Abstract
Neonatal stroke occurs in one in 4,000 live births and leads to significant morbidity and mortality. Approximately two thirds of the survivors have long-term sequelae including seizures and neurological deficits. However, the pathophysiological mechanisms of recovery after neonatal stroke are not clearly understood, and preventive measures and treatments are nonexistent in the clinical setting. In this study, we investigated the effect of vascular endothelial growth factor (VEGF) treatment on histological recovery and angiogenic response to the developing brain after an ischemic insult. Ten-day-old Sprague–Dawley rats underwent right middle cerebral arterial occlusion (MCAO) for 1.5 h. Diffusion-weighted MRI during occlusion confirmed focal ischemia that was then followed by reperfusion. On group of animals received 5-bromo-2-deoxyuridine and sacrificed at postnatal day (P)18 or P25. A second group of animals was treated with VEGF (1.5 μg/kg, icv) or phosphate-buffered saline (PBS) at P18 and perfusion fixed at P25. Based on Nissl and iron staining, a single VEGF injection reduced the injury score, compared to the animals that underwent MCAO and PBS injection. Furthermore, neurodegeneration represented by neuronal nuclei staining was markedly diminished. In addition, animals treated with VEGF revealed a positive trend in endothelial proliferation and a significant increase in total vessel volume in the peri-infarct region of the caudate. The number of Iba1-positive microglial cells was significantly reduced after a single VEGF injection, and myelin basic protein expression was enhanced in the caudate after ischemia without an effect of VEGF treatment. In conclusion, delayed treatment with VEGF ameliorates injury, promotes endothelial cell proliferation, and increases total vascular volume following neonatal stroke. These results suggest that VEGF has a neuroprotective effect, in part by enhancing endogenous angiogenesis. These data contribute to a better understanding of neonatal stroke.
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References
Nelson KB. Perinatal ischemic stroke. Stroke. 2007;38:742–5.
Sreenan C, Bhargava R, Robertson CMT. Cerebral infarction in the term newborn: clinical presentation and long-term outcome. J Pediatr. 2000;137:351–5.
Ferriero DM. Neonatal brain injury. NEJM. 2004;351:1985–95.
Arai K, Jin G, Navaratna D, Lo EH. Brain angiogenesis in developmental and pathological processes: neurovascular injury and angiogenic recovery after stroke. FEBS J. 2009;276:4644–52.
Ohab JJ, Fleming S, Blesch A, Carmichael ST. A neurovascular niche for neurogenesis after stroke. J Neurosci. 2006;26:13007–16.
Madri J. Modeling the neurovascular niche: implications for recovery from CNS injury. J Physiol Pharmacol. 2009;60:94–104.
Hayashi T, Iwai M, Ikeda T, Jin G, Deguchi K, Nagotani S, et al. Neural precursor cells division and migration in neonatal rat brain after ischemic/hypoxic injury. Brain Res. 2005;1038:41–9.
Spadafora R, Gonzalez FF, Derugin N, Wendland M, Ferriero D, McQuillen P. Altered fate of subventricular zone progenitor cells and reduced neurogenesis following neonatal stroke. Dev Neurosci. 2010;32:101–13.
Zhang ZG. VEGF enhances angiogenesis and promotes blood–brain barrier leakage in the ischemic brain. J Clin Invest. 2000;106:829–38.
Derugin N, Wendland M, Muramatsu K, Roberts TPL, Gregory G, Ferriero DM, et al. Evolution of brain injury after transient middle cerebral artery occlusion in neonatal rats editorial comment. Stroke. 2000;31:1752–61.
Mu D. Regulation of hypoxia-inducible factor 1alpha and induction of vascular endothelial growth factor in a rat neonatal stroke model. Neurobiol Dis. 2003;14:524–34.
Dobbing J, Sands J. Comparative aspects of the brain growth spurt. Early Hum Dev. 1979;3:79–83.
Derugin N, Ferriero DM, Vexler ZS. Neonatal reversible focal cerebral ischemia: a new model. Neurosci Res. 1998;32:349–53.
Derugin N, Dingman A, Wendland MF, Fox C, Bollen A, Vexler ZS. Magnetic resonance imaging as a surrogate measure for histological sub-chronic endpoint in a neonatal rat stroke model. Brain Res. 2005;1066:49–56.
Shimotake J, Derugin N, Wendland M, Vexler ZS, Ferriero DM. Vascular endothelial growth factor receptor-2 inhibition promotes cell death and limits endothelial cell proliferation in a neonatal rodent model of stroke. Stroke. 2010;41:343–9.
Paxinos G. The rat brain in stereotaxic coordinates. 4th ed. San Diego: Academic; 1998.
Faustino JV, Wang X, Johnson CE, Klibanov A, Derugin N, Wendland MF, et al. Microglial cells contribute to endogenous brain defenses after acute neonatal focal stroke. J Neurosci. 2011;31:12992–3001.
Greenberg DA, Jin K. From angiogenesis to neuropathology. Nature. 2005;438:954–9.
Kaya D, Gürsoy-Özdemir Y, Yemisci M, Tuncer N, Aktan S, Dalkara T. VEGF protects brain against focal ischemia without increasing blood–brain permeability when administered intracerebroventricularly. J Cereb Blood Flow Metab. 2005;25:1111–8.
Sun Y. VEGF-induced neuroprotection, neurogenesis, and angiogenesis after focal cerebral ischemia. J Clin Invest. 2003;111:1843–51.
Wang Y, Galvan V, Gorostiza O, Ataie M, Jin K, Greenberg DA. Vascular endothelial growth factor improves recovery of sensorimotor and cognitive deficits after focal cerebral ischemia in the rat. Brain Res. 2006;1115:186–93.
Wang Y-Q, Cui H-R, Yang S-Z, Sun H-P, Qiu M-H, Feng X-Y, et al. VEGF enhance cortical newborn neurons and their neurite development in adult rat brain after cerebral ischemia. Neurochem Int. 2009;55:629–36.
Chen J, Chopp M. Neurorestorative treatment of stroke: cell and pharmacological approaches. J Am Soc Exp NeuroTher. 2011;3:466–73.
Chopp M, Li Y, Zhang J. Plasticity and remodeling of brain. J Neurol Sci. 2008;265:97–101.
Krupinski J, Stroemer P, Slevin M, Marti E, Kumar P, Rubio F. Three-dimensional structure and survival of newly formed blood vessels after focal cerebral ischemia. NeuroReport. 2003;14:1171–6.
Ma Y, Zechariah A, Qu Y, Hermann DM. Effects of vascular endothelial growth factor in ischemic stroke. J Neurosci Res. 2012. doi:10.1002/jnr.23088.
Hermann DM, Zechariah A. Implications of vascular endothelial growth factor for postischemic neurovascular remodeling. J Cereb Blood Flow Metab. 2009;29:1620–43.
Sc C, Chopp M. Recovery recapitulates ontogeny. Trends Neurosci. 2000;23:265–71.
Dellian M, Witwer BP, Salehi HA, Yuan F, Jain RK. Quantitation and physiological characterization of angiogenic vessels in mice. Am J Pathol. 1996;149:59–71.
Zhang ZG, Chopp M. Vascular endothelial growth factor and angiopoietins in focal cerebral ischemia. Trends Cardiovasc Med. 2002;12:62–6.
Denker SP, Ji S, Dingman A, Lee SY, Derugin N, Wendland MF, et al. Macrophages are comprised of resident brain microglia not infiltrating peripheral monocytes acutely after neonatal stroke. J Neurochem. 2007;100:893–904.
Acker T, Beck H, Plate KH. Cell type specific expression of vascular endothelial growth factor and angiopoietin-1 and −2 suggests an important role of astrocytes in cerebellar vascularization. Mech Dev. 2001;108:45–7.
Salhia B, Angelova L, Roncari L, Wu X, Shannon P, Guha A. Expression of vascular endothelial growth factor by reactive astrocytes and associated neoangiogenesis. Brain Res. 2000;883:1–11.
Proescholdt M, Heiss J, Walbridge S, Mühlhauser J, Capogrossi M, Oldfield E, et al. Vascular endothelial growth factor (VEGF) modulates vascular permeability and inflammation in rat brain. J Neuropathol Exp Neurol. 1999;58:613–27.
Horie N, Pereira MP, Niizuma K, Sun G, Keren-Gill H, Encarnacion A, et al. Transplanted stem cell-secreted vascular endothelial growth factor effects poststroke recovery, inflammation, and vascular repair. Stem Cells. 2011;29:274–85.
Olivier AK, Gallup JM, Geelen AV, Ackermann MR. Exogenous administration of vascular endothelial growth factor prior to human respiratory syncytial virus a2 infection reduces pulmonary pathology in neonatal lambs and alters epithelial innate immune responses. Exp Lung Res. 2011;37:131–43.
Fernandez-Lopez D, Faustino J, Daneman R, Zhou L, Lee SY, Derugin N, et al. Blood–brain barrier permeability is increased after acute adult stroke but not neonatal stroke in the rat. J Neurosci. 2012;32:9588–600.
Li S, Overman JJ, Katsman D, Kozlov SV, Donnelly CJ, Twiss JL, et al. An age-related sprouting transcriptome provides molecular control of axonal sprouting after stroke. Nat Neurosci. 2010;13:1496–504.
Liu Z, Li Y, Zhang ZG, Cui X, Cui Y, Lu M, et al. Bone marrow stromal cells enhance inter- and intracortical axonal connections after ischemic stroke in adult rats. J Cereb Blood Flow Metab. 2010;30:1288–95.
Ueno Y, Chopp M, Zhang L, Buller B, Liu Z, Lehman NL, et al. Axonal outgrowth and dendritic plasticity in the cortical peri-infarct area after experimental stroke. Stroke. 2012. doi:10.1161/STROKEAHA.111.646224/-/DC1.
Hayakawa K, Seo JH, Pham L-DD, Miyamoto N, Som AT, Guo S, et al. Cerebral endothelial derived vascular endothelial growth factor promotes the migration but not the proliferation of oligodendrocyte precursor cells in vitro. Neurosci Lett. 2012;513:42–6.
Hayakawa K, Pham LDD, Som AT, Lee BJ, Guo S, Lo EH, et al. Vascular endothelial growth factor regulates the migration of oligodendrocyte precursor cells. J Neurosci. 2011;31:10666–70.
Kaur C, Sivakumar V, Ang LS, Sundaresan A. Hypoxic damage to the periventricular white matter in neonatal brain: role of vascular endothelial growth factor, nitric oxide and excitotoxicity. J Neurochem. 2006;98:1200–16.
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This work was supported by a grant NS35902 (DMF and ZSV) and was a recipient of PAS Travel Award 2012 (MD).
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ZS Vexler and DM Ferriero are co-senior authors.
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Dzietko, M., Derugin, N., Wendland, M.F. et al. Delayed VEGF Treatment Enhances Angiogenesis and Recovery After Neonatal Focal Rodent Stroke. Transl. Stroke Res. 4, 189–200 (2013). https://doi.org/10.1007/s12975-012-0221-6
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DOI: https://doi.org/10.1007/s12975-012-0221-6