Abstract
Ischemic stroke is one of the major causes of devastating neurological disabilities and mortality worldwide. Despite extensive research for treatment approaches, there remains limited therapy in the stroke field. Therefore, more research is required for reproducibility to understand stroke pathology in pre-clinical studies. In the current modified method, mice were subjected to photothrombotic stroke (pt-MCA; proximal-middle cerebral artery was exposed with a 532 nm laser beam for 4 min) by retro-orbital injection of photosensitive dye, Rose Bengal (15 mg/kg) before the laser light exposure. Sensorimotor deficits were assessed by rotarod and catwalk test at 72 h following post-pt-MCAO, and brain samples were collected for infarct volume and hemorrhagic transformation (HT) assessments. Cognitive impairments were assessed by a novel objective recognition and Morris’s water maze tests at the end of the follow-up. pt-MCAO animals significantly reduced body weight and impaired motor and cognitive functions. Furthermore, pt-MCAO animals showed apparent infarction, brain edema, and increased HT compared to the sham animals. Additionally, this method enables concurrent measurement of short-term and long-term neurological dysfunction with relatively larger cortical and sub-cortical infarct volume following pt-MCAO. With respect to the other models, this modified model offers enhanced reproducibility regarding infarct volume and cognitive/functional outcomes and avoids complications associated with critical surgeries and craniotomy. In conclusion, this modified model helps to understand stroke pathogenesis and minimize the animals’ numbers which help to increase the scientific and statistical potential in pre-clinical studies.
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The data generated and analyzed in this study will be available from the corresponding author upon reasonable request.
References
Ahmad S, Pandya C, Kindelin A et al (2020) C3a receptor antagonist therapy is protective with or without thrombolysis in murine thromboembolic stroke. Br J Pharmacol 177:2466–2477. https://doi.org/10.1111/bph.14989
Ahmed HA, Ismael S, Salman M et al (2022) Direct AT2R stimulation slows post-stroke cognitive decline in the 5XFAD Alzheimer’s disease mice. Mol Neurobiol 59:4124–4140. https://doi.org/10.1007/s12035-022-02839-x
Allen CL, Bayraktutan U (2009) Oxidative stress and Its role in the pathogenesis of Ischaemic stroke. Int J Stroke 4:461–470. https://doi.org/10.1111/j.1747-4949.2009.00387.x
Chen ST, Hsu CY, Hogan EL et al (1986) A model of focal ischemic stroke in the rat: reproducible extensive cortical infarction. Stroke 17:738–743. https://doi.org/10.1161/01.STR.17.4.738
Dahlqvist P, Ronnback A, Bergstrom S-A et al (2004) Environmental enrichment reverses learning impairment in the Morris water maze after focal cerebral ischemia in rats. Eur J Neurosci 19:2288–2298. https://doi.org/10.1111/j.0953-816X.2004.03248.x
Dietrich WD, Busto R, Watson BD et al (1987a) Photochemically induced cerebral infarction. Acta Neuropathol 72:326–334. https://doi.org/10.1007/BF00687263
Dietrich WD, Watson BD, Busto R et al (1987b) Photochemically induced cerebral infarction. Acta Neuropathol 72:315–325. https://doi.org/10.1007/BF00687262
Feigin VL, Norrving B, Mensah GA (2017) Global Burden of Stroke. Circ Res 120:439–448. https://doi.org/10.1161/CIRCRESAHA.116.308413
Feigin VL, Stark BA, Johnson CO et al (2021) Global, regional, and national burden of stroke and its risk factors, 1990–2019: a systematic analysis for the Global Burden of disease study 2019. Lancet Neurol 20:795–820. https://doi.org/10.1016/S1474-4422(21)00252-0
Fonarow GC, Smith EE, Saver JL et al (2011) Timeliness of tissue-type plasminogen activator therapy in acute ischemic stroke. Circulation 123:750–758. https://doi.org/10.1161/CIRCULATIONAHA.110.974675
Ismael S, Nasoohi S, Yoo A et al (2020) Tissue plasminogen activator promotes TXNIP-NLRP3 Inflammasome activation after Hyperglycemic stroke in Mice. Mol Neurobiol 57:2495–2508. https://doi.org/10.1007/s12035-020-01893-7
Ismael S, Patrick D, Salman M et al (2022) Verapamil inhibits TXNIP-NLRP3 inflammasome activation and preserves functional recovery after intracerebral hemorrhage in mice. Neurochem Int. https://doi.org/10.1016/j.neuint.2022.105423
Ismael S, Zhao L, Nasoohi S, Ishrat T (2018) Inhibition of the NLRP3-inflammasome as a potential approach for neuroprotection after stroke. Sci Rep 8:5971. https://doi.org/10.1038/s41598-018-24350-x
Lansberg MG, Bluhmki E, Thijs VN (2009) Efficacy and safety of tissue plasminogen activator 3 to 4.5 hours after acute ischemic stroke. Stroke 40:2438–2441. https://doi.org/10.1161/STROKEAHA.109.552547
Li Y, Zhang J (2021) Animal models of stroke. Anim Model Exp Med 4:204–219. https://doi.org/10.1002/ame2.12179
Lin X, Wang H, Chen J et al (2022) Nonhuman primate models of ischemic stroke and neurological evaluation after stroke. J Neurosci Methods. https://doi.org/10.1016/j.jneumeth.2022.109611
Llovera G, Hofmann K, Roth S et al (2015) Results of a preclinical randomized controlled multicenter trial (pRCT): Anti-CD49d treatment for acute brain ischemia. Sci Transl Med. https://doi.org/10.1126/scitranslmed.aaa9853
Llovera G, Roth S, Plesnila N et al (2014) Modeling stroke in mice: permanent coagulation of the distal middle cerebral artery. J vis Exp. https://doi.org/10.3791/51729
Morris RGM, Garrud P, Rawlins JNP, O’Keefe J (1982) Place navigation impaired in rats with hippocampal lesions. Nature 297:681–683. https://doi.org/10.1038/297681a0
Murray CJL, Vos T, Lozano R et al (2012) Disability-adjusted life years (DALYs) for 291 diseases and injuries in 21 regions, 1990–2010: a systematic analysis for the Global Burden of Disease Study 2010. Lancet 380:2197–2223. https://doi.org/10.1016/S0140-6736(12)61689-4
Puurunen K, Jolkkonen J, Sirviö J et al (2001) An α2-adrenergic antagonist, atipamezole, facilitates behavioral recovery after focal cerebral ischemia in rats. Neuropharmacology 40:597–606. https://doi.org/10.1016/S0028-3908(00)00182-9
Rosenblum WI, El-Sabban F (1977) Platelet aggregation in the cerebral microcirculation: effect of aspirin and other agents. Circ Res 40:320–328. https://doi.org/10.1161/01.RES.40.3.320
Salman M, Ismael S, Li L et al (2022) Acute hyperglycemia exacerbates hemorrhagic transformation after embolic stroke and reperfusion with tPA: a possible role of TXNIP-NLRP3 inflammasome. J Stroke Cerebrovasc Dis. https://doi.org/10.1016/j.jstrokecerebrovasdis.2021.106226
Salman M, Ismael S, Li L et al (2021) Endothelial thioredoxin-interacting protein depletion reduces Hemorrhagic transformation in Hyperglycemic mice after embolic stroke and thrombolytic therapy. Pharmaceuticals 14:983. https://doi.org/10.3390/ph14100983
Salman M, Tabassum H, Parvez S (2020) Nrf2/HO-1 mediates neuroprotective effects of pramipexole by attenuating oxidative damage and mitochondrial perturbation after traumatic brain injury. Dis Model Mech. https://doi.org/10.1242/dmm.045021
Türeyen K, Vemuganti R, Sailor KA, Dempsey RJ (2004) Infarct volume quantification in mouse focal cerebral ischemia: a comparison of triphenyltetrazolium chloride and cresyl violet staining techniques. J Neurosci Methods 139:203–207. https://doi.org/10.1016/j.jneumeth.2004.04.029
Watson BD, Dietrich WD, Busto R et al (1985) Induction of reproducible brain infarction by photochemically initiated thrombosis. Ann Neurol 17:497–504. https://doi.org/10.1002/ana.410170513
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
Yang L, Shah KK, Abbruscato TJ (2012) An In Vitro Model of Ischemic Stroke. pp 451–466
Acknowledgements
This work was supported by the National Institutes of Health under the award number R56NS127924-01 and the UTHSC Bridge funding award E073005058-Bridge Support-2022 to Dr T. Ishrat and American Heart Association (AHA) Postdoctoral Fellowship 1029163 to Dr M. Salman.
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M.S: Conceptualization, Methodology, Investigation, Validation, Data curation, Formal analysis, Writing–original draft preparation. S.I: Conceptualization, Methodology, Writing–review and editing. T.I: Conceptualization, Supervision, Funding acquisition, Writing–review and editing.
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All procedures related to animal studies were approved by the institutional animal committee at UTHSC in full accordance with the ethical guidelines of the National Institutes of Health for the care and use of laboratory animals.
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Communicated by Sreedharan Sajikumar.
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Salman, M., Ismael, S. & Ishrat, T. A modified murine photothrombotic stroke model: a minimally invasive and reproducible cortical and sub-cortical infarct volume and long-term deficits. Exp Brain Res 241, 2487–2497 (2023). https://doi.org/10.1007/s00221-023-06696-5
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DOI: https://doi.org/10.1007/s00221-023-06696-5