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Summary

Abstract

Introduction

Protocol

Representative Results

Discussion

Acknowledgements

Materials

References

Neuroscience

Protocol for Isolating the Mouse Circle of Willis

Published: October 22nd, 2016

DOI:

10.3791/54352

1UPMC Univ Paris 06, Sorbonne Universités

We describe here a reproducible protocol for isolating the mouse circle of Willis.

The cerebral arterial circle (circulus arteriosus cerebri) or circle of Willis (CoW) is a circulatory anastomosis surrounding the optic chiasma and hypothalamus that supplies blood to the brain and surrounding structures. It has been implicated in several cerebrovascular disorders, including cerebral amyloid angiopathy (CAA)-associated vasculopathies, intracranial atherosclerosis and intracranial aneurysms. Studies of the molecular mechanisms underlying these diseases for the identification of novel drug targets for their prevention require animal models. Some of these models may be transgenic, whereas others will involve isolation of the cerebro-vasculature, including the CoW.The method described here is suitable for CoW isolation in any mouse lineage and has considerable potential for screening (expression of genes, protein production, posttranslational protein modifications, secretome analysis, etc.) studies on the large vessels of the mouse cerebro-vasculature. It can also be used for ex vivo studies, by adapting the organ bath system developed for isolated mouse olfactory arteries.

The cerebral arterial circle (circulus arteriosus cerebri), also known as the circle of Willis (CoW), loop of Willisor Willis polygon) was first described by Thomas Willis in 1664. It is a circulatory anastomosis located around the optic chiasma and hypothalamus that can be considered as a central hub supplying blood to the brain and surrounding structures. Blood enters this structure via the internal carotid and vertebral arteries and it flows out of the circle via the interior middle and posterior cerebral arteries. Each of these arteries has left and right branches on either side of the circle. The basilar, post communicating, and anterior communicating arteries co....

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All procedures were performed in accordance with European Community standards for the care and use of laboratory animals, with the approval of the local ethics committee for animal experimentation (Ile de France-Paris-Committee, Authorization 4270).

1. Anesthesia

  1. Infuse a lethal dose of pentobarbital (up to 1 mg/10 g body weight) intraperitoneally (27-gauge needle and 1-ml syringe) into adult mice before surgery.

2. Vessel Perfusion

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The PBS-perfused mouse is killed and the CoW is isolated as described in section 3.2 of the protocol. When the dissection is performed correctly, the CoW should come out in one piece and should be slightly transparent due to the absence of residual blood in the vasculature.

Figure 2
Figure 2: The Mouse CoW after Isolation. (A) Overv.......

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We describe here a reproducible protocol for the isolation of the circle of Willis. The most common cerebrovascular disorders involving the CoW are CAA-associated vasculopathies, intracranial atherosclerosis and intracranial aneurysm, all of which affect the walls of arterial vessels. The risk factors are well known, but the molecular pathogenesis of these cerebral disorders remains poorly understood and specific biological markers for predicting their occurrence are lacking. There is considerable interest in methods for.......

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This work was supported by Paris VI University and a Pierre Fabre Innovation grant.

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Name Company Catalog Number Comments
Dulbecco’s Phosphate Buffered Saline Sigma-Aldrich D8537
Dumont #55 Forceps Fine Science Tools 11295-51
Hardened Fine Iris Scissors  Fine Science Tools 14090-11
Scissors - Straight / Sharp / Sharp   16.5 cm Fine Science Tools 14002-16
Dumont #7b Forceps  Fine Science Tools 11270-20
Stereoscopic Zoom Microscope Nikon SMZ745T
CellBIND Surface 60mm Culture Dish Corning #3295
Peristaltic Pump - MINIPULS 3 Gilson M312
Pentobarbital Sodique Ceva Santé Animale FR/V/2770465 3/1992

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