Skip to main content

Adipose Organ Nerves Revealed by Immunohistochemistry#

  • Protocol
Adipose Tissue Protocols

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

Summary

Brown and white adipose tissue have recently gained prominence as key players in obesity and related health problems, such as type-2 diabetes and cardiovascular disease. Brown adipose tissue-dependent nonshivering thermogenesis significantly affects the body's energy balance. Originally considered as a passive store of lipids, white adipose tissue has recently been found to secrete a number of hormones and cytokines and to be thus involved in the control of body metabolism and energy balance at multiple sites. These findings have renewed the interest in adipose organ biology, including its innervation by the autonomic nervous system and sensory nerves. Here, we describe our protocols for detecting different types of adipose tissue nerves by light microscopy using peroxidase immunostaining and by laser scanning confocal microscopy using immunofluorescence. With these techniques, the presence, distribution, and colocalization of autonomic and sensory nerves can be effectively investigated in subcutaneous and visceral adipose depots of normal and obese animals.

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 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
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. Cinti S (1999) The adipose organ, Editrice Kurtis, Milan, Italy

    Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  4. Jaworski K, Sarkadi-Nagy E, Duncan R, Ahmadian M, Sul HS (2007) Regulation of triglyceride metabolism. IV. Hormonal regulation in lipolysis in adipose tissue. Am J Physiol Gastrointest Liver Physiol 293:G1–4

    Article  CAS  PubMed  Google Scholar 

  5. Bartness TJ, Kay Song C, Shi H, Bowers RR, Foster MT (2005) Brain-adipose tissue cross talk. Proc Nutr Soc 64:53–64

    Article  CAS  PubMed  Google Scholar 

  6. Bartness TJ, Bamshad M (1998) Innervation of mammalian white adipose tissue: implications for the regulation of total body fat. Am J Physiol 275:R1399–1411

    CAS  PubMed  Google Scholar 

  7. Giordano A, Morroni M, Carle F, Gesuita R, Marchesi GF, Cinti S (1998) Sensory nerves affect the recruitment and differentiation of rat periovarian brown adipocytes during cold acclimation. J Cell Sci 111:2587–2594

    CAS  PubMed  Google Scholar 

  8. Slavin BG, Ballard KW (1978) Morphological studies on the adrenergic innervation of white adipose tissue. Anat Rec 191:377–389

    Article  CAS  PubMed  Google Scholar 

  9. Giordano A, Morroni M, Santone G, Marchesi GF, Cinti S (1996) Tyrosine hydroxylase, neuropeptide Y, substance P, calcitonin gene-related peptide and vasoactive intestinal peptide in nerves of rat periovarian adipose tissue: an immunohistochemical and ultrastructural investigation. J Neurocytol 25:125–136

    Article  CAS  PubMed  Google Scholar 

  10. Norman D, Mukherjee S, Symons D, Jung RT, Lever JD (1988) Neuropeptides in interscapular and perirenal brown adipose tissue in the rat: a plurality of innervation. J. Neurocytol 17:305–311

    Article  CAS  PubMed  Google Scholar 

  11. De Matteis R, Ricquier D, Cinti S (1998) TH-, NPY-, SP-, and CGRP-immunoreactive nerves in interscapular brown adipose tissue of adult rats acclimated at different temperatures: an immunohistochemical study. J Neurocytol 27:877–886

    Article  PubMed  Google Scholar 

  12. Giordano A, Frontini A, Castellucci M, Cinti S (2004) Presence and distribution of cholinergic nerves in rat mediastinal brown adipose tissue. J Histochem Cytochem 52:923–930

    Article  CAS  PubMed  Google Scholar 

  13. Giordano A, Frontini A, Murano I, Tonello C, Marino MA, Carruba MO, Nisoli E, Cinti S (2005) Regional-dependent increase of sympathetic innervation in rat white adipose tissue during prolonged fasting. J Histochem Cytochem 53:679–687

    Article  CAS  PubMed  Google Scholar 

  14. Giordano A, Song CK, Bowers RR, Ehlen JC, Frontini A, Cinti S, Bartness TJ (2006) White adipose tissue lacks significant vagal innervation and immunohistochemical evidence of para-sympathetic innervation. Am J Physiol Regul Int Comp Physiol 291:R1243–R1255

    CAS  Google Scholar 

  15. Hsu SM, Raine L, Fanger H (1981) Use of avidin-biotin-peroxidase complex (ABC) in immu-noperoxidase technique: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem 29:577–580

    CAS  PubMed  Google Scholar 

  16. Cinti S, Zingaretti MC, Cancello R, Ceresi E, Ferrara P (2001) Morphologic techniques for the study of brown adipose tissue and white adipose tissue. Methods Mol Biol. 155:21–51

    CAS  PubMed  Google Scholar 

  17. Giordano A, Centemeri C, Zingaretti MC, Cinti S (2002) Sibutramine-dependent brown fat activation in rats: an immunohistochemical study. Int J Obesity Relat Metab Disord 26:354–360

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was financed by grants from Marche Polytechnic University (Contributi Ricerca Scientifica) and the Ministry of University (FIRB Internazionalizzazione RBIN047PZY, 2005).

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2008 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Giordano, A., Frontini, A., Cinti, S. (2008). Adipose Organ Nerves Revealed by Immunohistochemistry#. 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_6

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-245-8_6

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-916-1

  • Online ISBN: 978-1-59745-245-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics