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Neuroscience

Modeling Stroke in Mice: Transient Middle Cerebral Artery Occlusion via the External Carotid Artery

Published: May 24th, 2021

DOI:

10.3791/62573

1Institute for Stroke and Dementia Research, LMU Munich, 2Munich Cluster for Systems Neurology (SyNergy)

Different models of middle cerebral artery occlusion (MCAo) are used in experimental stroke research. Here, an experimental stroke model of transient MCAo via the external carotid artery (ECA) is described, which aims to mimic human stroke, in which the cerebrovascular thrombus is removed due to spontaneous clot lysis or therapy.

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options are limited to a small proportion of stroke patients within the first hours after stroke. Novel therapeutic strategies are being extensively investigated, especially to prolong the therapeutic time window. These current investigations include the study of important pathophysiological pathways after stroke, such as post-stroke inflammation, angiogenesis, neuronal plasticity, and regeneration. Over the last decade, there has been increasing concern about the poor reproducibility of experimental results and scientific findings among independent research groups. To overcome the so-called “replication crisis”, detailed standardized models for all procedures are urgently needed. As an effort within the “ImmunoStroke” research consortium (https://immunostroke.de/), a standardized mouse model of transient middle cerebral artery occlusion (MCAo) is proposed. This model allows the complete restoration of the blood flow upon removal of the filament, simulating the therapeutic or spontaneous clot lysis that occurs in a large proportion of human strokes. The surgical procedure of this “filament” stroke model and tools for its functional analysis are demonstrated in the accompanying video.

Stroke is one of the most common causes of death and disability worldwide. Although there are mainly two distinct forms of stroke, ischemic and hemorrhagic, 80–85% of all stroke cases are ischemic1. Currently, only two treatments are available for patients with ischemic stroke: pharmacological treatment with recombinant tissue plasminogen activator (rtPA) or mechanical thrombectomy. However, due to the narrow therapeutic time window and multiple exclusion criteria, only a select number of patients can benefit from these specific treatment options. Over the last two decades, preclinical and translational stroke research has focused on the ....

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The experiments reported in this video were conducted following the national guidelines for the use of experimental animals, and the protocols were approved by the German governmental committees (Regierung von Oberbayern, Munich, Germany). Ten-week-old male C57Bl/6J mice were used and housed under controlled temperature (22 ± 2 °C), with a 12 h light-dark cycle period and access to pelleted food and water ad libitum.

1. Preparation of the material and instruments

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The model described here is a modification of the commonly used "filament" stroke model, which consists of introducing a silicon-coated filament through the ECA to transiently block the origin of the MCA (Figure 1). After removing the filament, only the blood flow in the ECA is permanently ceased, allowing complete recanalization of the CCA and ICA. This allows an adequate reperfusion of the brain (Figure 2), similar to the situation observed after succe.......

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The present protocol describes an experimental stroke model based on the consensus agreement of a German multicenter research consortium (“ImmunoStroke”) to establish a standardized transient MCAo model. The transient MCAo model established by introducing a silicon-coated filament through the ECA to the origin of the MCA is one of the most widely used stroke models to achieve arterial reperfusion after a delimitated occlusion period. Therefore, this procedure can be considered a translationally relevant strok.......

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We thank all our collaboration partners of the ImmunoStroke Consortia (FOR 2879, From immune cells to stroke recovery) for suggestions and discussions. This work was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany's Excellence Strategy within the framework of the Munich Cluster for Systems Neurology (EXC 2145 SyNergy - ID 390857198) and under the grants LI-2534/6-1, LI-2534/7-1 and LL-112/1-1.

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Name Company Catalog Number Comments
45° ramp H&S Kunststofftechnik height: 18 cm
5/0 threat Pearsalls 10C103000
5 mL Syringe Braun
Acetic Acid Sigma Life Science 695092
Anesthesia system for isoflurane Drager
Bepanthen pomade Bayer
C57Bl/6J mice Charles River 000664
Clamp FST 12500-12
Clip FST 18055-04
Clip holder FST 18057-14
Cotons NOBA Verbondmitel Danz 974116
Cresyl violet Sigma Life Science C5042-10G
Cryostat Thermo Scientific CryoStarNX70
Ethanol 70% CLN Chemikalien Laborbedorf 521005
Ethanol 96% CLN Chemikalien Laborbedorf 522078
Ethanol 99% CLN Chemikalien Laborbedorf ETO-5000-99-1
Filaments Doccol 602112PK5Re
Fine 45 angled forceps FST 11251-35
Fine forceps FST 11252-23
Fine Scissors FST 14094-11
Glue Orechseln BSI-112
Hardener Glue Drechseln & Mehr BSI-151
Heating blanket FHC DC Temperature Controller
Isoflurane Abbot B506
Isopentane Fluka 59070
Ketamine Inresa Arzneimittel GmbH
Laser Doppler Perimed PF 5010 LDPM, Periflux System 5000
Laser Doppler probe Perimed 91-00123
Phosphate Buffered Saline pH: 7.4 Apotheke Innestadt Uni Munchen P32799
Recovery chamber Mediheat
Roti-Histokit mounting medium Roth 6638.1
Saline solution Braun 131321
Scalpel Feather 02.001.30.011
Silicon-coated filaments Doccol 602112PK5Re
Stereomicropscope Leica M80
Superfrost Plus Slides Thermo Scientific J1800AMNZ
Vannas Spring Scissors FST 15000-00
Xylacine Albrecht

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