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. 2009 Oct 15;183(2):136-40.
doi: 10.1016/j.jneumeth.2009.06.027. Epub 2009 Jul 1.

A novel method for subarachnoid hemorrhage to induce vasospasm in mice

Tamer Altay  1 , Saksith Smithason, Nina Volokh, Peter A Rasmussen, Richard M Ransohoff, J Javier Provencio
Affiliations

A novel method for subarachnoid hemorrhage to induce vasospasm in mice

Tamer Altay et al. J Neurosci Methods. .

Abstract

Mouse models take advantage of genetic manipulations that can be achieved in this species. There are currently two accepted mouse models of subarachnoid hemorrhage (SAH) and cerebral vasospasm (CVs). Both are technically demanding and labor intensive. In this study, we report a reproducible and technically feasible method to induce SAH, and subsequently CVs, in mice. We tested this model in multiple strains of mice that are commonly used for genetic manipulation.

Methods: SAH was induced in C57BL/6NCr, FVB, 129S1, BalbC and SJL mice, weighing 28-32 g, by an intracisternal vessel transection technique. Animals were perfused with India ink at 24h postprocedure and vessel diameters were quantified. Brain slices were obtained for hematoxylin-eosin staining (H&E) to look for vascular changes consistent with CVs.

Results: There was no mortality during or after the procedure. Four of the five mouse strains showed significant CVs at 24 h postprocedure characterized by decreased vessel diameter of the middle cerebral artery close to the Circle of Willis. Histologically, the vessel wall displayed significant corrugation and thickening, consistent with CVs.

Conclusion: A novel mouse model to induce SAH is described and tested in several mouse strains. Four of the five strains used in this study developed CVs after the induction of SAH. The procedure is brief, straightforward, reproducible with low mortality, and applicable to commonly used background strains for genetically engineered mice.

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Figures

Fig. 1
Fig. 1
Procedure for mouse model of SAH. (A) Initial exposure consists of midline suboccipital incision and lateral retraction of strap muscles. (B) A closer view of the underlying transparent atlanto-occipital membrane with the visible subarachnoid vein (arrow). (C) After transection of the vein, blood quickly fills the subarachnoid space. The muscles and skin are closed on one layer and the animals are recovered. (D) After 24 h, there is evidence of blood in the basilar cisterns (arrows). The blood vessels are casted with India ink.
Fig. 2
Fig. 2
Evidence of CVs in an animal with SAH (A) compared to control (D). Note the segmental narrowing of the MCA (closed arrows) and the abrupt interruption of dye in medium vessels (open arrows). Vessels are casted with India ink. H&E preparation of meninges in control (B) shows normal blood vessel wall morphology. In SAH (E) the arterial smooth muscle layer of the MCA artery is thickened and the wall shows rogations consistent to the histology seen in human CVs. When compared to a model of arterial blood injection into the subarachnoid space, the degree of vasospasm by angiography was similar (C) and the arterial thickening was similar although more prominent in the smaller pial vessels than in the MCA (F).
Fig. 3
Fig. 3
Measurement of MCA diameter in five strains of mice (C57B/6J, 129, FVB, BalbC, and SJL). All strains showed decreased mean diameters compared to control suggesting CVs except the SJL strain. Comparisons were made with one-tailed Student's t-test with a p < 0.05 considered significant.
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