En trombotisk hjerneslag modell basert på Transient Cerebral Hypoksi-iskemi

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Sun, Y. Y., Kuan, C. Y. A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia. J. Vis. Exp. (102), e52978, doi:10.3791/52978 (2015).

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Abstract

Stroke forskning har tålt mange tilbakeslag i å oversette nevro terapi i klinisk praksis. I motsetning til den virkelige verden terapi (Pa trombolyse) produserer sjelden fordeler i mekaniske okklusjon-baserte eksperimentelle modeller, som dominerer preklinisk hjerneslag forskning. Denne fordelingen mellom benk og nattbord antyder behovet for å ansette Pa-responsive modeller i preklinisk hjerneslag forskning. For dette formål, er en enkel og tPA-reaktive trombotisk slag modell oppfunnet og beskrevet her. Denne modellen består av transient okklusjon av den ensidige arteria carotis communis og levering av 7,5% oksygen gjennom en ansiktsmaske i voksen mus i 30 minutter, under opprettholdelse av dyret rektal temperatur på 37,5 ± 0,5 ° C. Selv reversibel ligering av den ensidige carotid arterie eller hypoksi hver trykkes cerebral blodstrøm bare forbigående, er kombinasjonen av begge fornærmelser forårsaket varig reperfusjon underskudd, fibrin og blodplateavleiring, og store INFARct i midten cerebral arterie-leverte territorium. Viktigere, haleveneinjeksjon av rekombinant tPA ved 0,5, 1, eller fire timer etter Thi (10 mg / kg) gitt tidsavhengig reduksjon av dødelighet og infarktstørrelsen. Denne nye slagmodell, er enkel og kan standardiseres tvers laboratorier for å sammenligne eksperimentelle resultater. Videre induserer det trombose uten craniectomy eller innføre forhånds dannet emboli. Gitt disse unike fordeler, er thi modellen et nyttig tillegg til repertoaret av preklinisk hjerneslag forskning.

Materials

Name Company Catalog Number Comments
adult male mice Charles River C57BL/6  10-13 weeks old (22-30 g)
Mobile Laboratory Animal Anesthesia System VetEquip 901807 anesthesia
Medical air (Compressed) air tank Airgas UN1002 anesthesia
Isoflurane Piramal Healthcare NDC 66794-013-25 anesthesia
Multi-Station Lab Animal AnesthesiaSystem Surgivet V703501 hypoxia system
7.5% O2 balanced by 92.5% N2 tank Airgas UN1956 hypoxia system
Temperature Controller with heating lamp  Cole Parmer  EW-89000-10 temperature controllers
Rectal probe Cole Parmer  NCI-00141PG temperature controllers
Dissecting microscope  Olympus  SZ40 surgical setup
Heat pump with warming pad Gaymar  TP700 surgical setup
Fine curved forceps (serrated) FST 11370-31 surgical instrument
Fine curved forceps (smooth) FST 11373-12 surgical instrument
micro scissors FST 15000-03 surgical instrument
micro needle holders FST 12060-01 surgical instrument
Halsted-Mosquito hemostats FST 13008-12 surgical instrument
5-0 silk suture  Harvard Apparatus 624143 surgical supplies
4-0 Nylon monofilament suture LOOK 766B surgical supplies
Tissue glue Abbott Laboratories NC9855218 surgical supplies
Puralube Vet ointment Fisher NC0138063  eye dryness prevention 
MoorFLPI-2 blood flow imager Moor 780-nm laser source Laser Speckle Contrast Imaging
Mannitol Sigma M4125 in vivo TTC
2,3,5-triphenyltetrazolium chloride (TTC)  Sigma T8877 in vivo TTC
Vibratome Stoelting 51425 brain section for in vivo TTC 
Digital microscope Dino-Lite AM2111 whole-brain imaging
O.C.T compound Sakura Finetek 4583
goat anti-rabbit Alexa Fluro 488 Invitrogen A11008 Immunohistochemistry
Cryostat Vibratome ultrapro 5000 brain section for IHC
Evans blue Sigma E2129 Detecting vascular perfusion
Microtome Electron Microscopy Sciences 5000 brain section for histology
Avertin (2, 2, 2-Tribromoethanol) Sigma T48402 euthanasia
Fluorescent microscope Olympus DP73
Meloxicam SR ZooPharm NSAID analgesia

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References

  1. Broderick, J. P., Hacke, W. Treatment of acute ischemic stroke: Part I: recanalization strategies. Circulation. 106, (12), 1563-1569 (2002).
  2. Hossmann, K. A. Pathophysiological basis of translational stroke research. Folia Neuropathol. 47, (3), 213-227 (2009).
  3. Hossmann, K. A. The two pathophysiologies of focal brain ischemia: implications for translational stroke research. J. Cereb. Blood Flow Metab. 32, (7), 1310-1316 (2012).
  4. Macrae, I. M. Preclinical stroke research--advantages and disadvantages of the most common rodent models of focal ischaemia. Br. J. Pharmacol. 164, (4), 1062-1078 (2011).
  5. Niessen, F., Hilger, T., Hoehn, M., Hossmann, K. A. Differences in clot preparation determine outcome of recombinant tissue plasminogen activator treatment in experimental thromboembolic stroke. Stroke. 34, (8), 2019-2024 (2003).
  6. Orset, C., et al. Mouse model of in situ thromboembolic stroke and reperfusion. Stroke. 38, (10), 2771-2778 (2007).
  7. Watson, B. D., Dietrich, W. D., Prado, R., Ginsberg, M. D. Argon laser-induced arterial photothrombosis. Characterization and possible application to therapy of arteriovenous malformations. J. Neurosurgery. 66, (5), 748-754 (1987).
  8. Sun, Y. Y., et al. Synergy of combined tPA-edaravone therapy in experimental thrombotic stroke. PLoS One. 9, e98807 (2014).
  9. Levine, S. Anoxic-ischemic encephalopathy in rats. Am. J. Pathol. 36, 1-17 (1960).
  10. Rice, J. E. 3rd, Vannucci, R. C., Brierley, J. B. The influence of immaturity on hypoxic-ischemic brain damage in the rat. Annals Neurol. 9, (2), 131-141 (1981).
  11. Vannucci, S. J., et al. Experimental stroke in the female diabetic, db/db, mouse. J. Cereb. Blood Flow Metab. 21, (2), 52-60 (2001).
  12. Adhami, F., et al. Cerebral ischemia-hypoxia induces intravascular coagulation and autophagy. Am. J. Pathol. 169, (2), 566-583 (2006).
  13. Shereen, A., et al. Ex vivo diffusion tensor imaging and neuropathological correlation in a murine model of hypoxia-ischemia-induced thrombotic stroke. J. Cereb. Blood Flow Metab. 31, (4), 1155-1169 (2011).
  14. Michaud, J. P., Pimentel-Coelho, P. M., Tremblay, Y., Rivest, S. The impact of Ly6C low monocytes after cerebral hypoxia-ischemia in adult mice. J. Cereb. Blood Flow Metab. 34, (7), e1-e9 (2014).
  15. Zoppo, G. J. Virchow's triad: the vascular basis of cerebral injury. Rev. Neurol. Dis. 5, 12-21 (2008).
  16. Dunn, A. K. Laser speckle contrast imaging of cerebral blood flow. Annals Biomed. Eng. 40, (2), 367-377 (2012).
  17. Sun, Y. Y., Yang, D., Kuan, C. Y. Mannitol-facilitated perfusion staining with 2,3,5-triphenyltetrazolium chloride (TTC) for detection of experimental cerebral infarction and biochemical analysis. J. Neurosci. Methods. 203, (1), 122-129 (2012).
  18. Lozano, R., et al. Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: a systematic analysis for the Global Burden of Disease Study. Lancet. 380, (9859), 2095-2128 (2010).
  19. Murray, C. J., et al. 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. Lancet. 380, (9859), 2197-2223 (2012).
  20. Dirnagl, U., Macleod, M. R. Stroke research at a road block: the streets from adversity should be paved with meta-analysis and good laboratory practice. Br. J. Pharm. 157, (7), 1154-1156 (2009).
  21. Dirnagl, U., et al. A concerted appeal for international cooperation in preclinical stroke research. Stroke. 44, (6), 1754-1760 (2013).
  22. Khatri, P., et al. Revascularization end points in stroke interventional trials: recanalization versus reperfusion in IMS-I. Stroke. 36, (11), 2400-2403 (2005).
  23. Rosenberg, R. D., Aird, W. C. Vascular-bed--specific hemostasis and hypercoagulable states. New Eng. J. Med. 340, (20), 1555-1564 (1999).
  24. Majid, A., et al. Differences in vulnerability to permanent focal cerebral ischemia among 3 common mouse strains. Stroke. 31, (11), 2707-2714 (2000).

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