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Neuroscience

Analysis of Cerebral Vasospasm in a Murine Model of Subarachnoid Hemorrhage with High Frequency Transcranial Duplex Ultrasound

Published: June 3rd, 2021

DOI:

10.3791/62186

1Department of Neurosurgery, University Medical Center of the Johannes Gutenberg-University of Mainz, 2Center for Cardiology - Cardiology I, University Medical Center of the Johannes Gutenberg-University of Mainz, 3Center for Thrombosis and Hemostasis (CTH), University Medical Center of the Johannes Gutenberg-University of Mainz, 4German Center for Cardiovascular Research (DZHK) - Partner site Rhine-Main, 5Department of Anesthesiology, University Medical Center of the Johannes Gutenberg-University of Mainz
* These authors contributed equally

The aim of this manuscript is to present a sonography-based method that allows in vivo imaging of blood flow in cerebral arteries in mice. We demonstrate its application to determine changes in blood flow velocities associated with vasospasm in murine models of subarachnoid hemorrhage (SAH).

Cerebral vasospasm that occurs in the weeks after subarachnoid hemorrhage, a type of hemorrhagic stroke, contributes to delayed cerebral ischemia. A problem encountered in experimental studies using murine models of SAH is that methods for in vivo monitoring of cerebral vasospasm in mice are lacking. Here, we demonstrate the application of high frequency ultrasound to perform transcranial Duplex sonography examinations on mice. Using the method, the internal carotid arteries (ICA) could be identified. The blood flow velocities in the intracranial ICAs were accelerated significantly after induction of SAH, while blood flow velocities in the extracranial ICAs remained low, indicating cerebral vasospasm. In conclusion, the method demonstrated here allows functional, noninvasive in vivo monitoring of cerebral vasospasm in a murine SAH model.

Spontaneous subarachnoid hemorrhage (SAH) is a form of hemorrhagic stroke mostly caused by the rupture of an intracranial aneurysm1. The neurological outcome is mainly influenced by two factors: early brain injury (EBI), which is caused by the effects of the bleeding and the associated transient global cerebral ischemia, and delayed cerebral ischemia (DCI), which occurs during the weeks following the bleeding2,3. DCI was reported to affect up to 30% of SAH patients2. The pathophysiology of DCI involves angiographic cerebral vasospasm, a disturbed microcirculation....

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The animal experiments were approved by the responsible animal care committee (Landesuntersuchungsamt Rheinland-Pfalz) and conducted in accordance with the German Animal Welfare Act (TierSchG). All applicable international, national, and institutional guidelines for the care and use of animals were followed. In this study, we performed measurements of blood flow velocities of intracranial and extracranial arteries in female C57BL/6N mice aged 11-12 weeks with a body weight between 19-21 g. The mice were subjected to eith.......

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In 6 mice, in 3 of which SAH was induced using the endovascular filament perforation model while 3 obtained sham surgery, the blood flow velocities of the intracranial internal carotid artery (ICA) and of the extracranial ICA were determined one day before surgery, and 1, 3, and 7 days after surgery. The measurements were performed as part of the echocardiography examinations of another study under anesthesia with isoflurane while maintaining the body temperature at 37 °C19.

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To the best of our knowledge, this study is the first to present a protocol for monitoring of cerebral vasospasm in a murine model of SAH with high frequency transcranial color-coded Duplex ultrasound. We show that this method can measure an increase in intracranial blood flow velocities after SAH induction in mice. In human medicine this phenomenon is well known3,15. Several clinical studies have shown that elevated blood flow velocities of the large intracrania.......

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The authors would like to thank Stefan Kindel for preparation of the illustrations in the video. PW, MM and SHK were supported by the German Federal Ministry for Education and Research (BMBF 01EO1503). The work was supported by a Large Instrumentation Grant of the German Research Foundation (DFG INST 371/47-1 FUGG). MM was supported by a grant from the Else Kröner-Fresenius-Stiftung (2020_EKEA.144).

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Name Company Catalog Number Comments
Balea hair removal creme Balea; Germany ASIN B0759XM39V hair removal creme
C57BL/6N mice Janvier; Saint-Berthevin Cedex, France n.a. mice
Corneregel Bausch&Lomb; Rochester, NY, USA REF 81552983 eye ointment, lube
cotton swabs Hecht Assistent; Sondenheim vor der Röhn, Germany REF 44302010 cotton swabs
Ecco-XS razor Tondeo; Soligen, Germany DE 28693396 razor
Electrode cream GE; Boston, MA, USA REF 21708318 conductive paste
Heating plate Medax; Kiel, Germany 2005-205-01
Isoflurane Abvie; Wiesbaden, Germany n.a. volatile anesthetic
Leukofix BSN medical; Hamburg, Germany REF 02137-00 tape
Mechanical arm + micromanipulator VisualSonics; FujiFilm, Toronto, CA P/N 11277
Microbac tissues Paul Hartmann AG; Hamburg, Germany REF 981387 antimicrobial tissues
MZ400, 38 MHz linear array transducer VisualSonics; FujiFilm, Toronto, CA REF 51068-30 ultrasound transducer
Sonosid ASID Bonz GmbH; Herrenberg, Germany REF 782010 ultrasonography gel
Ultrasound platform with heating plate and ECG-recording VisualSonics; FujiFilm, Toronto, CA P/N 11179
UniVet-Porta Groppler; Oberperasberg, Germany S/N BKGM0437 isoflurane vaporizer
Vevo3100 VisualSonics; FujiFilm, Toronto, CA REF 51073-45 ultrasonography device
VevoLab software VisualSonics; FujiFilm, Toronto, CA n.a. evaluation software

  1. Macdonald, R. L., Schweizer, T. A. Spontaneous subarachnoid haemorrhage. Lancet. 389 (10069), 655-666 (2017).
  2. Macdonald, R. L. Delayed neurological deterioration after subarachnoid haemorrhage. Nature Reviews Neurology. 10 (1), 44-58 (2014).
  3. Francoeur, C. L., Mayer, S. A. Management of delayed cerebral ischemia after subarachnoid hemorrhage. Critical Care. 20 (1), 277 (2016).
  4. van Lieshout, J. H., et al. An introduction to the pathophysiology of aneurysmal subarachnoid hemorrhage. Neurosurgical Review. , (2017).
  5. Altay, T., et al. A novel method for subarachnoid hemorrhage to induce vasospasm in mice. J Neurosci Methods. 183 (2), 136-140 (2009).
  6. Momin, E. N., et al. Controlled delivery of nitric oxide inhibits leukocyte migration and prevents vasospasm in haptoglobin 2-2 mice after subarachnoid hemorrhage. Neurosurgery. 65 (5), 937-945 (2009).
  7. Froehler, M. T., et al. Vasospasm after subarachnoid hemorrhage in haptoglobin 2-2 mice can be prevented with a glutathione peroxidase mimetic. Journal of Clinical Neuroscience. 17 (9), 1169-1172 (2010).
  8. Provencio, J. J., Altay, T., Smithason, S., Moore, S. K., Ransohoff, R. M. Depletion of Ly6G/C(+) cells ameliorates delayed cerebral vasospasm in subarachnoid hemorrhage. Journal of Neuroimmunology. 232 (1-2), 94-100 (2011).
  9. Kamp, M. A., et al. Evaluation of a murine single-blood-injection SAH model. PLoS One. 9 (12), 114946 (2014).
  10. Luh, C., et al. The Contractile Apparatus Is Essential for the Integrity of the Blood-Brain Barrier After Experimental Subarachnoid Hemorrhage. Translational Stroke Research. , (2018).
  11. Neulen, A., et al. A Volumetric Method for Quantification of Cerebral Vasospasm in a Murine Model of Subarachnoid Hemorrhage. Journal of Visualized Experiments. (137), (2018).
  12. Neulen, A., et al. Large Vessel Vasospasm Is Not Associated with Cerebral Cortical Hypoperfusion in a Murine Model of Subarachnoid Hemorrhage. Translational Stroke Research. , (2018).
  13. Neulen, A., et al. Neutrophils mediate early cerebral cortical hypoperfusion in a murine model of subarachnoid haemorrhage. Scientific Reports. 9 (1), 8460 (2019).
  14. Neulen, A., et al. Volumetric analysis of intracranial vessels: a novel tool for evaluation of cerebral vasospasm. Int J Comput Assist Radiol Surg. 14 (1), 157-167 (2019).
  15. Washington, C. W., Zipfel, G. J. Participants in the International Multi-disciplinary Consensus Conference on the Critical Care Management of Subarachnoid, H. Detection and monitoring of vasospasm and delayed cerebral ischemia: a review and assessment of the literature. NeuroCritical Care. 15 (2), 312-317 (2011).
  16. Greke, C., et al. Image-guided transcranial Doppler sonography for monitoring of defined segments of intracranial arteries. Journal of Neurosurgical Anesthesiology. 25 (1), 55-61 (2013).
  17. Neulen, A., Prokesch, E., Stein, M., Konig, J., Giese, A. Image-guided transcranial Doppler sonography for monitoring of vasospasm after subarachnoid hemorrhage. Clinical Neurology and Neurosurgery. 145, 14-18 (2016).
  18. Neulen, A., et al. Image-Guided Transcranial Doppler Ultrasound for Monitoring Posthemorrhagic Vasospasms of Infratentorial Arteries: A Feasibility Study. World Neurosurgery. 134, 284-291 (2020).
  19. Neulen, A., et al. Correlation of cardiac function and cerebral perfusion in a murine model of subarachnoid hemorrhage. Scientific Reports. 11 (1), 3317 (2021).
  20. Neulen, A., et al. A segmentation-based volumetric approach to localize and quantify cerebral vasospasm based on tomographic imaging data. PLoS One. 12 (2), 0172010 (2017).
  21. Marbacher, S., et al. Systematic Review of In Vivo Animal Models of Subarachnoid Hemorrhage: Species, Standard Parameters, and Outcomes. Translational Stroke Research. , (2018).
  22. Figueiredo, G., et al. Comparison of digital subtraction angiography, micro-computed tomography angiography and magnetic resonance angiography in the assessment of the cerebrovascular system in live mice. Clinical Neuroradiology. 22 (1), 21-28 (2012).
  23. Lindegaard, K. F., Nornes, H., Bakke, S. J., Sorteberg, W., Nakstad, P. Cerebral vasospasm diagnosis by means of angiography and blood velocity measurements. Acta Neurochirurgica. 100 (1-2), 12-24 (1989).
  24. Cassia, G. S., Faingold, R., Bernard, C., Sant'Anna, G. M. Neonatal hypoxic-ischemic injury: sonography and dynamic color Doppler sonography perfusion of the brain and abdomen with pathologic correlation. American Journal of Roentgenology. 199 (6), 743-752 (2012).
  25. Shen, Q., Stuart, J., Venkatesh, B., Wallace, J., Lipman, J. Inter observer variability of the transcranial Doppler ultrasound technique: impact of lack of practice on the accuracy of measurement. Journal of Clinical Monitoring and Computing. 15 (3-4), 179-184 (1999).

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