Acta Neuropathologica

, Volume 86, Issue 1, pp 1–9 | Cite as

Fluorescent histochemical localization of lipid peroxidation during brain reperfusion following cardiac arrest

  • Blaine C. White
  • Asif Daya
  • Donald J. DeGracia
  • Brian J. O'Neil
  • John M. Skjaerlund
  • Scott Trumble
  • Gary S. Krause
  • José A. Rafols
Regular Papers

Summary

Rats were subjected to cardiac arrest and resuscitation, 90 min of reperfusion, and in situ perfusion fixation. Thiobarbituric acid (TBA) was included in the aldehyde-free perfusion fixative, the TBA reaction was driven in situ by heating, and fluorescence microscopy was utilized to characterize the location of products of the TBA reaction. Absorbance-difference spectra were performed on butanol-extracted brain homogenates to confirm in situ formation of TBA adducts with aldehydic products of lipid peroxidation. Nissl-stained sections revealed good cellular fixation without shrinkage artifacts. Fluorescence was not seen microscopically when TBA was omitted from the perfusion fixative, and little fluorescence was present in normal brains or brains after ischemia only. However, after 90-min reperfusion, intense granular fluorescence was seen in the neuronal perikarya (especially at the base of the apical dendrite) of numerous pyramidal neurons in cortical layers 5 and 6 and in the pyramidal layer of Ammon's horn in the hippocampus. The nuclei of these cells exhibited no fluorescence. Fluorescence was also present in some striatal neurons, but was absent in the adjacent radial bundles. Neither glia nor white matter exhibited similar fluorescence. These observations indicate that neurons in the selectively vulnerable zones of the cortex and hippocampus are early and specific targets of lipid peroxidation during post-ischemic reperfusion.

Key words

Cerebral ischemia Lipid peroxidation Pathology Reperfusion 

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References

  1. 1.
    Akiguchi I, Horie R, Yamori Y (1980) Role of central aminergic fibers in experimental cerebral ischemia in strokeprone SHR. Stroke 11:383–389Google Scholar
  2. 2.
    Amaral DG, Imsauti R, Campbell MJ (1988) Distribution of somatostatin immunoreactivity in the human dentate gyrus. j Neurosci 8:3306–3316Google Scholar
  3. 3.
    Babbs CF, Ralston SH, Badylak SF et al (1986) Evidence of preventable lipid peroxidation in brain and heart four hours after cardiac arrest and reperfusion in rats (abstract). Am J Emerg Med 4:414Google Scholar
  4. 4.
    Bromont C, Marie C, Bralet J (1989) Increased lipid peroxidation in vulnerable brain regions after transient forebrain ischemia in rats. Stroke 20:918–924Google Scholar
  5. 5.
    Chan PH, Chen SF, Yu ACH (1988) Induction of intracellular superoxide radical formation by arachidonic acid and by polyunsaturated fatty acids in primary astrocytic cultures. J Neurochem 50:1185–1193Google Scholar
  6. 6.
    Chapat S, Frey V, Claperon N, Bouchaud C, Puisieux F, Couvreur P, Rossignol P, Delattre J (1991) Efficiency of liposomal ATP in cerebral ischemia: bioavailability features. Brain Res Bull 26:339–342Google Scholar
  7. 7.
    Contreras IR, Bender RA, Mansour J, Henry S, Shapiro L (1979) Caulobacter cresentus mutant defective in membrane phospholipid synthesis. J Bacteriol 140:612–619Google Scholar
  8. 8.
    Cooper RA, Ip SH, Cassileth PA, Kuo AL (1981) Inhibition of sterol and phospholipid synthesis in HL-60 promyelocytic leukemia cells by inducers of myeloid differentiation. Cancer Res 4:1847–1852Google Scholar
  9. 9.
    Csiba L, Paschen W, Hossmann KA (1983) A topographic quantitative method for measuring brain tissue pH under physiological and pathophysiological conditions. Brain Res 289:334–337Google Scholar
  10. 10.
    Czyrko C, Steigman C, Turley DL, Drott HR, Ziegler MM (1991) The role of reperfusion injury in occlusive intestinal ischemia of the neonate: Malonaldehyde-derived fluorescent products and correlation of histology. J Surg Res 51:1–4Google Scholar
  11. 11.
    de Lanerolle NC, Kim TH, Robbins RJ, Spencer DD (1989) Hippocampal interneuron loss and plasticity in human temporal lobe epilepsy. Brain Res 5:387–395Google Scholar
  12. 12.
    Demopoulos HB, Flamm ES, Pietronigro DD, Seligman ML (1980) The free radical pathology and the microcirculation in the major central nervous system disorders. Acta Physiol Scand 492 [Suppl]:91–95Google Scholar
  13. 13.
    Dianzani U (1989) Lipid peroxidation and cancer: a critical reconsideration. Tumori 75:351–357Google Scholar
  14. 14.
    Esterbauer H, Cheeseman KH (1990) Determination of aldehydic lipid peroxidation products: malondialdehyde and 4-hydroxynonenal. Methods Enzymol 186:407–421Google Scholar
  15. 15.
    Farquhar MG (1978) Traffic of products and membranes through the Golgi complex. In: Silverstein KSC (ed) Transport of macromolecules in cellular systems. Dalilem Konferenzen, Berlin, pp 341–362Google Scholar
  16. 16.
    Folbergrová J, Kiyota Y, Pahlmark K, Memezawa H, Smith ML, Siesjö BK (1993) Does ischemia with reperfusion lead to oxidative damage to proteins in the brain? J Cereb Blood Flow Metab 13:145–152Google Scholar
  17. 17.
    Gutteridge JMC, Torg D (1982) Iron-dependent free radical damage to DNA and deoxyribose: separation of TBA-reactive intermediates. Int J Biochem 14:891–893Google Scholar
  18. 18.
    Hoehner TJ, Garriatano AM, DiLorenzo RA, O'Neil BJ, Kumar K, Koehler J, Nayini N, Huang RR, Krause GS, Aust SD, White BC (1987) Brain cortex tissue Ca, Mg, Fe, Na and K following resuscitation from a 15-minute cardiac arrest in dogs. Am J Emerg Med 5:19–25Google Scholar
  19. 19.
    Janero DR (1990) Malondialdhyde and thiobarbituric acidreactivity as diagnostic indices of lipid peroxidation and peroxidative tissue injury. Free Radic Biol Med 9:515–540Google Scholar
  20. 20.
    Jenkins LW, Povlishock JT, Lewelt W, Miller JD, Becker DP (1981) The role of post-ischemic recirculation in the development of ischemic neuronal injury following complete cerebral ischemia. Acta Neuropathol (Berl) 55:205–220Google Scholar
  21. 21.
    Joseleau-Petit D, Kepes F, Peutat L, D'Ari R, Kepes A (1987) DNA replication initiation, doubling of rate of phospholipid synthesis, and cell division in E. coli. J Bacteriol 169:3701–3706Google Scholar
  22. 22.
    Kogure K, Watson BD, Busto R, Abe K (1982) Potentiation of lipid peroxides by ischemia in rat brain. Neurochem Res 7:437–454Google Scholar
  23. 23.
    Kohler C, Eriksson LG, Davies S, Chan-Palay V (1987) Co-localization of neuropeptide tyrosine and somatostatin immunoreactivity in neurons of individual subfields of the rat hippocampal region. Neurosci Lett 78:1–6Google Scholar
  24. 24.
    Kosugi H, Kukigawa K (1985) Thiobarbituric acid reaction of aldehydes and oxidized lipids in glacial acetic acid. Lipids 20:915–921Google Scholar
  25. 25.
    Kosugi H, Kojima T, Kikugawa K (1989) Thiobarbituric acid-reactive substances from peroxidized lipids. Lipids 24:873–881Google Scholar
  26. 26.
    Krause GS, DeGracia DJ, Skjaerlund JM, O'Neil BJ (1992) Assessment of free radical-induced damage in brain proteins after ischemia and reperfusion. Resuscitation 23:59–69Google Scholar
  27. 27.
    Kumar K, Goosman M, Krause GS, Nayini NR, Estrada R, Hoenher TJ, White BC, Koestner A (1987) Ultrastructural and ionic changes in global ischemic dog brain. Acta Neuropathol (Berl) 73:393–399Google Scholar
  28. 28.
    Marinari UM, Pronzato MA, Cottalasso D, Rolla C, Biasi F, Poli G, Nanni G, Dianzani MU (1987) Inhibition of liver Golgi glycosylation activities by carbonyl products of lipid peroxidation. Free Radic Res Commun 3:319–324Google Scholar
  29. 29.
    Nayini NR, White BC, Aust SD, Huang RR, Indrieri RJ, Evans AT, Bialek H, Jacobs WA, Komara J (1985) Post resuscitation iron delocalization and malondialdehyde production in the brain following prolonged cardiac arrest. J Free Radic Biol Med 1:111–116Google Scholar
  30. 30.
    Negovskii VA (1962) Resuscitation and artificial hypothermia. Consultants Bureau, New YorkGoogle Scholar
  31. 31.
    O'Neil BJ, Krause GS, White BC (1991) Thymine glycols and pyrmidine dimers in brain DNA during post-ischemic reperfusion. Resuscitation 21:41–55Google Scholar
  32. 32.
    Paschen W, Djuricic BM, Bosma HJ, Hossmann KA (1983) Biochemical changes during graded brain ischemia in gerbils. 2. Regional evaluation of cerebral blood flow and brain metabolites. J Neurol Sci 58:37–44Google Scholar
  33. 33.
    Paxinos G, Watson C (1986) The rat brain in stereotaxic coordinates, 2nd edn. Academic Press, New YorkGoogle Scholar
  34. 34.
    Peters A, Palay SL, deWebster H (1991) The fine structure of the nervous system: neurons and their supporting cells, 3rd edn. Oxford University Press, New York, pp 21–22Google Scholar
  35. 35.
    Petito CK, Pulsinelli WA (1984) Sequential development of reversible and irreversible neuronal damage following cerebral ischemia. J Neuropathol Exp Neurol 43:141–153Google Scholar
  36. 36.
    Pronzato MA, Cottalasso D, Dominicotti C, Tenca C, Traverso N, Nanni G, Marinari UM (1990) Effects of CCl4 poisoning on metabolism of dolichol in rat liver microsomes and Golgi apparatus. Free Radic Res Commun 11:267–277Google Scholar
  37. 37.
    Recknagel RO, Ghoshal AK (1966) Quantitative estimation of peroxidative degeneration of rat liver microsomal and mitochondrial lipids after carbon tetrachloride poisoning. Exp Mol Pathol 5:413–426Google Scholar
  38. 38.
    Rosenthal RE, Chanderbhan R, Marshall G, Fiskum G (1992) Prevention of post-ischemic brain lipid conjugated diene production and neurological injury by hydroxyethal starchconjugated deforoxamine. Free Radic Biol Med 12:29–33Google Scholar
  39. 39.
    Sakamoto A, Ohnishi ST, Ohnishi T, Ogawa R (1991) Relationship between free radical production and lipid peroxidation during ischemia-reperfusion injury in rat brain. Brain Res 554:186–192Google Scholar
  40. 40.
    Sato M, Hashimoto H, Kosaka F (1990) Histological changes of neuronal damage in vegetative dogs induced by 18 minutes of complete global brain ischemia: two-phase damage of Purkinge cells and hippocampal CA1 pyramidel cells. Acta Neuropathol 80:527–534Google Scholar
  41. 41.
    Shimasaki H, Hirai N, Ueta N (1988) Comparison of fluorescence characteristics of products of peroxidation of membrane phospholipids with those of products derived from reaction of malonaldehyde with glycine as a model of lipofuscin fluorescent substances. J Biochem 104:761–766Google Scholar
  42. 42.
    Siperstein MD (1984) Role of cholesterogenesis and isoprenoid synthesis in DNA replication and cell growth. J Lipid Res 25:1462–1468Google Scholar
  43. 43.
    Skjaerlund JM, Krause GS, Feldman DM, White BC (1991) The effect of EMHP on post-cardiac arrest survival of rats. Resuscitation 22:139–149Google Scholar
  44. 44.
    Umemura A (1990) Regional difference in free fatty acids release and the action of phospholipase during ischemia in rat brain. No To Shinkei 42:979–986Google Scholar
  45. 45.
    Voelker DR (1990) Lipid transport pathways in mammalian cells. Experientia 46:569–579Google Scholar
  46. 46.
    Watson BD, Busto R, Goldberg WJ, Santiso M, Yoshida S, Ginsberg MD (1984) Lipid peroxidation in vivo induced by reversible global ischemia in rat brain. J Neurochem 42:268–274Google Scholar
  47. 47.
    Weinberger J, Nieves-Rosa J (1988) Monoamine neurotransmitters in the evolution of infarction in ischemic striatum: morphologic correlation. J Neural Transm 71:133–142Google Scholar
  48. 48.
    Welsh FA, Marcy VR, Sims RE (1991) NADH fluorescence and regional energy metabolites during focal ischemia and reperfusion of rat brain. J Cereb Blood Flow Metab 11:459–465Google Scholar
  49. 49.
    Whaley WG, Dauwalder M (1979) The Golgi apparatus, the plasma membrane, and functional integration. Int Rev Cytol 58:199–245Google Scholar
  50. 50.
    White BC, Nayini NR, Krause GS, Aust SD, March GG, Bicknell JS IV, Goosmann M (1988) Effect on biochemical markers of brain injury of therapy with deferoxamine or superoxide dismutase following cardiac arrest. Am J Emerg Med 6:569–576Google Scholar
  51. 51.
    White BC, DeGracia DJ, Krause GS, Skjaerlund JM, O'Neil BJ, Grossman LI (1991) Brain nuclear DNA survives cardiac arrest and reperfusion. J Free Radic Biol Med 10:125–135Google Scholar
  52. 52.
    White BC, Tribhuwan RC, Vander Laan DJ, DeGracia DJ, Krause GS, Grossman LI (1992) Brain mitochondrial DNA is not damaged by prolonged cardiac arrest or reperfusion. J Neurochem 58:1716–1722Google Scholar
  53. 53.
    Yoshida S, Inoh S, Asano T, Sano K, Kubota M, Shimazaki H, Ueta N (1980) Effect of transient ischemia on free fatty acids and phospholipids in the gerbil brain. J Neurosurg 53:323–331Google Scholar
  54. 54.
    Zaleska MM, Floyd RA (1985) Regional lipid peroxidation in rat brain in vitro: possible role of endogenous iron. Neurochem Res 10:397–410Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Blaine C. White
    • 1
  • Asif Daya
    • 2
  • Donald J. DeGracia
    • 1
  • Brian J. O'Neil
    • 1
  • John M. Skjaerlund
    • 1
  • Scott Trumble
    • 1
  • Gary S. Krause
    • 1
  • José A. Rafols
    • 2
  1. 1.Department of Emergency MedicineWayne State University School of MedicineDetroitUSA
  2. 2.Department of Anatomy and Cell BiologyWayne State University School of MedicineDetroitUSA

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