Abstract
The Wilms’ tumor suppressor gene (WT1) is widely expressed during neovascularization, but it is almost entirely absent in quiescent adult vasculature. However, in vessels undergoing angiogenesis, WT1 is dramatically upregulated. Studies have shown Wt1 has a role in both tumor and ischemic angiogenesis, but the mechanism of Wt1 action in angiogenic tissue remains to be elucidated. Here, we describe two methods for induction of in vivo angiogenesis (subcutaneous sponge implantation, femoral artery ligation) that can be used to assess the influence of Wt1 on new blood vessel formation. Subcutaneously implanted sponges stimulate an inflammatory and fibrotic response including cell infiltration and angiogenesis. Femoral artery ligation creates ischemia in the distal hindlimb and produces an angiogenic response to reperfuse the limb which can be quantified in vivo by laser Doppler flowmetry. In both of these models, the role of Wt1 in the angiogenic process can be assessed using histological/immunohistochemical staining, molecular analysis (qPCR) and flow cytometry. Furthermore, combined with suitable genetic modifications, these models can be used to explore the causal relationship between Wt1 expression and angiogenesis and to trace the lineage of cells expressing Wt1. This approach will help to clarify the importance of Wt1 in regulating neovascularization in the adult, and its potential as a therapeutic target.
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Acknowledgment
R.O. is funded by a British Heart Foundation studentship and RMcG by a Wellcome-Trust-funded ECAT research fellowship. The authors are grateful for support from the Edinburgh British Heart Foundation Centre for Research Excellence (CoRE).
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McGregor, R.J., Ogley, R., Hadoke, P., Hastie, N. (2016). In Vivo Assays for Assessing the Role of the Wilms’ Tumor Suppressor 1 (Wt1) in Angiogenesis. In: Hastie, N. (eds) The Wilms' Tumor (WT1) Gene. Methods in Molecular Biology, vol 1467. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-4023-3_8
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DOI: https://doi.org/10.1007/978-1-4939-4023-3_8
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