Acta Neuropathologica

, Volume 113, Issue 3, pp 277–293 | Cite as

Accelerated infarct development, cytogenesis and apoptosis following transient cerebral ischemia in aged rats

  • Aurel Popa-Wagner
  • Irina Badan
  • Lary Walker
  • Sergiu Groppa
  • Nicoleta Patrana
  • Christof Kessler
Original Paper


Old age is associated with a deficient recovery from stroke, but the cellular mechanisms underlying such phenomena are poorly understood. To address this issue, focal cerebral ischemia was produced by reversible occlusion of the right middle cerebral artery in 3- and 20-month-old male Sprague–Dawley rats. Aged rats showed a delayed and suboptimal functional recovery in the post-stroke period. Using BrdU-labeling, quantitative immunohistochemistry and 3-D reconstruction of confocal images, we found that aged rats are predisposed to rapidly develop an infarct within the first few days after ischemia. The emergence of the necrotic zone is associated with a high rate of cellular degeneration, premature accumulation of proliferating BrdU-positive cells that appear to emanate from capillaries in the infarcted area, and a large number of apoptotic cells. With double labeling techniques, we were able to identify, for the first time, over 60% of BrdU-positive cells either as reactive microglia (45%), oligodendrocyte progenitors (17%), astrocytes (23%), CD8+ lymphocytes (4%), or apoptotic cells (<1%). Paradoxically, despite a robust reactive phenotype of microglia and astrocytes in aged rats, at 1-week post-stroke, the number of proliferating microglia and astrocytes was lower in aged rats than in young rats. Our data indicate that aging is associated with rapid infarct development and a poor prognosis for full recovery from stroke that is correlated with premature cellular proliferation and increased cellular degeneration and apoptosis in the infarcted area.


Stroke Aging Recovery Ischemia Rat BrdU GFAP Oligodendrocytes Microglia CD8+ lymphocytes Cytogenesis Vascular tree 



This research was supported by a grant from Deutsche Forschungsgemeinschaft (DFG) to CK (Ke 599/1–1), by NIH RR-00165 (LCW) and by a grant from “Prof. Dieter Platt Stiftung” to APW.


  1. 1.
    Abdel-Rahman A, Rao MS, Shetty AK (2004) Nestin expression in hippocampal astrocytes after injury depends on the age of the hippocampus. Glia 47:299–313PubMedCrossRefGoogle Scholar
  2. 2.
    Asher RA, Morgenstern DA, Fidler PS, Adcock KH, Oohira A, Braistead JE, Levine JM, Margolis RU, Rogers JH, Fawcett JW (2002) Neurocan is upregulated in injured brain and in cytokine-treated astrocytes. J Neurosci 20:2427–2438Google Scholar
  3. 3.
    Adams MM, Shah RA, Janssen WG, Morrison JH (2001) Different modes of hippocampal plasticity in response to estrogen in young and aged female rats. Proc Natl Acad Sci USA 98:8071–8076PubMedCrossRefGoogle Scholar
  4. 4.
    Adams MM, Gazzaley AH, Morrison JH (2001) Attenuated lesion-induced N-methyl-d-aspartate receptor (NMDAR) plasticity in the dentate gyrus of aged rats following perforant path lesions. Exp Neurol 172:244–249PubMedCrossRefGoogle Scholar
  5. 5.
    Aliev G, Smith MA, Seyidov D, Neal ML, Lamb BT, Nunomura A, Gasimov EK, Vinters HV, Perry G, LaManna JC, Friedland RP (2002) The role of oxidative stress in the pathophysiology of cerebrovascular lesions in Alzheimer’s disease. Brain Pathol 12:21–35PubMedCrossRefGoogle Scholar
  6. 6.
    Badan I, Buchhold B, Hamm A, Gratz M, Walker LC, Platt D, Kessler Ch, Popa-Wagner A (2003) Accelerated glial reactivity to stroke in aged rats correlates with reduced functional recovery. J Cereb Blood Flow Metab 23:845–854PubMedCrossRefGoogle Scholar
  7. 7.
    Badan I, Dinca I, Buchhold B, Suofu Y, Walker L, Gratz M, Platt D, Kessler Ch, Popa-Wagner A (2004) Accelerated accumulation of N- and C-terminal betaAPP fragments and delayed recovery of microtubule-associated protein 1B expression following stroke in aged rats. Eur J Neurosci 19:2270–2280PubMedCrossRefGoogle Scholar
  8. 8.
    Barnett HJ (2002) Stroke prevention in the elderly. Clin Exp Hypertens 24:563–571PubMedCrossRefGoogle Scholar
  9. 9.
    Bury SD, Jones TA (2002) Unilateral sensorimotor cortex lesions in adult rats facilitate motor skill learning with the “unaffected” forelimb and training-induced dendritic structural plasticity in the motor cortex. J Neurosci 22:8597–8606PubMedGoogle Scholar
  10. 10.
    Bondolfi L, Calhoun M, Ermini F, Kuhn HG, Widerhold K-H, Walker L, Staufenbiel M, Jucker (2002) Amyloid-associated neuron loss and gliogenesis in the neocortex of amyloid precursor protein transgenic mice. J Neurosci 22:515–522PubMedGoogle Scholar
  11. 11.
    Brown AW, Marlowe KJ, Bjelke B (2003) Age effect on motor recovery in a post-acute animal stroke model. Neurobiol Aging 24:607–614PubMedCrossRefGoogle Scholar
  12. 12.
    Cameron HA, McKay RD (2001) Adult neurogenesis produces a large pool of new granule cells in the dentate gyrus. J Comp Neurol 435:406–417PubMedCrossRefGoogle Scholar
  13. 13.
    Chen ZJ, Ughrin Y, Levine JM (2002) Inhibition of axon growth by oligodendrocyte precursor cells. Mol Cell Neurosci 20:125–139PubMedCrossRefGoogle Scholar
  14. 14.
    Davies M, Mendelow AD, Perry RH, Chambers IR, James OFW (1995) Experimental stroke and neuroprotection in the aging rat brain. Stroke 26:1072–1078Google Scholar
  15. 15.
    Dewar D, Underhill SM, Goldberg MP (2003) Oligodendrocyte progenitors and ischemic brain injury. J Cereb Blood Flow Metab 23:263–274PubMedCrossRefGoogle Scholar
  16. 16.
    Floyd RA, Hensley K (2000) Nitrone inhibition of age-associated oxidative damage. Ann N Y Acad Sci 899:222–237PubMedCrossRefGoogle Scholar
  17. 17.
    Floyd RA, Hensley K (2002) Oxidative stress in brain aging. Implications for therapeutics of neurodegenerative diseases. Neurobiol Aging 23:795–807PubMedCrossRefGoogle Scholar
  18. 18.
    Frost SB, Barbay S, Friel KM, Plautz EJ, Nudo RJ (2003) Reorganization of remote cortical regions after ischemic brain injury: a potential substrate for stroke recovery. J Neurophysiol 89:3205–3214PubMedCrossRefGoogle Scholar
  19. 19.
    Garcia JH, Wagner S, Liu K-F, Hu X (1995) Neurological deficit and extent of neuronal necrosis attributable to middle cerebral artery occlusion in rats. Stroke 26:627–635PubMedGoogle Scholar
  20. 20.
    Gozal D, Row BW, Kheirandish L, Li R, Guo RL, Qiang F, Brittian KR (2003) Increased susceptibility to intermittent hypoxia in aging rats: changes in proteasomal activity, neuronal apoptosis and spatial function. J Neurochem 86:1545–1552PubMedCrossRefGoogle Scholar
  21. 21.
    Hajdu MA, Heistad DD, Siems JE, Baumbach GL (1990) Effects of aging on mechanics and composition of cerebral arterioles in rats. Circ Res 66:1747–1754PubMedGoogle Scholar
  22. 22.
    Hess DC, Hill WD, Carroll JE, Borlongan CV (2004) Do bone marrow cells generate neurons?. Arch Neurol 61:483–485PubMedCrossRefGoogle Scholar
  23. 23.
    Hiona A, Leeuwenburgh C (2004) Effects of age and caloric restriction on brain neuronal cell death/survival. Ann N Y Acad Sci 1019:96–105PubMedCrossRefGoogle Scholar
  24. 24.
    Hoane MR, Lasley LA, Akstulewicz SL (2004) Middle age increases tissue vulnerability and impairs sensorimotor and cognitive recovery following traumatic brain injury in the rat. Behav Brain Res 153:189–197PubMedCrossRefGoogle Scholar
  25. 25.
    Hoehn BD, Palmer TD, Steinberg GK (2005) Neurogenesis in rats after focal cerebral ischemia is enhanced by indomethacin. Stroke 36:2718–2724PubMedCrossRefGoogle Scholar
  26. 26.
    Hoff SF, Scheff SW, Cotman CW (1982) Lesion-induced synaptogenesis in the dentate gyrus of aged rats: I. Loss and reacquisition of normal synaptic density. J Comp Neurol 205:246–252PubMedCrossRefGoogle Scholar
  27. 27.
    Howard CV, Reed MG (1998) Unbiased stereology. BIOS Scientific Publishers Ltd, OxfordGoogle Scholar
  28. 28.
    Hsu JE, Jones TA (2006) Contralesional neural plasticity and functional changes in the less-affected forelimb after large and small cortical infarcts in rats. Exp Neurol. 2006 Jun 21; [Epub ahead of print]Google Scholar
  29. 29.
    Kempermann G, Gast D, Kronenberg G, Yamaguchi M, Gage FH (2003) Early determination and long-term persistence of adult-generated new neurons in the hippocampus of mice. Development 130:391–399PubMedCrossRefGoogle Scholar
  30. 30.
    Kuan CY, Schloemer AJ, Lu A, Burns KA, Weng WL, Williams MT, Strauss KI, Vorhees CV, Flavell RA, Davis RJ, Sharp FR, Rakic P (2004) Hypoxia-ischemia induces DNA synthesis without cell proliferation in dying neurons in adult rodent brain. J Neurosci 24:10763–10772PubMedCrossRefGoogle Scholar
  31. 31.
    Jones TA, Chu CJ, Grande LA, Gregory AD (1999) Motor skills training enhances lesion-induced structural plasticity in the motor cortex of adult rats. J Neurosci 19:10153–10163PubMedGoogle Scholar
  32. 32.
    Justicia C, Martin A, Rojas S, Gironella M, Cervera A, Panes J, Chamorro A, Planas AM (2005) Anti-VCAM-1 antibodies did not protect against ischemic damage either in rats or in mice. J Cereb Blood Flow Metab 26:421–432CrossRefGoogle Scholar
  33. 33.
    Li Y, Chen J, Chopp M (2001) Adult bone marrow transplantation after stroke in adultrats. Cell Transplant 10:31–40PubMedGoogle Scholar
  34. 34.
    Li Y, Chen J, Chopp M (2002) Cell proliferation and differentiation from ependymal, subependymal and choroid plexus cells in response to stroke in rats. J Neurol Sci 193:137–146PubMedCrossRefGoogle Scholar
  35. 35.
    Lindner MD, Gribkoff VK, Donlan NA, Jones TA (2003) Long-lasting functional disabilities in middle-aged rats with small cerebral infarcts. J Neurosci 23:10913–10922PubMedGoogle Scholar
  36. 36.
    Mabuchi T, Kitagawa K, Ohtsuki T, Kuwabara K, Yagita Y, Yanagihara T, Hori M, Maatsumoto M (2000) Contribution of microglia/macrophages to expansion of infarction and response of oligodendrocyte progenitors after focal cerebral ischemia in rats. Stroke 31:1735–1743PubMedGoogle Scholar
  37. 37.
    Markus TM, Tsai SY, Bollnow MR, Farrer RG, O’Brien TE, Kindler-Baumann DR,Rausch M, Rudin M, Wiessner C, Mir AK, Schwab ME, Kartje GL (2005) Recovery and brain reorganization after stroke in adult and aged rats. Ann Neurol 58:950–953PubMedCrossRefGoogle Scholar
  38. 38.
    Morgenstern DA, Asher RA, Fawcett JW (2002) Chondroitin sulphate proteoglycans in the CNS injury response. Prog Brain Res 137:313–332PubMedCrossRefGoogle Scholar
  39. 39.
    Nichols NR, Finch CE, Nelson JF (1995) Food restriction delays the age-related increase in GFAP mRNA in rat hypothalamus. Neurobiol Aging 16:105–110PubMedCrossRefGoogle Scholar
  40. 40.
    Ohta K, Iwai M, Sato K, Omori N, Nagano I, Shoji M, Abe K (2003) Dissociative increase of oligodendrocyte progenitor cells between young and aged rats after transient cerebral ischemia. Neurosci Lett 335:159–162PubMedCrossRefGoogle Scholar
  41. 41.
    Packard Jr DS, Menzies RA, Skalko RG (1973) Incorporation of thymidine and its analogue, bromodeoxyuridine, into embryos and maternal tissues of the mouse. Differentiation 1:397–404PubMedCrossRefGoogle Scholar
  42. 42.
    Parhad IM, Scott JN, Cellars LA, Bains JS, Krekoski CA, Clark AW (1995) Axonal atrophy in aging is associated with a decline in neurofilament gene expression. J Neurosci Res 41:355–366PubMedCrossRefGoogle Scholar
  43. 43.
    Petullo D, Masonic K, Lincoln C, Wibberley L, Teliska M, Yao DL (1999) Model development and behavioral assessment of focal cerebral ischemia in rats. Life Sci 64:1099–1108PubMedCrossRefGoogle Scholar
  44. 44.
    Popa-Wagner A, Schroder E, Walker LC, Kessler Ch (1998) Beta-Amyloid precursor protein and ss-amyloid peptide immunoreactivity in the rat brain after middle cerebral artery occlusion: effect of age. Stroke 29:2196–2202PubMedGoogle Scholar
  45. 45.
    Popa-Wagner A, Schröder E, Schmoll H, Walker LC, Kessler Ch (1999) Upregulation of MAP1B and MAP2 in the rat brain following middle cerebral artery occlusion: effect of age. J Cereb Blood Flow Metab 19:425–434PubMedCrossRefGoogle Scholar
  46. 46.
    Popa-Wagner A, Fischer B, Platt D, Neubig R, Schmoll H, Kessler C (1999) Anomalous expression of microtubule-associated protein 1B in the hippocampus and cortex of aged rats treated with pentylenetetrazole. Neuroscience 94: 395–403PubMedCrossRefGoogle Scholar
  47. 47.
    Popa-Wagner A, Fischer B, Platt D, Schmoll H, Kessler C (2000) Delayed and blunted induction of mRNA for tissue plasminogen activator in the brain of old rats following pentylenetetrazole-induced seizure activity. J Gerontol A Biol Sci Med Sci 55:B242–B248PubMedGoogle Scholar
  48. 48.
    Priller J, Persons DA, Klett FF, Kempermann G, Kreutzberg GW, Dirnagl U (2001) Neogenesis of cerebellar Purkinje neurons from gene-marked bone marrow cells in vivo. J Cell Biol 155:733–738PubMedCrossRefGoogle Scholar
  49. 49.
    Rakic P (2002) Adult neurogenesis in mammals: an identity crisis. J Neurosci 22:614–618PubMedGoogle Scholar
  50. 50.
    Retchkiman I, Fischer B, Platt D, Popa-Wagner A (1996) Seizure induced c-fos mRNA in the rat brain: comparison between young and aging animals. Neurobiol Aging 17:41–44PubMedCrossRefGoogle Scholar
  51. 51.
    Riddle DR, Sonntag WE, Lichtenwalner RJ (2003) Microvascular plasticity in aging. Ageing Res Rev 2:149–168PubMedCrossRefGoogle Scholar
  52. 52.
    Rivlin AS, Tator CH (1977) Objective clinical assessment of motor function after experimental spinal cord injury in the rat. J Neurosurg 47:577–581PubMedCrossRefGoogle Scholar
  53. 53.
    Roberts EL Jr, Chih CP, Rosenthal M (1997) Age-related changes in brain metabolism and vulnerability to anoxia. Adv Exp Med Biol 411:83–89PubMedGoogle Scholar
  54. 54.
    Schauwecker PE, Cheng HW, Serquinia RM, Mori N, McNeill TH (1995) Lesion-induced sprouting of commissural/associational axons and induction of GAP-43 mRNA in hilar and CA3 pyramidal neurons in the hippocampus are diminished in aged rats. J Neurosci 15:2462–2470PubMedGoogle Scholar
  55. 55.
    Schmued LC, Hopkins KJ (2000) Fluoro-Jade B: a high affinity fluorescent marker for the localization of neuronal degeneration. Brain Res 874:123–130PubMedCrossRefGoogle Scholar
  56. 56.
    Shetty AK, Rao MS, Hattiangady B, Zaman V, Shetty GA (2004) Hippocampal neurotrophin levels after injury: Relationship to the age of the hippocampus at the time of injury. J Neurosci Res 78:520–532PubMedCrossRefGoogle Scholar
  57. 57.
    Shimamura M, Garcia JM, Prough DS, Hellmich HL (2004) Laser capture microdissection and analysis of amplified antisense RNA from distinct cell populations of the young and aged rat brain: effect of traumatic brain injury on hippocampal gene expression. Brain Res Mol Brain Res 122:47–61PubMedCrossRefGoogle Scholar
  58. 58.
    Stoll G, Jander S, Schroeter M (1998) Inflammation and glial responses in ischemic brain lesions. Prog Neurobiol 56:149–171PubMedCrossRefGoogle Scholar
  59. 59.
    Stone DJ, Rozovsky I, Morgan TE, Anderson CP, Lopez LM, Shick J, Finch CE (2000) Effects of age on gene expression during estrogen-induced synaptic sprouting in the female rat. Exp Neurol 165:46–57PubMedCrossRefGoogle Scholar
  60. 60.
    Sutherland GR, Dix GA, Auer RN (1996) Effect of age in rodent models of focal and forebrain ischemia. Stroke 27:1663–1667PubMedGoogle Scholar
  61. 61.
    Tomimoto H, Ihara M, Wakita H, Ohtani R, Lin JX, Akiguchi I, Kinoshita M, Shibasaki H (2003) Chronic cerebral hypoperfusion induces white matter lesions and loss of oligodendroglia with DNA fragmentation in the rat. Acta Neuropathol (Berl) 106:527–534CrossRefGoogle Scholar
  62. 62.
    Vallieres L, Sawchenko PE (2003) Bone marrow-derived cells that populate the adult mouse brain preserve their hematopoietic identity. J Neurosci 23:5197–5207PubMedGoogle Scholar
  63. 63.
    Vogelgesang S, Schroeder E, Walker LC, Pahnke J, Naubereit A, Walther R, Stausske D, Warzok RW (2002) Activated microglia do not mediate the early deposition of Abeta in carriers of the apolipoprotein Eepsilon4 allele. Clin Neuropathol 21:99–106PubMedGoogle Scholar
  64. 64.
    Wang LC, Futrell N, Wang DZ, Chen FJ, Zhai QH, Schulz LR (1995) A reproducible model of middle cerebral infarcts, compatible with long-term survival, in aged rats. Stroke 26:2087–2090PubMedGoogle Scholar
  65. 65.
    Wilhelmsson U, Li L, Pekna M, Berthold CH, Blom S, Eliasson C, Renner O, Bushong E, Ellisman M, Morgan TE, Pekny M (2004) Absence of glial fibrillary acidic protein and vimentin prevents hypertrophy of astrocytic processes and improves post-traumatic regeneration. J Neurosci 24:5016–5021PubMedCrossRefGoogle Scholar
  66. 66.
    Woods AG, Guthrie KM, Kurlawalla MA, Gall CM (1998) Deafferentation-induced increases in hippocampal insulin-like growth factor-1 messenger RNA expression are severely attenuated in middle aged and aged rats. Neuroscience 83:663–668PubMedCrossRefGoogle Scholar
  67. 67.
    Yu WH, Go L, Guinn BA, Fraser PE, Westaway D, McLaurin J (2002) Phenotypic and functional changes in glial cells as a function of age. Neurobiol Aging 23:105–115PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Aurel Popa-Wagner
    • 1
  • Irina Badan
    • 1
  • Lary Walker
    • 2
  • Sergiu Groppa
    • 4
  • Nicoleta Patrana
    • 3
  • Christof Kessler
    • 1
  1. 1.Department of NeurologyUniversity of GreifswaldGreifswaldGermany
  2. 2.Yerkes National Primate Research Center and Department of NeurologyEmory UniversityAtlantaUSA
  3. 3.University of Medicine and PharmacyCraiovaRomania
  4. 4.Klinik für Neuropädiatrie, Universitätsklinikum Schleswig-HolsteinKielGermany

Personalised recommendations