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Molecular Biology Reports

, Volume 46, Issue 6, pp 6215–6230 | Cite as

Temporal changes in physiological and molecular markers in various brain regions following transient global ischemia in rats

  • Monika Kapoor
  • Sheetal Sharma
  • Rajat Sandhir
  • Bimla NehruEmail author
Original Article
  • 97 Downloads

Abstract

Several mechanisms are involved in the loss of cellular integrity and tissue destructions in various brain regions during ischemic insult. The affected brain employs various self-repair mechanisms during the poststroke recovery. Therefore, the current study involves time course changes in different brain regions following ischemia in terms of inflammation, oxidative stress and apoptosis for which a bilateral common carotid arteries occlusion model was chosen. The development of oxidative stress was seen with a marked increase in ROS and NO levels with concomitant decrease in GSH levels and also the activities of anti-oxidant enzymes. These alterations were accompanied with decreased levels of neurotransmitters and motor and cognitive deficits at various time points. Increased expressions of various pro-inflammatory cytokines and a decline in BDNF levels in hippocampal regions on 7th day post ischemia, suggesting their role in its pathogenesis. The restoration of BDNF and neurotransmitter levels along with significant decline in inflammatory cytokine levels 14th day onwards following ischemia in hippocampus suggested poststroke recovery. The extent of neuronal damage was found to be increased significantly on 7th day post ischemia as indicated by TUNEL assay and hematoxylin and eosin staining depicting enhanced number of pyknotic neurons in cortical and hippocampal regions. Cortical regions of the ischemic brains were severely affected while hippocampal regions showed significant poststroke recovery, which might attributed to the normalization of BDNF and pro-inflammatory cytokine levels. In conclusion, the present study established the central role of BDNF and pro-inflammatory cytokines in the poststroke recovery. Also, the cortical and hippocampal regions were found to be more susceptible for ischemic injury. As our results indicated, full recovery after ischemic injury in different brain regions was not achieved, therefore further studies with long-term recovery time are required to be conducted.

Graphic abstract

Keywords

Inflammation Transient cerebral ischemia Oxidative stress Memory dysfunction Neurotransmitters 

Abbreviations

TGCI

Transient global cerebral ischemia

ROS/RNS

Reactive oxygen species/reactive nitrogen species

DA

Dopamine

NE

Norepinephrine

5-HT

Sertonin

DTNB

5,5′-Dithiobis-(2-nitrobenzoic acid)

TTC

Triphenyltetrazolium chloride

ECD

Electrochemical detector

TL

Transfer latency

BDNF

Brain derived neurotrophic factor

LSD

Least significant difference

Notes

Acknowledgements

The study was carried out with the funds provided by Department of Science & Technology/Innovation in Science Pursuit for Inspired Research (DST/INSPIRE), India with IF no. IF130058.

Compliance with ethical standards

Conflict of interest

The authors do not have any conflict of interest in the manuscript.

Ethical approval

The authors have read and abided by the statement of the ethical standards for manuscripts submitted to this journal.

References

  1. 1.
    Simats A, García-Berrocoso T, Montaner J (2016) Neuroinflammatory biomarkers: from stroke diagnosis and prognosis to therapy. Biochim Biophys Acta 1862:411–424.  https://doi.org/10.1016/j.bbadis.2015.10.025 CrossRefGoogle Scholar
  2. 2.
    Woodruff TM, Thundyil J, Tang S-C et al (2011) Pathophysiology, treatment, and animal and cellular models of human ischemic stroke. Mol Neurodegener 6:11.  https://doi.org/10.1186/1750-1326-6-11 CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Lee J-M, Grabb MC, Zipfel GJ, Choi DW (2000) Brain tissue responses to ischemia. J Clin Invest 106:723–731.  https://doi.org/10.1172/JCI11003 CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Lee J-C, Won M-H (2014) Neuroprotection of antioxidant enzymes against transient global cerebral ischemia in gerbils. Anat Cell Biol 47:149–156.  https://doi.org/10.5115/acb.2014.47.3.149 CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Kalogeris T, Baines CP, Krenz M, Korthuis RJ (2012) Cell biology of ischemia/reperfusion injury. Int Rev Cell Mol Biol 298:229–317.  https://doi.org/10.1016/B978-0-12-394309-5.00006-7 CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Kalogeris T, Bao Y, Korthuis RJ (2014) Mitochondrial reactive oxygen species: a double edged sword in ischemia/reperfusion vs preconditioning. Redox Biol 2:702–714.  https://doi.org/10.1016/j.redox.2014.05.006 CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Esenwa CC, Elkind MS (2016) Inflammatory risk factors, biomarkers and associated therapy in ischaemic stroke. Nat Rev Neurol 12:594–604.  https://doi.org/10.1038/nrneurol.2016.125 CrossRefGoogle Scholar
  8. 8.
    Kelly PJ, Murphy S, Coveney S et al (2018) Anti-inflammatory approaches to ischaemic stroke prevention. J Neurol Neurosurg Psychiatry 89:211–218.  https://doi.org/10.1136/jnnp-2016-314817 CrossRefGoogle Scholar
  9. 9.
    Amantea D, Bagetta G (2017) Excitatory and inhibitory amino acid neurotransmitters in stroke: from neurotoxicity to ischemic tolerance. Curr Opin Pharmacol 35:111–119.  https://doi.org/10.1016/j.coph.2017.07.014 CrossRefGoogle Scholar
  10. 10.
    Gu W, Gu C, Jiang W, Wester P (2010) Neurotransmitter synthesis in poststroke cortical neurogenesis in adult rats. Stem Cell Res 4:148–154.  https://doi.org/10.1016/j.scr.2009.12.001 CrossRefGoogle Scholar
  11. 11.
    Berg C, Backström T, Winberg S et al (2013) Developmental exposure to fluoxetine modulates the serotonin system in hypothalamus. PLoS ONE 8:e55053.  https://doi.org/10.1371/journal.pone.0055053 CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Alcantara CC, García-Salazar LF, Silva-Couto MA et al (2018) Post-stroke BDNF concentration changes following physical exercise: a systematic review. Front Neurol 9:637.  https://doi.org/10.3389/fneur.2018.00637 CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Ji X-W, Wu C-L, Wang X-C et al (2014) Monoamine neurotransmitters and fibroblast growth factor-2 in the brains of rats with post-stroke depression. Exp Ther Med 8:159–164.  https://doi.org/10.3892/etm.2014.1674 CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Mergenthaler P, Dirnagl U, Meisel A (2004) Pathophysiology of stroke: lessons from animal models. Metab Brain Dis 19:151–167CrossRefGoogle Scholar
  15. 15.
    Bartsch T, Döhring J, Reuter S et al (2015) Selective neuronal vulnerability of human hippocampal CA1 neurons: lesion evolution, temporal course, and pattern of hippocampal damage in diffusion-weighted MR imaging. J Cereb Blood Flow Metab 35:1836–1845.  https://doi.org/10.1038/jcbfm.2015.137 CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Wang X, Michaelis EK (2010) Selective neuronal vulnerability to oxidative stress in the brain. Front Aging Neurosci 2:12.  https://doi.org/10.3389/fnagi.2010.00012 CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Cavaglia M, Dombrowski SM, Drazba J et al (2001) Regional variation in brain capillary density and vascular response to ischemia. Brain Res 910:81–93CrossRefGoogle Scholar
  18. 18.
    Wang X, Pal R, Chen X-W et al (2005) High intrinsic oxidative stress may underlie selective vulnerability of the hippocampal CA1 region. Brain Res Mol Brain Res.  https://doi.org/10.1016/j.molbrainres.2005.07.018 CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Sieber FE, Palmon SC, Traystman RJ, Martin LJ (1995) Global incomplete cerebral ischemia produces predominantly cortical neuronal injury. Stroke 26:2091–2095 (discussion 2096)CrossRefGoogle Scholar
  20. 20.
    Hara A, Yoshimi N, Hirose Y et al (1995) DNA fragmentation in granular cells of human cerebellum following global ischemia. Brain Res 697:247–250.  https://doi.org/10.1016/0006-8993(95)00902-3 CrossRefGoogle Scholar
  21. 21.
    Wang K, Damjanov I, Wan Y-JY (2010) The protective role of pregnane X receptor in lipopolysaccharide/D-galactosamine-induced acute liver injury. Lab Investig 90:257–265.  https://doi.org/10.1038/labinvest.2009.129 CrossRefGoogle Scholar
  22. 22.
    Ouyang Y-B, Voloboueva LA, Xu L-J, Giffard RG (2007) Selective dysfunction of hippocampal CA1 astrocytes contributes to delayed neuronal damage after transient forebrain ischemia. J Neurosci 27:4253–4260.  https://doi.org/10.1523/JNEUROSCI.0211-07.2007 CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Kirino T, Sano K (1984) Selective vulnerability in the gerbil hippocampus following transient ischemia. Acta Neuropathol 62:201–208.  https://doi.org/10.1007/BF00691853 CrossRefGoogle Scholar
  24. 24.
    Jingtao J, Sato S, Yamanaka N (1999) Changes in cerebral blood flow and blood brain barrier in the gerbil hippocampal CA1 region following repeated brief cerebral ischemia. Med Electron Microsc 32:175–183.  https://doi.org/10.1007/s007959900012 CrossRefGoogle Scholar
  25. 25.
    Himori N, Watanabe H, Akaike N et al (1990) Cerebral ischemia model with conscious mice. Involvement of NMDA receptor activation and derangement of learning and memory ability. J Pharmacol Methods 23:311–327CrossRefGoogle Scholar
  26. 26.
    Morris RG, Garrud P, Rawlins JN, O’Keefe J (1982) Place navigation impaired in rats with hippocampal lesions. Nature 297:681–683CrossRefGoogle Scholar
  27. 27.
    Itoh J, Nabeshima T, Kameyama T (1991) Utility of an elevated plus-maze for dissociation of amnesic and behavioral effects of drugs in mice. Eur J Pharmacol 194:71–76CrossRefGoogle Scholar
  28. 28.
    Gaur V, Aggarwal A, Kumar A (2009) Protective effect of naringin against ischemic reperfusion cerebral injury: possible neurobehavioral, biochemical and cellular alterations in rat brain. Eur J Pharmacol 616:147–154.  https://doi.org/10.1016/j.ejphar.2009.06.056 CrossRefGoogle Scholar
  29. 29.
    Bishnoi M, Chopra K, Kulkarni SK (2006) Involvement of adenosinergic receptor system in an animal model of tardive dyskinesia and associated behavioural, biochemical and neurochemical changes. Eur J Pharmacol 552:55–66.  https://doi.org/10.1016/j.ejphar.2006.09.010 CrossRefGoogle Scholar
  30. 30.
    Church WH (2005) Column chromatography analysis of brain tissue: an advanced laboratory exercise for neuroscience majors. J Undergrad Neurosci Educ 3:A36–A41PubMedPubMedCentralGoogle Scholar
  31. 31.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275PubMedPubMedCentralGoogle Scholar
  32. 32.
    Best TM, Fiebig R, Corr DT et al (1999) Free radical activity, antioxidant enzyme, and glutathione changes with muscle stretch injury in rabbits. J Appl Physiol 87:74–82CrossRefGoogle Scholar
  33. 33.
    Raddassi K, Berthon B, Petit J-F, Lemaire G (1994) Role of calcium in the activation of mouse peritoneal macrophages: induction of NO synthase by calcium ionophores and thapsigargin. Cell Immunol 153:443–455.  https://doi.org/10.1006/cimm.1994.1041 CrossRefGoogle Scholar
  34. 34.
    Ellman GL (1959) Tissue sulfhydryl groups. Arch Biochem Biophys 82:70–77CrossRefGoogle Scholar
  35. 35.
    Kono Y (1978) Generation of superoxide radical during autoxidation of hydroxylamine and an assay for superoxide dismutase. Arch Biochem Biophys 186:189–195CrossRefGoogle Scholar
  36. 36.
    Luck H (1963) Catalase. In: Bergmeyer HW (ed) Methods of enzymatic analysis, section 3. Academic Press, New York, pp 885–894Google Scholar
  37. 37.
    Pearse AG (1960) This week’s citation classic. R Postgrad Mcd Sch 998Google Scholar
  38. 38.
    Hoesch RE, Koenig MA, Geocadin RG (2008) Coma after global ischemic brain injury: pathophysiology and emerging therapies. Crit Care Clin 24:25–44.  https://doi.org/10.1016/j.ccc.2007.11.003 CrossRefGoogle Scholar
  39. 39.
    Zhang X, Deguchi K, Yamashita T et al (2010) Temporal and spatial differences of multiple protein expression in the ischemic penumbra after transient MCAO in rats. Brain Res 1343:143–152.  https://doi.org/10.1016/j.brainres.2010.04.027 CrossRefGoogle Scholar
  40. 40.
    Li F, Irie K, Anwer MS, Fisher M (1997) Delayed triphenyltetrazolium chloride staining remains useful for evaluating cerebral infarct volume in a rat stroke model. J Cereb Blood Flow Metab 17:1132–1135.  https://doi.org/10.1097/00004647-199710000-00016 CrossRefGoogle Scholar
  41. 41.
    Adibhatla RM, Hatcher JF (2010) Lipid oxidation and peroxidation in CNS health and disease: from molecular mechanisms to therapeutic opportunities. Antioxid Redox Signal 12:125–169.  https://doi.org/10.1089/ars.2009.2668 CrossRefGoogle Scholar
  42. 42.
    Raghavendra Rao VL, Rao AM, Dogan A et al (2000) Glial glutamate transporter GLT-1 down-regulation precedes delayed neuronal death in gerbil hippocampus following transient global cerebral ischemia. Neurochem Int 36:531–537CrossRefGoogle Scholar
  43. 43.
    Bragin DE, Zhou B, Ramamoorthy P et al (2010) Differential changes of glutathione levels in astrocytes and neurons in ischemic brains by two-photon imaging. J Cereb Blood Flow Metab 30:734–738.  https://doi.org/10.1038/jcbfm.2010.9 CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Candelario-Jalil E, Mhadu NH, Al-Dalain SM et al (2001) Time course of oxidative damage in different brain regions following transient cerebral ischemia in gerbils. Neurosci Res 41:233–241CrossRefGoogle Scholar
  45. 45.
    Manto M, Bower JM, Conforto AB et al (2012) Consensus paper: roles of the cerebellum in motor control—the diversity of ideas on cerebellar involvement in movement. The Cerebellum 11:457–487.  https://doi.org/10.1007/s12311-011-0331-9 CrossRefGoogle Scholar
  46. 46.
    Hara A, Yoshimi N, Hirose Y et al (1995) DNA fragmentation in granular cells of human cerebellum following global ischemia. Brain Res 697:247–250CrossRefGoogle Scholar
  47. 47.
    Bello EP, Casas-Cordero R, Galiñanes GL et al (2016) Inducible ablation of dopamine D2 receptors in adult mice impairs locomotion, motor skill learning and leads to severe parkinsonism. Mol Psychiatry.  https://doi.org/10.1038/mp.2016.105 CrossRefGoogle Scholar
  48. 48.
    Martín A, Szczupak B, Gómez-Vallejo V et al (2013) PET imaging of serotoninergic neurotransmission with [11C]DASB and [18F]altanserin after focal cerebral ischemia in rats. J Cereb Blood Flow Metab 33:1967–1975.  https://doi.org/10.1038/jcbfm.2013.156 CrossRefPubMedPubMedCentralGoogle Scholar
  49. 49.
    Cao W, Drumheller A, Zaharia M et al (1993) Effects of experimentally induced ischemia on dopamine metabolism in rabbit retina. Invest Ophthalmol Vis Sci 34:3140–3146Google Scholar
  50. 50.
    Oliva I, Fernández M, Martín ED (2013) Dopamine release regulation by astrocytes during cerebral ischemia. Neurobiol Dis 58:231–241.  https://doi.org/10.1016/j.nbd.2013.06.007 CrossRefGoogle Scholar
  51. 51.
    Shimizu-Sasamata M, Yamamoto M, Okada M et al (1991) Effects of indeloxazine hydrochloride on behavioral and biochemical changes in the chronic phase of focal cerebral ischemia in rats. Arch Int Pharmacodyn Ther 314:74–89Google Scholar
  52. 52.
    Ferrari RS, Andrade CF, Ferrari RS, Andrade CF (2015) Oxidative stress and lung ischemia-reperfusion injury. Oxid Med Cell Longev 2015:1–14.  https://doi.org/10.1155/2015/590987 CrossRefGoogle Scholar
  53. 53.
    Stamenkovic S, Sekeljic V, Radenovic LAP (2012) Ischemia and neurogenesis: link between neurodegeneration and repair. In: ICG, AW (eds) Neurogenesis research: new developments. NOVA Science Publishers, Inc. New York, pp 115–136Google Scholar
  54. 54.
    Lichtenwalner RJ, Parent JM (2006) Adult neurogenesis and the ischemic forebrain. J Cereb Blood Flow Metab 26:1–20.  https://doi.org/10.1038/sj.jcbfm.9600170 CrossRefPubMedGoogle Scholar
  55. 55.
    Vandenbosch R, Borgs L, Beukelaers P et al (2009) Adult neurogenesis and the diseased brain. Curr Med Chem 16:652–666CrossRefGoogle Scholar
  56. 56.
    Ziemka-Nałęcz M, Zalewska T (2012) Endogenous neurogenesis induced by ischemic brain injury or neurodegenerative diseases in adults. Acta Neurobiol Exp (Wars) 72:309–324Google Scholar
  57. 57.
    Chen A, Xiong L-J, Tong Y, Mao M (2013) The neuroprotective roles of BDNF in hypoxic ischemic brain injury. Biomed Rep 1:167–176.  https://doi.org/10.3892/br.2012.48 CrossRefGoogle Scholar
  58. 58.
    Ploughman M, Windle V, MacLellan CL et al (2009) Brain-derived neurotrophic factor contributes to recovery of skilled reaching after focal ischemia in rats. Stroke 40:1490–1495.  https://doi.org/10.1161/STROKEAHA.108.531806 CrossRefGoogle Scholar
  59. 59.
    Xie Z-M, Wang X-M, Xu N et al (2017) Alterations in the inflammatory cytokines and brain-derived neurotrophic factor contribute to depression-like phenotype after spared nerve injury: improvement by ketamine. Sci Rep 7:3124.  https://doi.org/10.1038/s41598-017-03590-3 CrossRefPubMedPubMedCentralGoogle Scholar

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© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of PhysiologyArmed Forces Medical CollegePuneIndia
  2. 2.Department of BiophysicsPanjab UniversityChandigarhIndia
  3. 3.Department of BiochemistryPanjab UniversityChandigarhIndia

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