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The Use of Cre/loxP Inducible Mouse Models to Dissect the Specific Roles of Calcineurin Signaling in Myeloid Cells

  • Teresa Zelante
  • Jan FričEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1929)

Abstract

The calcineurin/nuclear factor of activated T-cell (NFAT) signaling pathway is important for the development and function of different leukocyte subsets. Calcineurin is activated through increased intracellular levels of calcium that is triggered by cell receptors. The phosphatase activity of calcineurin dephosporylates the members of NFAT transcription factor family and activates their translocation to the nucleus and further starts the transcription of NFAT-regulated genes. Although calcineurin is expressed ubiquitously, its functions are cell- and tissue-specific. The genetically engineered murine models allow to dissect the calcineurin roles in tissue-specific manner and as such have been essential to pursue the understanding of important role of calcineurin/NFAT pathway in orchestrating the developmental and immune response processes. This protocol describes several examples of the use of Cre/loxP inducible mouse models deficient in calcineurin in different myeloid subsets to dissect the cell- and tissue-specific roles of calcineurin signaling.

Key words

Cre/lox breeding Dendritic cells Calcineurin Calcium Macrophages Granulocytes NFAT Myeloid cells 

Notes

Acknowledgments

Supported by European Social Fund and European Regional Development Fund, Project MAGNET (No. CZ.02.1.01/0.0/0.0/15_003/0000492), and by the Italian Grant “Programma per Giovani Ricercatori - Rita Levi Montalcini 2013”.

References

  1. 1.
    Fric J, Lim CX, Mertes A, Lee BT, Vigano E, Chen J, Zolezzi F, Poidinger M, Larbi A, Strobl H, Zelante T, Ricciardi-Castagnoli P (2014) Calcium and calcineurin-NFAT signaling regulate granulocyte-monocyte progenitor cell cycle via Flt3-L. Stem Cells 32:3232–3244CrossRefGoogle Scholar
  2. 2.
    Zelante T, Wong AY, Ping TJ, Chen J, Sumatoh HR, Vigano E, Hong Bing Y, Lee B, Zolezzi F, Fric J, Newell EW, Mortellaro A, Poidinger M, Puccetti P, Ricciardi-Castagnoli P (2015) CD103(+) dendritic cells control Th17 cell function in the lung. Cell Rep 12:1789–1801CrossRefGoogle Scholar
  3. 3.
    Herbst S, Shah A, Mazon Moya M, Marzola V, Jensen B, Reed A, Birrell MA, Saijo S, Mostowy S, Shaunak S, Armstrong-James D (2015) Phagocytosis-dependent activation of a TLR9-BTK-calcineurin-NFAT pathway co-ordinates innate immunity to Aspergillus fumigatus. EMBO Mol Med 7:240–258CrossRefGoogle Scholar
  4. 4.
    Zelante T, Wong AY, Mencarelli A, Foo S, Zolezzi F, Lee B, Poidinger M, Ricciardi-Castagnoli P, Fric J (2017) Impaired calcineurin signaling in myeloid cells results in downregulation of pentraxin-3 and increased susceptibility to aspergillosis. Mucosal Immunol 10:470–480CrossRefGoogle Scholar
  5. 5.
    Zeng H, Chattarji S, Barbarosie M, Rondi-Reig L, Philpot BD, Miyakawa T, Bear MF, Tonegawa S (2001) Forebrain-specific calcineurin knockout selectively impairs bidirectional synaptic plasticity and working/episodic-like memory. Cell 107:617–629CrossRefGoogle Scholar
  6. 6.
    Neilson JR, Winslow MM, Hur EM, Crabtree GR (2004) Calcineurin B1 is essential for positive but not negative selection during thymocyte development. Immunity 20:255–266CrossRefGoogle Scholar
  7. 7.
    Caton ML, Smith-Raska MR, Reizis B (2007) Notch-RBP-J signaling controls the homeostasis of CD8- dendritic cells in the spleen. J Exp Med 204:1653–1664CrossRefGoogle Scholar
  8. 8.
    Clausen BE, Burkhardt C, Reith W, Renkawitz R, Forster I (1999) Conditional gene targeting in macrophages and granulocytes using LysMcre mice. Transgenic Res 8:265–277CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Experimental MedicineUniversity of PerugiaPerugiaItaly
  2. 2.Center for Translational Medicine, International Clinical Research Center, St. Anne’s University Hospital BrnoBrnoCzech Republic

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