptor activity and nitric oxide production in brain development. Brain Res Brain Res Rev 32:476–509. doi:10.1016/S0165-0173(00)00018-7 PubMedCrossRefGoogle Scholar
  6. 6.
    Counsell SJ, Allsop JM, Harrison MC et al (2003) Diffusion-weighted imaging of the brain in preterm infants with focal and diffuse white matter abnormality. Pediatrics 112:1–7. doi:10.1542/peds.112.1.1 PubMedCrossRefGoogle Scholar
  7. 7.
    Dammann O, Kuban KC, Leviton A (2002) Perinatal infection, fetal inflammatory response, white matter damage, and cognitive limitations in children born preterm. Ment Retard Dev Disabil Res Rev 8:46–50. doi:10.1002/mrdd.10005 PubMedCrossRefGoogle Scholar
  8. 8.
    De Vries LS, Connell JA, Dubowitz LM, Oozeer RC, Dubowitz V, Pennock JM (1987) Neurological, electrophysiological and MRI abnormalities in infants with extensive cystic leukomalacia. Neuropediatrics 18:61–66. doi:10.1055/s-2008-1052453 PubMedCrossRefGoogle Scholar
  9. 9.
    Endoh M, Maiese K, Wagner J (1994) Expression of the inducible form of nitric oxide synthase by reactive astrocytes after transient global ischemia. Brain Res 651:92–100. doi:10.1016/0006-8993(94)90683-1 PubMedCrossRefGoogle Scholar
  10. 10.
    Flodmark O, Lupton B, Li D et al (1989) MR imaging of periventricular leukomalacia in childhood. AJR Am J Roentgenol 152:583–590PubMedGoogle Scholar
  11. 11.
    Folkerth RD, Keefe RJ, Haynes RL, Trachtenberg FL, Volpe JJ, Kinney HC (2004) Interferon-gamma expression in periventricular leukomalacia in the human brain. Brain Pathol 14:265–274PubMedGoogle Scholar
  12. 12.
    Folkerth RD, Trachtenberg FL, Haynes RL (2008) Oxidative injury in the cerebral cortex and subplate neurons in periventricular leukomalacia. J Neuropathol Exp Neurol 67:677–686. doi:10.1097/NEN.0b013e31817e5c5e PubMedCrossRefGoogle Scholar
  13. 13.
    Goncalves LF, Chaiworapongsa T, Romero R (2002) Intrauterine infection and prematurity. MRDD Res Rev 8:3–13Google Scholar
  14. 14.
    Haynes RL, Folkerth RD, Keefe RJ et al (2003) Nitrosative and oxidative injury to premyelinating oligodendrocytes in periventricular leukomalacia. J Neuropathol Exp Neurol 62:441–450PubMedGoogle Scholar
  15. 15.
    Haynes RL, Borenstein NS, Desilva TM et al (2005) Axonal development in the cerebral white matter of the human fetus and infant. J Comp Neurol 484:156–167. doi:10.1002/cne.20453 PubMedCrossRefGoogle Scholar
  16. 16.
    Haynes RL, Billiards SS, Borenstein NS, Volpe JJ, Kinney HC (2008) Diffuse axonal injury in periventricular leukomalacia as determined by apoptotic marker fractin. Pediatr Res 63:656–661. doi:10.1203/PDR.0b013e31816c825c PubMedCrossRefGoogle Scholar
  17. 17.
    Hindley S, Juurlink BH, Gysbers JW, Middlemiss PJ, Herman MA, Rathbone MP (1997) Nitric oxide donors enhance neurotrophin-induced neurite outgrowth through a cGMP-dependent mechanism. J Neurosci Res 47:427–439. doi:10.1002/(SICI)1097-4547(19970215)47:4<427::AID-JNR8>3.0.CO;2-G PubMedCrossRefGoogle Scholar
  18. 18.
    Ikeno S, Nagata N, Yoshida S, Takahashi H, Kigawa J, Terakawa N (2000) Immature brain injury via peroxynitrite production induced by inducible nitric oxide synthase after hypoxia-ischemia in rats. J Obstet Gynaecol Res 26:227–234PubMedCrossRefGoogle Scholar
  19. 19.
    Inder TE, Wells SJ, Mogridge NB, Spencer C, Volpe JJ (2003) Defining the nature of the cerebral abnormalities in the premature infant: a qualitative magnetic resonance imaging study. J Pediatr 143:171–179. doi:10.1067/S0022-3476(03)00357-3 PubMedCrossRefGoogle Scholar
  20. 20.
    Kadhim H, Tabarki B, Verellen G, De Prez C, Rona A-M, Sebire G (2001) Inflammatory cytokines in the pathogenesis of periventricular leukomalacia. Neurology 56:1278–1284PubMedGoogle Scholar
  21. 21.
    Kadhim H, Tabarki B, De Prez C, Sebire G (2003) Cytokine immunoreactivity in cortical and subcortical neurons in periventricular leukomalacia: are cytokines implicated in neuronal dysfunction in cerebral palsy? Acta Neuropathol 105:209–216PubMedGoogle Scholar
  22. 22.
    Kadhim H, Khalifa M, Deltenre P, Casimir G, Sebire G (2006) Molecular mechanisms of cell death in periventricular leukomalacia. Neurology 67:293–299. doi:10.1212/01.wnl.0000224754.63593.c4 PubMedCrossRefGoogle Scholar
  23. 23.
    Katsuse O, Iseki E, Kosaka K (2003) Immunohistochemical study of the expression of cytokines and nitric oxide synthases in brains of patients with dementia with Lewy bodies. Neuropathology 23:9–15. doi:10.1046/j.1440-1789.2003.00483.x PubMedCrossRefGoogle Scholar
  24. 24.
    Kaur C, Sivakumar V, Ang LS, Sundaresan A (2006) Hypoxic damage to the periventricular white matter in neonatal brain: role of vascular endothelial growth factor, nitric oxide and excitotoxicity. J Neurochem 98:1200–1216. doi:10.1111/j.1471-4159.2006.03964.x PubMedCrossRefGoogle Scholar
  25. 25.
    Korhonen R, Linker K, Pautz A, Forstermann U, Moilanen E, Kleinert H (2007) Post-transcriptional regulation of human inducible nitric-oxide synthase expression by the Jun N-terminal kinase. Mol Pharmacol 71:1427–1434. doi:10.1124/mol.106.033449 PubMedCrossRefGoogle Scholar
  26. 26.
    Li J, Baud O, Vartanian T, Volpe JJ, Rosenberg PA (2005) Peroxynitrite generated by inducible nitric oxide synthase and NADPH oxidase mediates microglial toxicity to oligodendrocytes. Proc Natl Acad Sci USA 102:9936–9941. doi:10.1073/pnas.0502552102 PubMedCrossRefGoogle Scholar
  27. 27.
    Liu JS, Zhao ML, Brosnan CF, Lee SC (2001) Expression of inducible nitric oxide synthase and nitrotyrosine in multiple sclerosis lesions. Am J Pathol 158:2057–2066PubMedGoogle Scholar
  28. 28.
    Maalouf EF, Duggan PJ, Counsell S et al (2001) Comparison of findings on cranial ultrasound and magnetic resonance imaging in preterm infants. Pediatrics 107:719–727. doi:10.1542/peds.107.4.719 PubMedCrossRefGoogle Scholar
  29. 29.
    Muller CM (1992) Astrocytes in cat visual cortex studied by GFAP and S-100 immunocytochemistry during postnatal development. J Comp Neurol 317:309–323. doi:10.1002/cne.903170308 PubMedCrossRefGoogle Scholar
  30. 30.
    Nicholson S, Bonecini-Almeida Mda G, Lapa e Silva JR et al (1996) Inducible nitric oxide synthase in pulmonary alveolar macrophages from patients with tuberculosis. J Exp Med 183:2293–2302. doi:10.1084/jem.183.5.2293 PubMedCrossRefGoogle Scholar
  31. 31.
    Niwa M, Inao S, Takayasu M et al (2001) Time course of expression of three nitric oxide synthase isoforms after transient middle cerebral artery occlusion in rats. Neurol Med Chir (Tokyo) 41:63–73. doi:10.2176/nmc.41.63 CrossRefGoogle Scholar
  32. 32.
    Oleszak EL, Zaczynska E, Bhattacharjee M, Butunoi C, Legido A, Katsetos CD (1998) Inducible nitric oxide synthase and nitrotyrosine are found in monocytes/macrophages and/or astrocytes in acute, but not in chronic, multiple sclerosis. Clin Diagn Lab Immunol 5:438–445PubMedGoogle Scholar
  33. 33.
    Pautz A, Linker K, Hubrich T, Korhonen R, Altenhofer S, Kleinert H (2006) The polypyrimidine tract-binding protein (PTB) is involved in the post-transcriptional regulation of human inducible nitric oxide synthase expression. J Biol Chem 281:32294–32302. doi:10.1074/jbc.M603915200 PubMedCrossRefGoogle Scholar
  34. 34.
    Pierson CR, Folkerth RD, Billiards SS et al (2007) Gray matter injury associated with periventricular leukomalacia in the premature infant. Acta Neuropathol 114:619–631. doi:10.1007/s00401-007-0295-5 PubMedCrossRefGoogle Scholar
  35. 35.
    Renteria RC, Constantine-Paton M (1996) Exogenous nitric oxide causes collapse of retinal ganglion cell axonal growth cones in vitro. J Neurobiol 29:415–428. doi:10.1002/(SICI)1097-4695(199604)29:4<415::AID-NEU1>3.0.CO;2-B PubMedCrossRefGoogle Scholar
  36. 36.
    Rochefort N, Quenech’du N, Watroba L, Mallat M, Giaume C, Milleret C (2002) Microglia and astrocytes may participate in the shaping of visual callosal projections during postnatal development. J Physiol (Paris) 96:183–192. doi:10.1016/S0928-4257(02)00005-0 CrossRefGoogle Scholar
  37. 37.
    Rochefort N, Quenech’du N, Ezan P, Giaume C, Milleret C (2005) Postnatal development of GFAP, connexin43 and connexin30 in cat visual cortex. Brain Res Dev Brain Res 160:252–264. doi:10.1016/j.devbrainres.2005.09.011 PubMedCrossRefGoogle Scholar
  38. 38.
    Shah DK, Doyle LW, Anderson PJ et al (2008) Adverse neurodevelopment in preterm infants with postnatal sepsis or necrotizing enterocolitis is mediated by white matter abnormalities on magnetic resonance imaging at term. J Pediatr 153:170–175. doi:10.1016/j.jpeds.2008.02.033 PubMedCrossRefGoogle Scholar
  39. 39.
    Towfighi J, Zec N, Yager J, Housman C, Vannucci RC (1995) Temporal evolution of neuropathologic changes in an immature rat model of cerebral hypoxia: a light microscopic study. Acta Neuropathol 90:375–386. doi:10.1007/BF00315011 PubMedCrossRefGoogle Scholar
  40. 40.
    Van Wagenen S, Rehder V (2001) Regulation of neuronal growth cone filopodia by nitric oxide depends on soluble guanylyl cyclase. J Neurobiol 46:206–219. doi:10.1002/1097-4695(20010215)46:3<206::AID-NEU1003>3.0.CO;2-S PubMedCrossRefGoogle Scholar
  41. 41.
    Volpe JJ (2001) Neurobiology of periventricular leukomalacia in the premature infant. Pediatr Res 50:553–562. doi:10.1203/00006450-200111000-00003 PubMedCrossRefGoogle Scholar
  42. 42.
    Volpe JJ (2008) Postnatal sepsis, necrotizing entercolitis, and the critical role of systemic inflammation in white matter injury in premature infants. J Pediatr 153:160–163. doi:10.1016/j.jpeds.2008.04.057 PubMedCrossRefGoogle Scholar
  43. 43.
    Wang MX, Murrell DF, Szabo C, Warren RF, Sarris M, Murrell GA (2001) Nitric oxide in skeletal muscle: inhibition of nitric oxide synthase inhibits walking speed in rats. Nitric Oxide 5:219–232. doi:10.1006/niox.2001.0348 PubMedCrossRefGoogle Scholar
  44. 44.
    Wong A, Luth HJ, Deuther-Conrad W et al (2001) Advanced glycation endproducts co-localize with inducible nitric oxide synthase in Alzheimer’s disease. Brain Res 920:32–40. doi:10.1016/S0006-8993(01)02872-4 PubMedCrossRefGoogle Scholar
  45. 45.
    You Y, Kaur C (2000) Expression of induced nitric oxide synthase in amoeboid microglia in postnatal rats following an exposure to hypoxia. Neurosci Lett 279:101–104. doi:10.1016/S0304-3940(99)00967-2 PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Robin L. Haynes
    • 1
    • 3
  • Rebecca D. Folkerth
    • 1
    • 2
  • Felicia L. Trachtenberg
    • 4
  • Joseph J. Volpe
    • 3
    • 5
  • Hannah C. Kinney
    • 1
    • 3
  1. 1.Departments of PathologyChildren’s Hospital BostonBostonUSA
  2. 2.Brigham and Women’s HospitalBostonUSA
  3. 3.Harvard Medical SchoolBostonUSA
  4. 4.New England Research InstitutesWatertownUSA
  5. 5.Department of NeurologyChildren’s Hospital BostonBostonUSA

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