Micro-computed tomography in murine models of cerebral cavernous malformations as a paradigm for brain disease

J Neurosci Methods. 2016 Sep 15:271:14-24. doi: 10.1016/j.jneumeth.2016.06.021. Epub 2016 Jun 23.

Abstract

Background: Cerebral cavernous malformations (CCMs) are hemorrhagic brain lesions, where murine models allow major mechanistic discoveries, ushering genetic manipulations and preclinical assessment of therapies. Histology for lesion counting and morphometry is essential yet tedious and time consuming. We herein describe the application and validations of X-ray micro-computed tomography (micro-CT), a non-destructive technique allowing three-dimensional CCM lesion count and volumetric measurements, in transgenic murine brains.

New method: We hereby describe a new contrast soaking technique not previously applied to murine models of CCM disease. Volumetric segmentation and image processing paradigm allowed for histologic correlations and quantitative validations not previously reported with the micro-CT technique in brain vascular disease.

Results: Twenty-two hyper-dense areas on micro-CT images, identified as CCM lesions, were matched by histology. The inter-rater reliability analysis showed strong consistency in the CCM lesion identification and staging (K=0.89, p<0.0001) between the two techniques. Micro-CT revealed a 29% greater CCM lesion detection efficiency, and 80% improved time efficiency.

Comparison with existing method: Serial integrated lesional area by histology showed a strong positive correlation with micro-CT estimated volume (r(2)=0.84, p<0.0001).

Conclusions: Micro-CT allows high throughput assessment of lesion count and volume in pre-clinical murine models of CCM. This approach complements histology with improved accuracy and efficiency, and can be applied for lesion burden assessment in other brain diseases.

Keywords: Cerebral cavernous malformation; Iodine contrast; Micro-CT; Morphometry.

Publication types

  • Validation Study

MeSH terms

  • Animals
  • Apoptosis Regulatory Proteins
  • Brain / diagnostic imaging*
  • Contrast Media
  • Disease Models, Animal*
  • Female
  • Hemangioma, Cavernous, Central Nervous System / diagnostic imaging*
  • Histological Techniques
  • Intracellular Signaling Peptides and Proteins / genetics
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Iodine
  • Male
  • Mice
  • Mice, Transgenic
  • Observer Variation
  • Organ Size
  • Reproducibility of Results
  • Time Factors
  • X-Ray Microtomography / methods*
  • rho-Associated Kinases / genetics
  • rho-Associated Kinases / metabolism

Substances

  • Apoptosis Regulatory Proteins
  • Contrast Media
  • Intracellular Signaling Peptides and Proteins
  • PDCD10 protein, mouse
  • Iodine
  • Rock1 protein, mouse
  • Rock2 protein, mouse
  • rho-Associated Kinases