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A protocol for a lung neovascularization model in rodents

Nature Protocols volume 3, pages 378–387 (2008)Cite this article

Abstract

By providing insight into the cellular events of vascular injury and repair, experimental model systems seek to promote timely therapeutic strategies for human disease. The goal of many current studies of neovascularization is to identify cells critical to the process and their role in vascular channel assembly. We propose here a protocol to analyze, in an in vivo rodent model, vessel and capillary remodeling (reorganization and growth) in the injured lung. Sequential analyses of stages in the assembly of vascular structures, and of relevant cell types, provide further opportunities to study the molecular and cellular determinants of lung neovascularization.

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Figure 1: Equipment setup.
Figure 2: Drawing illustrating the inflation apparatus to distend lungs initially with a 3% paraformaldehyde/0.1% gluteraldehyde solution (shown in blue).
Figure 3: Image and cartoon of fixative-distended rodent (rat) lung.
Figure 4: Illustration of vessel wall thickening by brightfield microscopy and by TEM in normal rat lung and in the rat lung after breathing high oxygen.
Figure 5: Illustration of alveolar-capillary membrane structure in the normal rat lung and in the rat lung after breathing high oxygen.

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References

  1. Jones, R.C. & Jacobson, M. Angiogenesis in the hypertensive lung: response to ambient oxygen tension. Cell Tissue Res. 300, 263–284 (2000).

    Article  CAS  Google Scholar 

  2. Jones, R., Steudel, W., White, S., Jacobson, M. & Low, R. Microvessel precursor smooth muscle cells express head-inserted smooth muscle myosin heavy chain (SM-B) isoform in hyperoxic pulmonary hypertension. Cell Tissue Res. 295, 453–465 (1999).

    Article  CAS  Google Scholar 

  3. Jones, R., Jacobson, M. & Steudel, W. alpha-smooth-muscle actin and microvascular precursor smooth-muscle cells in pulmonary hypertension. Am. J. Respir. Cell Mol. Biol. 20, 582–594 (1999).

    Article  CAS  Google Scholar 

  4. Jones, R. Ultrastructural analysis of contractile cell development in lung microvessels in hyperoxic pulmonary hypertension. Fibroblasts and intermediate cells selectively reorganize nonmuscular segments. Am. J. Pathol. 141, 1491–1505 (1992).

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Jones, R.C. et al. A protocol for phenotypic detection and characterization of vascular cells of different origins in a lung neovascularization model in rodents. Nat. Protoc. 3, 388–397 (2008).

    Article  CAS  Google Scholar 

  6. Mitzner, W. & Wagner, E.M. Vascular remodeling in the circulations of the lung. J. Appl. Physiol. 97, 1999–2004 (2004).

    Article  Google Scholar 

  7. Jones, R. & Reid, L. Vascular remodeling in clinical and experimental pulmonary hypertensions. in Pulmonary Vascular Remodeling 47–115 (Portland Press, London, 1995).

    Google Scholar 

  8. Frid, M.G. et al. Hypoxia-induced pulmonary vascular remodeling requires recruitment of circulating mesenchymal precursors of a monocyte/macrophage lineage. Am. J. Pathol. 168, 659–669 (2006).

    Article  CAS  Google Scholar 

  9. Boxt, L.M. et al. Fractal analysis of pulmonary arteries: the fractal dimension is lower in pulmonary hypertension. J. Thorac. Imaging 9, 8–13 (1994).

    Article  CAS  Google Scholar 

  10. Hu, L.M. & Jones, R. Injury and remodeling of pulmonary veins by high oxygen. A morphometric study. Am. J. Pathol. 134, 253–262 (1989).

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Klekamp, J.G., Jarzecka, K. & Perkett, E.A. Exposure to hyperoxia decreases the expression of vascular endothelial growth factor and its receptors in adult rat lungs. Am. J. Pathol. 154, 823–831 (1999).

    Article  CAS  Google Scholar 

  12. Maniscalco, W.M., Watkins, R.H., Finkelstein, J.N. & Campbell, M.H. Vascular endothelial growth factor mRNA increases in alveolar epithelial cells during recovery from oxygen injury. Am. J. Respir. Cell Mol. Biol. 13, 377–386 (1995).

    Article  CAS  Google Scholar 

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Acknowledgements

Supported by NIH R01HL 070866 (to R.C.J.), NIH R01CA 115767 (to R.K.J.) and by a Career Development Award to D.G.D. (from the American Association for Cancer Research-Genentech BioOncology).

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Authors and Affiliations

  1. Department of Anesthesia and Critical Care, Massachusetts General Hospital and Harvard Medical School, 55 Fruit Street, Boston, 02114, Massachusetts, USA

    Rosemary C Jones, Diane Capen & Bodil Petersen

  2. Steele Laboratory for Tumor Biology, Massachusetts General Hospital and Harvard Medical School, 55 Fruit Street, Boston, 02114, Massachusetts, USA

    Rakesh K Jain & Dan G Duda

Authors
  1. Rosemary C Jones
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  2. Diane Capen
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  3. Bodil Petersen
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  4. Rakesh K Jain
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  5. Dan G Duda
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Correspondence to Rosemary C Jones.

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Jones, R., Capen, D., Petersen, B. et al. A protocol for a lung neovascularization model in rodents. Nat Protoc 3, 378–387 (2008). https://doi.org/10.1038/nprot.2007.536

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