A novel atherothrombotic model of ischemic stroke induced by injection of collagen into the cerebral vasculature

J Neurosci Methods. 2015 Jan 15:239:65-74. doi: 10.1016/j.jneumeth.2014.10.001. Epub 2014 Oct 12.

Abstract

Background: Most ischemic strokes in humans are caused by ruptured arterial atheroma, which activate platelets and produce thrombi that occlude cerebral vessels.

Methods: To simulate these events, we threaded a catheter through the internal carotid artery toward the middle cerebral artery (MCA) orifice and injected collagen directly into the cerebral circulation of male C57Bl/6 mice and Wistar rats.

Results: Laser-Doppler flowmetry demonstrated reductions in cerebral blood flow (CBF) of ∼80% in mice and ∼60% in rats. CBF spontaneously increased but remained depressed after catheter withdrawal. Magnetic resonance imaging showed that ipsilateral CBF was reduced at 3h after collagen injection and markedly improved at 48 h. Micro-computed tomography revealed reduced blood vessel density in the ipsilateral MCA territory at 3 h. Gross examination of excised brains revealed thrombi within ipsilateral cerebral arteries at 3 h, but not 24 h, after collagen injection. Immunofluorescence microscopy confirmed that platelets and fibrinogen/fibrin were major components of these thrombi at both macrovascular and microvascular levels. Cerebral infarcts comprising ∼30% of hemispheric volume and neurobehavioral deficits were observed 48 h after ischemic injury in both mice and rats.

Comparison with existing methods: Collagen injection caused brain injury that was similar in magnitude and variability to mechanical MCA occlusion or injection of a pre-formed clot; however, alterations in CBF and the mechanism of vascular occlusion were more consistent with clinical ischemic stroke.

Conclusion: This novel rodent model of ischemic stroke has pathophysiologic characteristics consistent with clinical atherothrombotic stroke, is technically feasible, and creates reproducible brain injury.

Keywords: Atherothrombosis; Collagen; Ischemic stroke; Mouse; Platelet; Rat.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Brain Infarction / chemically induced
  • Brain Ischemia / chemically induced*
  • Brain Ischemia / complications*
  • Cerebrovascular Circulation / drug effects*
  • Cerebrovascular Circulation / physiology
  • Collagen / toxicity*
  • Disease Models, Animal*
  • Functional Laterality
  • Laser-Doppler Flowmetry
  • Magnetic Resonance Imaging
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Nervous System Diseases / etiology
  • Neutrophils / pathology
  • Rats
  • Rats, Wistar
  • Statistics, Nonparametric
  • Stroke / complications
  • Stroke / etiology*
  • Stroke / pathology
  • Time Factors
  • Tomography, X-Ray Computed

Substances

  • Collagen