Skip to main content

Analysis of cGMP Signaling in Adipocytes

  • Protocol
  • First Online:
Guanylate Cyclase and Cyclic GMP

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1020))

Abstract

Obesity has reached pandemic dimensions with more than half a billion adults affected worldwide. Detailed knowledge of adipose biology is required for the development of urgently needed novel therapies directed against obesity. Two types of adipose tissue can be distinguished in humans and mice: white adipose tissue (WAT), which primarily stores energy in the form of lipids and has endocrine functions. In contrast, brown adipose tissue (BAT) dissipates energy in the form of heat (thermogenesis). Recent studies in humans demonstrated that BAT not only plays a role for non-shivering thermogenesis in newborns but is also metabolically active in adults. Here, we describe protocols for the generation of cellular models for the analysis of adipogenesis as well as function of brown and white fat. These models are based on the in vitro differentiation of mesenchymal stem cells (MSCs) isolated from adipose tissues. Using specific differentiation protocols, the role of cGMP signaling in both brown as well as white adipocytes can be studied.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Finucane MM, Stevens GA, Cowan MJ et al (2011) National, regional, and global trends in body-mass index since 1980: systematic analysis of health examination surveys and epidemiological studies with 960 country-years and 9.1 million participants. Lancet 377:557–567

    Article  PubMed  Google Scholar 

  2. Ahima RS (2011) Digging deeper into obesity. J Clin Invest 121:2076–2079

    Article  PubMed  CAS  Google Scholar 

  3. Karastergiou K, Mohamed-Ali V (2010) The autocrine and paracrine roles of adipokines. Mol Cell Endocrinol 318:69–78

    Article  PubMed  CAS  Google Scholar 

  4. Wymann MP, Schneiter R (2008) Lipid signalling in disease. Nat Rev Mol Cell Biol 9:162–176

    Article  PubMed  CAS  Google Scholar 

  5. Deng Y, Scherer PE (2010) Adipokines as novel biomarkers and regulators of the ­metabolic syndrome. Ann N Y Acad Sci 1212:E1–E19

    Article  PubMed  Google Scholar 

  6. Cannon B, Nedergaard J (2004) Brown adipose tissue: function and physiological significance. Physiol Rev 84:277–359

    Article  PubMed  CAS  Google Scholar 

  7. Haas B, Mayer P, Jennissen K et al (2009) Protein kinase G controls brown fat cell differentiation and mitochondrial biogenesis. Sci Signal 2:ra78

    Article  PubMed  Google Scholar 

  8. Nikolic DM, Li Y, Liu S et al (2011) Overexpression of constitutively active PKG-I protects female, but not male mice from ­diet-induced obesity. Obesity (Silver Spring) 19:784–791

    Article  CAS  Google Scholar 

  9. Zhang X, Ji J, Yan G et al (2010) Sildenafil promotes adipogenesis through a PKG pathway. Biochem Biophys Res Commun 396:1054–1059

    Article  PubMed  CAS  Google Scholar 

  10. Sengenes C, Bouloumie A, Hauner H et al (2003) Involvement of a cGMP-dependent pathway in the natriuretic peptide-mediated hormone-sensitive lipase phosphorylation in human adipocytes. J Biol Chem 278:48617–48626

    Article  PubMed  CAS  Google Scholar 

  11. Rosen ED, MacDougald OA (2006) Adipocyte differentiation from the inside out. Nat Rev Mol Cell Biol 7:885–896

    Article  PubMed  CAS  Google Scholar 

  12. Nechad M (1983) Development of brown fat cells in monolayer culture. II. Ultrastructural characterization of precursors, differentiating adipocytes and their mitochondria. Exp Cell Res 149:119–127

    Article  PubMed  CAS  Google Scholar 

  13. Ostertag A, Jones A, Rose AJ et al (2010) Control of adipose tissue inflammation through TRB1. Diabetes 59:1991–2000

    Article  PubMed  CAS  Google Scholar 

  14. Rodeheffer MS, Birsoy K, Friedman JM (2008) Identification of white adipocyte progenitor cells in vivo. Cell 135:240–249

    Article  PubMed  CAS  Google Scholar 

  15. Mitschke et al (2013) Increased cGMP ­promotes healthy expansion and browning of white adipose tissue. FASEB J 27(4):1621–1630

    Article  PubMed  CAS  Google Scholar 

  16. Butt E, Abel K, Krieger M et al (1994) cAMP- and cGMP-dependent protein kinase phosphorylation sites of the focal adhesion vasodilator-stimulated phosphoprotein (VASP) in vitro and in intact human platelets. J Biol Chem 269:14509–14517

    PubMed  CAS  Google Scholar 

  17. Seale P, Conroe HM, Estall J et al (2011) Prdm16 determines the thermogenic program of subcutaneous white adipose tissue in mice. J Clin Invest 121:96–105

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

This work was supported by the DFG.

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Jennissen, K., Haas, B., Mitschke, M.M., Siegel, F., Pfeifer, A. (2013). Analysis of cGMP Signaling in Adipocytes. In: Krieg, T., Lukowski, R. (eds) Guanylate Cyclase and Cyclic GMP. Methods in Molecular Biology, vol 1020. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-459-3_11

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-459-3_11

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-458-6

  • Online ISBN: 978-1-62703-459-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics