Name: analysis of cerebral vasospasm in a murine model of subarachnoid hemorrhage with high frequency transcranial duplex ultrasound
Uploaded: 2021-06-03T14:00:00
Description: Watch this Scientific Journal Video about Analysis of Cerebral Vasospasm in a Murine Model of Subarachnoid Hemorrhage with High Frequency Transcranial Duplex Ultrasound at JoVE.com

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 Kr&#246;ner-Fresenius-Stiftung (2020_EKEA.144).

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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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...

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 caused by microvasospasms and microthrombosis, cortical spreading depressions, and effects triggered by inflammation4. Unfortunately, the exact pathophysiology remains unclear and there is no treatment available that effectively prevents DCI3. Therefore, DCI is investigated in many clinical and experimental studies.
Nowadays, most experimental studies on SAH use small animal models, especially in mice5,6,7,8,9,10,11,12,13. In such studies, cerebral vasospasm is frequently investigated as an endpoint. It is common to determine the degree of vasospasm ex vivo. This is because noninvasive methods for in vivo examination of cerebral vasospasm requiring short anesthesia time and imposing only little distress on the animals are lacking. However, examination of cerebral vasospasm in vivo would be advantageous. This is because it would allow longitudinal in vivo studies on vasospasm in mice (i.e., imaging of cerebral vasospasm at different time points during the days after induction of SAH). This would enhance the comparability of data acquired at different time points. Furthermore, using a longitudinal study design is a strategy to reduce animal numbers.
Here we demonstrate the use of high frequency transcranial ultrasound to determine the blood flow in cerebral arteries in mice. We show that, similar to transcranial Doppler sonography (TCD) or transcranial color-coded Duplex sonography (TCCD) in clinical practice14,15,16,17,18, this method can be used to monitor cerebral vasospasm by measuring the blood flow velocities of the intracranial arteries after SAH induction in the murine model.

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			<td>C57BL/6N mice</td>
			<td>Janvier; Saint-Berthevin Cedex, France</td>
			<td>n.a.</td>
			<td>mice</td>
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			<td>Groppler; Oberperasberg, Germany</td>
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			<td>isoflurane vaporizer</td>
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			<td>VisualSonics; FujiFilm, Toronto, CA</td>
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analysis of cerebral vasospasm in a murine model of subarachnoid hemorrhage with high frequency transcranial duplex ultrasound

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.

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 &#176;C19.
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Watch this Scientific Journal Video about Analysis of Cerebral Vasospasm in a Murine Model of Subarachnoid Hemorrhage with High Frequency Transcranial Duplex Ultrasound at JoVE.com

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

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 ...

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