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Angiogenesis in Adipose Tissue

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Adipose Tissue Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 456))

Summary

Angiogenesis is required for the growth and expansion of both healthy and pathological tissues. The plasticity of the adipose tissue is reflected by its remarkable ability to expand or to reduce in size throughout the adult lifespan. We, and others, have recently shown that expansion of fat mass is dependent on angiogenesis, and suppression of angiogenesis might provide a novel therapeutic approach for prevention and treatment of obesity. Here, we outline two technical procedures for assessment of angiogenesis in adipose tissues.

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References

  1. Di Girolamo M, Skinner NS Jr., Hanley HG, Sachs RG (1971) Relationship of adipose tissue blood flow to fat cell size and number. Am J Physiol 220:932–937

    CAS  PubMed  Google Scholar 

  2. Crandall DL, Hausman GJ, Kral JG (1997) A review of the microcirculation of adipose tissue: anatomic, metabolic, and angiogenic perspectives. Microcirculation 4:211–232

    Article  CAS  PubMed  Google Scholar 

  3. Gersh I, Still MA (1945) Blood vessels in fat tissue: relation to problems of gas exchange. J Exp Med 81:219–232

    Article  CAS  PubMed  Google Scholar 

  4. Cinti S (2005) The adipose organ. Prostaglandins Leukot Essent Fatty Acids 73:9–15

    Article  CAS  PubMed  Google Scholar 

  5. Risau W (1997) Mechanisms of angiogenesis. Nature 386:671–674

    Article  CAS  PubMed  Google Scholar 

  6. Carmeliet P (2000) Mechanisms of angiogenesis and arteriogenesis. Nat Med 6:389–395

    Article  CAS  PubMed  Google Scholar 

  7. Hausman GJ, Richardson RL (1983) Cellular and vascular development in immature rat adipose tissue. J Lipid Res 24:522–532

    CAS  PubMed  Google Scholar 

  8. Poissonnet CM, LaVelle M, Burdi AR (1988) Growth and development of adipose tissue. J Pediatr 113:1–9

    Article  CAS  PubMed  Google Scholar 

  9. Kimura Y, Ozeki M, Inamoto T, Tabata Y (2002) Time course of de novo adipogenesis in matrigel by gelatin microspheres incorporating basic fibroblast growth factor. Tissue Eng 8:603–613

    Article  CAS  PubMed  Google Scholar 

  10. Sypniewska G, Bjorntorp P (1987) Increased DNA synthesis in adipocytes and capillary endothelium in rat adipose tissue during overfeeding. Eur J Clin Invest 17:202–207

    Article  CAS  PubMed  Google Scholar 

  11. Krotkiewski M, Bjorntorp P, Sjostrom L, Smith U (1983) Impact of obesity on metabolism in men and women. Importance of regional adipose tissue distribution. J Clin Invest 72:1150–1162

    Article  CAS  PubMed  Google Scholar 

  12. Di Girolamo M, Mendlinger S, Fertig JW (1971) A simple method to determine fat cell size and number in four mammalian species. Am J Physiol 221:850–858

    CAS  PubMed  Google Scholar 

  13. Rupnick MA, Panigrahy D, Zhang C Y, Dallabrida SM, Lowell BB, Langer R, Folkman MJ (2002) Adipose tissue mass can be regulated through the vasculature. Proc Natl Acad Sci U S A 99:10730–10735

    Article  CAS  PubMed  Google Scholar 

  14. Brakenhielm E, Cao R, Gao B, Angelin B, Cannon B, Parini P, Cao Y (2004) Angiogenesis inhibitor, TNP-470, prevents diet-induced and genetic obesity in mice. Circ Res 94:1579–1588

    Article  CAS  PubMed  Google Scholar 

  15. Dallabrida SM, Zurakowski D, Shih SC, Smith LE, Folkman J, Moulton KS, Rupnick MA (2003) Adipose tissue growth and regression are regulated by angiopoietin-1. Biochem Biophys Res Commun 311:563–571

    Article  CAS  PubMed  Google Scholar 

  16. Liu L, Meydani M (2003) Angiogenesis inhibitors may regulate adiposity. Nutr Rev 61: 384–387

    Article  PubMed  Google Scholar 

  17. Yamaguchi M, Matsumoto F, Bujo H, Shibasaki M, Takahashi K, Yoshimoto S, Ichinose M, Saito Y (2005) Revascularization determines volume retention and gene expression by fat grafts in mice. Exp Biol Med (Maywood) 230:742–748

    CAS  Google Scholar 

  18. Fukumura D, Ushiyama A, Duda DG, Xu L, Tam J, Krishna V, Chatterjee K, Garkavtsev I, Jain RK (2003) Paracrine regulation of angiogenesis and adipocyte differentiation during in vivo adipogenesis. Circ Res 93: e88–e97

    Article  CAS  PubMed  Google Scholar 

  19. Tabata Y, Miyao M, Inamoto T, Ishii T, Hirano Y, Yamaoki Y, Ikada Y (2000) De novo formation of adipose tissue by controlled release of basic fibroblast growth factor. Tissue Eng 6:279–289

    Article  CAS  PubMed  Google Scholar 

  20. Vashi AV, Abberton KM, Thomas GP, Morrison WA, O'Connor A, J., Cooper-White JJ, Thompson EW (2006) Adipose tissue engineering based on the controlled release of fibroblast growth factor-2 in a collagen matrix. Tissue Eng 12:3035–3043

    Article  CAS  PubMed  Google Scholar 

  21. Kawaguchi N, Toriyama K, Nicodemou-Lena E, Inou K, Torii S, Kitagawa Y (1998) De novo adipogenesis in mice at the site of injection of basement membrane and basic fibroblast growth factor. Proc Natl Acad Sci U S A 95:1062–1066

    Article  CAS  PubMed  Google Scholar 

  22. Castellot JJ Jr., Karnovsky MJ, Spiegelman BM (1982) Differentiation-dependent stimulation of neovascularization and endothelial cell chemotaxis by 3T3 adipocytes. Proc Natl Acad Sci U S A 79:5597–5601

    Article  PubMed  Google Scholar 

  23. Castellot JJ Jr., Karnovsky MJ, Spiegelman BM (1980) Potent stimulation of vascular endothelial cell growth by differentiated 3T3 adipocytes. Proc Natl Acad Sci U S A 77:6007–6011

    Article  CAS  PubMed  Google Scholar 

  24. Folkman J (1982) Angiogenesis: initiation and control. Ann N Y Acad Sci 401:212–227

    Article  CAS  PubMed  Google Scholar 

  25. Goldsmith HS, Griffith AL, Kupferman A, Catsimpoolas N (1984) Lipid angiogenic factor from omentum. JAMA 252:2034–2036

    Article  CAS  PubMed  Google Scholar 

  26. Silverman KJ, Lund DP, Zetter BR, Lainey LL, Shahood JA, Freiman DG, Folkman J, Barger AC (1988) Angiogenic activity of adipose tissue. Biochem Biophys Res Commun 153:347–352

    Article  CAS  PubMed  Google Scholar 

  27. Zhang QX, Magovern CJ, Mack CA, Budenbender KT, Ko W, Rosengart TK (1997) Vascular endothelial growth factor is the major angiogenic factor in omentum: mechanism of the omentum-mediated angiogenesis. J Surg Res 67:147–154

    Article  CAS  PubMed  Google Scholar 

  28. Rehman J, Considine RV, Bovenkerk JE, Li J, Slavens CA, Jones RM, March KL (2003) Obesity is associated with increased levels of circulating hepatocyte growth factor. J Am Coll Cardiol 41:1408–1413

    Article  CAS  PubMed  Google Scholar 

  29. Saiki A, Watanabe F, Murano T, Miyashita Y, Shirai K (2006) Hepatocyte growth factor secreted by cultured adipocytes promotes tube formation of vascular endothelial cells in vitro. Int J Obes (Lond) 30:1676–1684

    Article  CAS  Google Scholar 

  30. Dobson DE, Kambe A, Block E, Dion T, Lu H, Castellot JJ Jr., Spiegelman BM (1990) 1-Butyryl-glycerol: a novel angiogenesis factor secreted by differentiating adipocytes. Cell 61:223–230

    Article  CAS  PubMed  Google Scholar 

  31. Asano A, Kimura K, Saito M (1999) Cold-induced mRNA expression of angiogenic factors in rat brown adipose tissue. J Vet Med Sci 61:403–409

    Article  CAS  PubMed  Google Scholar 

  32. Asano A, Irie Y, Saito M (2001) Isoform-specific regulation of vascular endothelial growth factor (VEGF) family mRNA expression in cultured mouse brown adipocytes. Mol Cell Endocrinol 174:71–76

    Article  CAS  PubMed  Google Scholar 

  33. Silha J V, Krsek M, Sucharda P, Murphy LJ (2005) Angiogenic factors are elevated in overweight and obese individuals. Int J Obes (Lond) 29:1308–1314

    Article  CAS  Google Scholar 

  34. Stacker SA, Runting AS, Caesar C, Vitali A, Lackmann M, Chang J, Ward L, Wilks AF (2000) The 3T3-L1 fibroblast to adipocyte conversion is accompanied by increased expression of angiopoietin-1, a ligand for tie2. Growth Factors 18:177–191

    Article  CAS  PubMed  Google Scholar 

  35. Voros G, Maquoi E, Demeulemeester D, Clerx N, Collen D, Lijnen HR (2005) Modulation of angiogenesis during adipose tissue development in murine models of obesity. Endocrinology 146:4545–4554

    Article  CAS  PubMed  Google Scholar 

  36. Teichert-Kuliszewska K, Hamilton BS, Deitel M, Roncari DA (1992) Augmented production of heparin-binding mitogenic proteins by preadipocytes from massively obese persons. J Clin Invest 90:1226–1231

    Article  CAS  PubMed  Google Scholar 

  37. Lijnen HR, Christiaens V, Scroyen I, Voros G, Tjwa M, Carmeliet P, Collen D (2006) Impaired adipose tissue development in mice with inactivation of placental growth factor function. Diabetes 55:2698–2704

    Article  CAS  PubMed  Google Scholar 

  38. Cao R, Brakenhielm E, Wahlestedt C, Thyberg J, Cao Y (2001) Leptin induces vascular permeability and synergistically stimulates angiogenesis with FGF-2 and VEGF. Proc Natl Acad Sci U S A 98:6390–6395

    Article  CAS  PubMed  Google Scholar 

  39. Bouloumie A, Drexler HC, Lafontan M, Busse R (1998) Leptin, the product of Ob gene, promotes angiogenesis. Circ Res 83:1059–1066

    CAS  PubMed  Google Scholar 

  40. Cleland WH, Mendelson CR, Simpson ER (1983) Aromatase activity of membrane fractions of human adipose tissue stromal cells and adipocytes. Endocrinology 113:2155–2160

    Article  CAS  PubMed  Google Scholar 

  41. Ichinose Y, Asoh H, Yano T, Yokoyama H, Inoue T, Tayama K, Ueda T, Takai E (1995) Use of a pericardial fat pad flap for preventing bronchopleural fistula: an experimental study focusing on the angiogenesis and cytokine production of the fat pad. Surg Today 25: 811–815

    Article  CAS  PubMed  Google Scholar 

  42. Guan H, Arany E, van Beek JP, Chamson-Reig A, Thyssen S, Hill DJ, Yang K (2005) Adipose tissue gene expression profiling reveals distinct molecular pathways that define visceral adiposity in offspring of maternal protein-restricted rats. Am J Physiol Endocrinol Metab 288: E663–E673

    Article  CAS  PubMed  Google Scholar 

  43. Rehman J, Traktuev D, Li J, Merfeld-Clauss S, Temm-Grove CJ, Bovenkerk JE, Pell CL, Johnstone BH, Considine RV, March KL (2004) Secretion of angiogenic and antiapoptotic factors by human adipose stromal cells. Circulation 109:1292–1298

    Article  PubMed  Google Scholar 

  44. Samad F, Pandey M, Loskutoff DJ (1998) Tissue factor gene expression in the adipose tissues of obese mice. Proc Natl Acad Sci U S A 95:7591–7596

    Article  CAS  PubMed  Google Scholar 

  45. Bouloumie A, Sengenes C, Portolan G, Galitzky J, Lafontan M (2001) Adipocyte produces matrix metalloproteinases 2 and 9: involvement in adipose differentiation. Diabetes 50:2080–2086

    Article  CAS  PubMed  Google Scholar 

  46. Tartare-Deckert S, Chavey C, Monthouel MN, Gautier N, Van Obberghen E (2001) The matri-cellular protein SPARC/osteonectin as a newly identified factor up-regulated in obesity. J Biol Chem 276:22231–22237

    Article  CAS  PubMed  Google Scholar 

  47. Li J, Yu X, Pan W, Unger RH (2002) Gene expression profile of rat adipose tissue at the onset of high-fat-diet obesity. Am J Physiol Endocrinol Metab 282: E1334–E1341

    CAS  PubMed  Google Scholar 

  48. Taleb S, Lacasa D, Bastard JP, Poitou C, Cancello R, Pelloux V, Viguerie N, Benis A, Zucker JD, Bouillot JL, Coussieu C, Basdevant A, Langin D, Clement K (2005) Cathepsin S, a novel biomarker of adiposity: relevance to atherogenesis. FASEB J 19:1540–1542

    CAS  PubMed  Google Scholar 

  49. Brakenhielm E, Veitonmaki N, Cao R, Kihara S, Matsuzawa Y, Zhivotovsky B, Funahashi T, Cao Y (2004) Adiponectin-induced antiangiogenesis and antitumor activity involve caspase-mediated endothelial cell apoptosis. Proc Natl Acad Sci U S A 101:2476–2481

    Article  CAS  PubMed  Google Scholar 

  50. Crandall DL, Goldstein BM, Huggins F, Cervoni P (1984) Adipocyte blood flow: influence of age, anatomic location, and dietary manipulation. Am J Physiol 247:R46–R51

    CAS  PubMed  Google Scholar 

  51. Cao R, Brakenhielm E, Pawliuk R, Wariaro D, Post MJ, Wahlberg E, Leboulch P, Cao Y (2003) Angiogenic synergism, vascular stability and improvement of hind-limb ischemia by a combination of PDGF-BB and FGF-2. Nat Med 9:604–613

    Article  CAS  PubMed  Google Scholar 

  52. Ohta Y, Ichimura K (2000) Proliferation markers, proliferating cell nuclear antigen, Ki67: 5-bromo-2̃-deoxyuridine, and cyclin D1 in mouse olfactory epithelium. Ann Otol Rhinol Laryngol 109:1046–1048

    CAS  PubMed  Google Scholar 

  53. Yu CC, Woods AL, Levison DA (1992) The assessment of cellular proliferation by immuno-histochemistry: a review of currently available methods and their applications. Histochem J 24:121–131

    Article  CAS  PubMed  Google Scholar 

  54. Ye W, Mairet-Coello G, Dicicco-Bloom E (2006) DNAse I pre-treatment markedly enhances detection of nuclear cyclin-dependent kinase inhibitor p57Kip2 and BrdU double immunos-taining in embryonic rat brain. Histochem Cell Biol 127:195–203

    Article  PubMed  Google Scholar 

  55. Ellwart J, Dormer P (1985) Effect of 5-fluoro-2′-deoxyuridine (FdUrd) on 5-bromo-2′-deoxy-uridine (BrdUrd) incorporation into DNA measured with a monoclonal BrdUrd antibody and by the BrdUrd/Hoechst quenching effect. Cytometry 6:513–520

    Article  CAS  PubMed  Google Scholar 

  56. Gonchoroff NJ, Greipp PR, Kyle RA, Katzmann JA (1985) A monoclonal antibody reactive with 5-bromo-2-deoxyuridine that does not require DNA denaturation. Cytometry 6:506–512

    Article  CAS  PubMed  Google Scholar 

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Brakenhielm, E., Cao, Y. (2008). Angiogenesis in Adipose Tissue. In: Yang, K. (eds) Adipose Tissue Protocols. Methods in Molecular Biology™, vol 456. Humana Press. https://doi.org/10.1007/978-1-59745-245-8_5

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  • DOI: https://doi.org/10.1007/978-1-59745-245-8_5

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-916-1

  • Online ISBN: 978-1-59745-245-8

  • eBook Packages: Springer Protocols

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