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(Stroke. 2003;34:2221.)
© 2003 American Heart Association, Inc.

Original Contributions

Experimental Intracerebral Hemorrhage in the Mouse

Histological, Behavioral, and Hemodynamic Characterization of a Double-Injection Model

Ludmila Belayev, MD; Isabel Saul, BS; Karell Curbelo, MD; Raul Busto, BS; Andrey Belayev, BS; Yongbo Zhang, MD; Panomkhawn Riyamongkol, MS; Weizhao Zhao, PhD Myron D. Ginsberg, MD

Cerebral Vascular Disease Research Center, Department of Neurology, University of Miami School of Medicine, Miami, FL; 33101

Reprint requests to Ludmila Belayev, MD, Department of Neurology (D4-5), University of Miami School of Medicine, PO Box 016960, Miami, FL 33101. E-mail lbelayev{at}stroke.med.miami.edu

Background and Purpose— A major limitation of intracerebral hemorrhage (ICH) research is the lack of reproducible animal models. The present study was conducted to validate in the mouse the double-injection method of ICH initially developed in the rat. We investigated the effect of intrastriatal injection of blood or cerebrospinal fluid (CSF) on cerebral blood flow (CBF), neurological score, hematoma volume, and brain swelling.

Methods— Male C57BL/6 mice were anesthetized with halothane/nitrous oxide delivered by face mask. Rectal and cranial temperatures were regulated at 37°C to 37.5°C. Mice were placed in a stereotactic frame, and a 30-gauge stainless steel cannula was introduced through a burr hole into the left striatum. Each mouse received a 5-µL injection of either whole blood or CSF (over 3 minutes), followed 7 minutes later by 10 µL injected over 5 minutes. The injection cannula was slowly withdrawn 10 minutes after the second injection. Control mice had only cannula insertion. CBF was studied by laser Doppler perfusion imaging. Neurological status was evaluated on days 1 and 2. After 2 days, hematoma volume and brain swelling were calculated.

Results— Physiological values were stable. Mice with ICH but not those with CSF or cannula alone had a marked, persistent neurological deficit and a highly reproducible hematoma, whose mean±SEM volume was 2.0±0.2 mm³ compared with a lesion size of 0.2±0.1 mm³ in mice with CSF. Residual swelling of the ipsilateral hemisphere at 48 hours was 5.7% in the hematoma and 1.5% in the CSF groups. Relative CBF in the neocortex ipsilateral to the injection site declined by 45% to 60% during the first 20 minutes after cannula insertion/injection in all groups but began to renormalize at 25 to 30 minutes in the CSF and cannula-only groups; in the hematoma group, cortical hypoperfusion of 35% to 50% persisted during the 90-minute measurement period.

Conclusions— The present ICH model in mice produces a consistent neurological deficit, hypoperfusion, hematoma volume, and brain swelling. This model closely mimics human hypertensive basal ganglionic ICH and should be useful for the evaluation of pharmaceutical therapies. Laser Doppler perfusion imaging is a useful new technique to quantify relative CBF changes and can be used for studies of dynamic changes of CBF in this in vivo model of ICH in mice.

Key Words: animal models • brain edema • intracerebral hemorrhage • stroke, experimental • mice

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