The CCB was amongst the first to generate knockout mice in the early 1990s and has, since then generated dozens of transgenic (disease) mouse models. Since more than 15 years, the CCB has invested substantial resources to develop an "in house" mouse clinic to phenotype these mouse models extensively, with the aspiration to use similar (but adapted) tools, assays and techniques as those commonly used in the clinic to diagnose diseases in humans. Therefore, assays and techniques, initially only available for larger animal species, were miniaturized and adapted, and "mouse doctors" with microsurgical skills and expertise were trained. Over the years, the CCB has been pioneering the development and use of novel assays and models in neurovascular research. An overview of such assays is listed.
Angiogenesis & lymphangiogenesis
Angiogenesis is critical for normal development but also contributes to the pathogenesis of numerous diseases.
We routinely study vascular development in the embryo and placenta (whole mount immunostaining & in situ hybridization) and the postnatal retina and other organs (microvascular casting), and analyze the remodeling of the large thoracic vessels (intracardial angiography), the maintenance or regression of airway vessels.
Angiogenesis in pathological conditions is studied in available mouse models of cancer (ectopic or orthotopic implantation of syngeneic or xenograft tumors, spontaneously developing tumor models, hematogenic and lymphatic metastasis models), myocardial infarction (ligation of coronary artery and vene), post-infarct myocardial hypertrophy, hindlimb ischemia (ligation of femoral artery), eye disease (choroid neovascularization, retinal oxygen-induced vascular proliferation, cornea pocket neovascularization, neovascularization after corneal cauterization), skin wound healing (healthy and diabetic mice), matrigel plug neovascularization, inflammatory disease (models of colitis), liver disease (cirrhosis) and others.
Assays are available to study various aspects of the structure and functionality of vessels, i.e. vascular permeability and leakage, perfusion (microspheres), drug delivery, (delivery of inhibitors via tail vein, retro-orbital, intratracheal, intravitreal or intra-amniotic injection), etc.
The following assays are available to study metabolism (in house or via local collaborations): measurements of metabolites in plasma and tissues (lactate, glycogen, FFA, ketone bodies, etc); 13C-NMR spectroscopy to measure glucose and fatty acid oxidation ex vivo; 31P-NMR spectroscopy to measure high energy phosphates (ATP, etc) in vivo; 1H-NMR spectroscopy to measure lactate and succinate ex vivo; EPR oxymetry to measure oxygen consumption; tracer studies to measure glycolysis, glucose and fatty acid oxidation in muscle preparations ex vivo; micro-PET measurements of glucose uptake; measurement of mitochondrial respiration in isolated mitochondria; etc. Isolation setups for murine endothelial cell in different organs are in place, allowing the study of the in metabolism of murine endothelial cells in vitro.
A number of other mouse models are available: ischemia/reperfusion (in kidney, heart, liver and hindlimb); pulmonary hypertension (hypoxia chambers; thrombosis and thrombolysis (pulmonary embolization via collagen-epinephrin injection, endothelial damage via Rose-Bengal injection,; pulmonary thrombolysis model), electroporation in utero.
- Wong BW et al. The role of fatty acid ﬂ-oxidation in lymphangiogenesis. Nature. 2017 Feb 2;542(7639):49-54
- Cantelmo AR et al. Inhibition of the Glycolytic Activator PFKFB3 in Endothelium Induces Tumor Vessel Normalization, Impairs Metastasis, and Improves Chemotherapy. Cancer Cell. 2016 Dec 12;30(6):968-985.
- Schoors S et al. Fatty acid carbon is essential for dNTP synthesis in endothelial cells. Nature. 2015 Apr 9;520(7546):192-197.