Skip to main content

The Effect of Macrophage Migration Inhibitory Factor on Intestinal Permeability: FITC-Dextran Serum Measurement and Transmission Electron Microscopy

  • Protocol
  • First Online:
Book cover Macrophage Migration Inhibitory Factor

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2080))

Abstract

Macrophage migration inhibitory factor (MIF) is a molecule with multiple functions: from enforcing the immune system to fight bacterial infection to the regulation of insulin activity. Also, MIF is expressed by enterocytes that line the intestinal border toward the lumen, and in M cells, where it regulates phagocytosis of antigens from the lumen of the gut and their transport to Peyer’s patches. Since there were no data on the role of MIF in the maintenance of the intestinal barrier, we used MIF-deficient mice bred on C57BL/6 background as a model for the investigation of intestinal permeability. The obtained results indicate that the absence of MIF increases intestinal permeability. Here we describe two methods for measuring intestinal permeability in mice: detection of orally delivered FITC-dextran in the serum and transmission electron microscopy used for visualization and measurement of cell-to-cell connections width.

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 119.99
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. Calandra T, Roger T (2003) Macrophage migration inhibitory factor: a regulator of innate immunity. Nat Rev Immunol 3:791–800

    Article  CAS  Google Scholar 

  2. Stojanovic I, Mirkov I, Kataranovski M et al (2011) A role for macrophage migration inhibitory factor in protective immunity against Aspergillus fumigatus. Immunobiology 216:1018–1027

    Article  CAS  Google Scholar 

  3. Stojanovic I, Cvjeticanin T, Lazaroski S et al (2009) Macrophage migration inhibitory factor stimulates interleukin-17 expression and production in lymph node cells. Immunology 126:74–83

    Article  CAS  Google Scholar 

  4. Gao XM, Liu Y, White D et al (2011) Deletion of macrophage migration inhibitory factor protects the heart from severe ischemia-reperfusion injury: a predominant role of anti-inflammation. J Mol Cell Cardiol 50:991–999

    Article  CAS  Google Scholar 

  5. Flaster H, Bernhagen J, Calandra T et al (2007) The macrophage migration inhibitory factor-glucocorticoid dyad: regulation of inflammation and immunity. Mol Endocrinol 21:1267–1280

    Article  CAS  Google Scholar 

  6. Waeber G, Calandra T, Roduit R et al (1997) Insulin secretion is regulated by the glucose-dependent production of islet beta cell macrophage migration inhibitory factor. Proc Natl Acad Sci U S A 94:4782–4787

    Article  CAS  Google Scholar 

  7. Serre-Beinier V, Toso C, Morel P et al (2010) Macrophage migration inhibitory factor deficiency leads to age-dependent impairment of glucose homeostasis in mice. J Endocrinol 206:297–306

    Article  CAS  Google Scholar 

  8. Nikolic I, Vujicic M, Saksida T et al (2013) The role of endogenous glucocorticoids in glucose metabolism and immune status of MIF-deficient mice. Eur J Pharmacol 714(1–3):498–506

    Article  CAS  Google Scholar 

  9. Vujicic M, Senerovic L, Nikolic I et al (2014) The critical role of macrophage migration inhibitory factor in insulin activity. Cytokine 69(1):39–46

    Article  CAS  Google Scholar 

  10. Ohkawara T, Nishihira J, Takeda H et al (2002) Amelioration of dextran sulfate sodium-induced colitis by anti-macrophage migration inhibitory factor antibody in mice. Gastroenterology 123:256–270

    Article  CAS  Google Scholar 

  11. Ohkawara T, Mitsuyama K, Takeda H et al (2008) Lack of macrophage migration inhibitory factor suppresses innate immune response in murine dextran sulfate sodium-induced colitis. Scand J Gastroenterol 43:1497–1504

    Article  CAS  Google Scholar 

  12. Man AL, Lodi F, Bertelli E et al (2008) Macrophage migration inhibitory factor plays a role in the regulation of microfold (M) cell-mediated transport in the gut. J Immunol 181:5673–5680

    Article  CAS  Google Scholar 

  13. Tlaskalová-Hogenová H, Stepánková R, Hudcovic T et al (2004) Commensal bacteria (normal microflora), mucosal immunity and chronic inflammatory and autoimmune diseases. Immunol Lett 93:97–108

    Article  Google Scholar 

  14. Vujicic M, Saksida T, Despotovic S et al (2018) The role of macrophage migration inhibitory factor in the function of intestinal barrier. Sci Rep 8(1):6337

    Article  Google Scholar 

  15. Yan Y, Kolachala V, Dalmasso G et al (2009) Temporal and spatial analysis of clinical and molecular parameters in dextran sodium sulfate induced colitis. PLoS One 4(6):e6073

    Article  Google Scholar 

  16. Martinovic T, Ciric D, Pantic I et al (2018) Unusual shape and structure of lymphocyte nuclei is linked to hyperglycemia in type 2 diabetes patients. Tissue Cell 52:92–100

    Article  CAS  Google Scholar 

  17. Graham L, Orenstein JM (2007) Processing tissue and cells for transmission electron microscopy in diagnostic pathology and research. Nat Protoc 2(10):2439–2450

    Article  CAS  Google Scholar 

  18. Schindelin J, Arganda-Carreras I, Frise E et al (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9(7):676–682

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia (grant 173013 and 175005).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Ivana Stojanović .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Vujičić, M., Despotović, S., Saksida, T., Stojanović, I. (2020). The Effect of Macrophage Migration Inhibitory Factor on Intestinal Permeability: FITC-Dextran Serum Measurement and Transmission Electron Microscopy. In: Harris, J., Morand, E. (eds) Macrophage Migration Inhibitory Factor. Methods in Molecular Biology, vol 2080. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9936-1_17

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9936-1_17

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9935-4

  • Online ISBN: 978-1-4939-9936-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics