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Adipose angiogenesis: quantitative methods to study microvessel growth, regression and remodeling in vivo

Nature Protocols volume 5, pages 912–920 (2010)Cite this article

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Abstract

Genetic and diet-induced rodent obesity models provide outstanding opportunities to study the role of angiogenesis and vascular remodeling in modulation of adipogenesis and obesity. In this study, we describe methods to quantitatively study adipose angiogenesis and vascular remodeling on the basis of immunohistochemical analyses. Fresh white adipose tissue or brown adipose tissue are prepared for whole mount, cryosectioned and paraffin-embedded samples, followed by staining with specific markers such as platelet endothelial cell adhesion molecule-1 (PECAM-1)/CD31, CD34, isolectin B4 or α-smooth muscle actin. Adipocytes are visualized by staining lipid droplets with 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene-3-dodecanoic acid (BODIPY) 558/568 C12. This protocol may take 2–5 d to obtain results. In the view of the crucial roles of vasculature in modulation of adipogenesis and obesity, this protocol is valuable for studying the molecular mechanisms of angiogenesis in obese adipose tissues and for assessing the anti-obesity activity of angiogenesis modulators.

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Figure 1: Comparison of various blood and lymphatic vascular endothelial cell markers in WAT.
Figure 2: Comparative evaluation of vascular structure and density in WAT and BAT.
Figure 3: Analysis of vascular remodeling and functions in adipose tissue.

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Acknowledgements

Yihai Cao's laboratory is supported through research grants from the Swedish Research Council, the Swedish Cancer Foundation, the Karolinska Institute Foundation, the Karolinska Institute Distinguished Professor Award, the European Union Integrated Project of Metoxia (Project no. 222741) and the European Research Council (ERC) advanced grant ANGIOFAT (Project no. 250021).

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  1. Yuan Xue and Sharon Lim: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden

    Yuan Xue, Sharon Lim & Yihai Cao

  2. Institute for Biomedical Research, Rouen Medical University, Rouen, France

    Ebba Bråkenhielm

Authors
  1. Yuan Xue
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  2. Sharon Lim
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  3. Ebba Bråkenhielm
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Contributions

Y.C. designed the study. Y.X. and S.L. performed and analyzed experiments. E.B. contributed to the development of these protocols. Y.X., S.L. and Y.C. wrote the paper.

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Correspondence to Yihai Cao.

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The authors declare no competing financial interests.

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Xue, Y., Lim, S., Bråkenhielm, E. et al. Adipose angiogenesis: quantitative methods to study microvessel growth, regression and remodeling in vivo. Nat Protoc 5, 912–920 (2010). https://doi.org/10.1038/nprot.2010.46

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