Skip to main content
SpringerLink
Log in

Impaired expression of the mitochondrial calcium uniporter suppresses mast cell degranulation

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Calcium ion (Ca2+) uptake into the mitochondrial matrix influences ATP production, Ca2+ homeostasis, and apoptosis regulation. Ca2+ uptake across the ion-impermeable inner mitochondrial membrane is mediated by the mitochondrial Ca2+ uniporter (MCU) complex. The MCU complex forms a pore structure composed of several proteins. MCU is a Ca2+-selective channel in the inner-mitochondrial membrane that allows electrophoretic Ca2+ entry into the matrix. Mitochondrial Ca2+ uptake 1 (MICU1) functions as a Ca2+-sensing regulator of the MCU complex. Previously, by microscopic analysis at the single-cell level, we found that during mast cell activation, mitochondria capture cytosolic Ca2+ in two steps. Consequently, mitochondrial Ca2+ uptake likely plays a role in cellular function through cytosolic Ca2+ buffering. Here, we investigate the role of MCU and MICU1 in mitochondrial Ca2+ uptake and mast cell degranulation using MCU- and MICU1-knockdown (KD) mast cells. Whereas MCU- and MICU1-KD mast cells show normal proliferation rates and mitochondrial membrane potential, they exhibit slow and reduced cytosolic and mitochondrial Ca2+ elevation after antigen stimulation. Moreover, β-hexosaminidase release induced by antigen was significantly suppressed in MCU-KD cells but not MICU1-KD cells. This suggests that both MCU and MICU1 are involved in mitochondrial Ca2+ uptake in mast cells, while MCU plays a role in mast cell degranulation.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Williams GS, Boyman L, Chikando AC, Khairallah RJ, Lederer WJ (2013) Mitochondrial calcium uptake. Proc Natl Acad Sci USA 110:10479–10486

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  2. Perocchi F, Gohil VM, Girgis HS, Bao XR, McCombs JE, Palmer AE, Mootha VK (2010) MICU1 encodes a mitochondrial EF hand protein required for Ca2+ uptake. Nature 467:291–296

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  3. Mallilankaraman K, Doonan P, Cárdenas C, Chandramoorthy HC, Müller M, Miller R et al (2012) MICU1 is an essential gatekeeper for MCU-mediated mitochondrial Ca2+ uptake that regulates cell survival. Cell 151:630–644

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  4. De Stefani D, Raffaello A, Teardo E, Szabò I, Rizzuto R (2011) A forty-kilodalton protein of the inner membrane is the mitochondrial calcium uniporter. Nature 476:336–340

    Article  PubMed Central  PubMed  Google Scholar 

  5. Baughman JM, Perocchi F, Girgis HS, Plovanich M, Belcher-Timme CA, Sancak Y et al (2011) Integrative genomics identifies MCU as an essential component of the mitochondrial calcium uniporter. Nature 476:341–345

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  6. Galli SJ, Kalesnikoff J, Grimbaldeston MA, Piliponsky AM, Williams CMM, Tsai M (2005) Mast cells as “tunable” effector and immunoregulatory cells: recent advances. Annu Rev Immunol 23:749–786

    Article  CAS  PubMed  Google Scholar 

  7. Rivera J (2002) Molecular adapters in FcεRI signaling and the allergic response. Curr Opin Immunol 14:688–693

    Article  CAS  PubMed  Google Scholar 

  8. Calloway N, Vig M, Kinet JP, Holowka D, Baird B (2009) Molecular clustering of STIM1 with Orai1/CRACM1 at the plasma membrane depends dynamically on depletion of Ca2+ stores and on electrostatic interactions. Mol Biol Cell 20:389–399

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  9. Drago I, De Stefani D, Rizzuto R, Pozzan T (2012) Mitochondrial Ca2+ uptake contributes to buffering cytoplasmic Ca2+ peaks in cardiomyocytes. Proc Natl Acad Sci USA 109:12986–12991

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  10. Qiu J, Tan YW, Hagenston AM, Martel MA, Kneisel N, Skehel PA et al (2013) Mitochondrial calcium uniporter Mcu controls excitotoxicity and is transcriptionally repressed by neuroprotective nuclear calcium signals. Nat Commun 4:2034. doi:10.1038/ncomms3034

    PubMed Central  PubMed  Google Scholar 

  11. Alam MR, Groschner LN, Parichatikanond W, Kuo L, Bondarenko AI, Rost R et al (2012) Mitochondrial Ca2+ uptake 1 (MICU1) and mitochondrial Ca2+ uniporter (MCU) contribute to metabolism-secretion coupling in clonal pancreatic β-cells. J Biol Chem 287:34445–34454

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  12. Takekawa M, Furuno T, Hirashima N, Nakanishi M (2012) Mitochondria take up Ca2+ in two steps dependently on store-operated Ca2+ entry in mast cells. Biol Pharm Bull 35:1354–1360

    Article  CAS  PubMed  Google Scholar 

  13. Furuno T, Hirashima N, Onizawa S, Sagiya N, Nakanishi M (2001) Nuclear shuttling of mitogen-activated protein (MAP) kinase (extracellular signal-regulated kinase (ERK) 2) was dynamically controlled by MAP/ERK kinase after antigen stimulation in RBL-2H3 cells. J Immunol 166:4416–4421

    Article  CAS  PubMed  Google Scholar 

  14. Cossarizza A, Baccarani-Contri M, Kalashnikova G, Franceschi C (1993) A new method for the cytofluorimetric analysis of mitochondrial membrane potential using the J-aggregate forming lipophilic cation 5,5’,6,6’-tetrachloro-1,1’,3,3’-tetraethylbenzimidazolcarbocyanine iodide (JC-1). Biochem Biophys Res Commun 197:40–45

    Article  CAS  PubMed  Google Scholar 

  15. Nakamura R, Furuno T, Nakanishi M (2006) The plasma membrane shuttling of CAPRI is related to regulation of mast cell activation. Biochem Biophys Res Commun 347:363–368

    Article  CAS  PubMed  Google Scholar 

  16. Mallilankaraman K, Doonan P, Cárdenas C, Chandramoorthy HC, Müller M, Miller R et al (2012) MICU1 is an essential gatekeeper for MCU-mediated mitochondrial Ca2+ uptake that regulates cell survival. Cell 151:630–644

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  17. Kamer KJ, Sancak Y, Mootha VK (2014) The uniporter: from newly identified parts to function. Biochem Biophys Res Commun 449:370–372

    Article  CAS  PubMed  Google Scholar 

  18. Marchi S, Pinton P (2014) The mitochondrial calcium uniporter complex: molecular components, structure and physiopathological implications. J Physiol 592:829–839

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  19. Pendin D, Greotti E, Pozzan T (2014) The elusive importance of being a mitochondrial Ca2+ uniporter. Cell Calcium 55:139–145

    Article  CAS  PubMed  Google Scholar 

  20. Murgia M, Rizzuto R (2015) Molecular diversity and pleiotropic role of the mitochondrial calcium uniporter. Cell Calcium 58:11–17

    Article  CAS  PubMed  Google Scholar 

  21. Plovanich M, Bogorad RL, Sancak Y, Kamer KJ, Strittmatter L, Li AA et al (2013) MICU2, a paralog of MICU1, resides within the mitochondrial uniporter complex to regulate calcium handling. PLoS ONE 8:e55785. doi:10.1371/journal.pone.0055785

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Raffaello A, De Stefani D, Sabbadin D, Teardo E, Merli G, Picard A et al (2013) The mitochondrial calcium uniporter is a multimer that can include a dominant-negative pore-forming subunit. EMBO J 32:2362–2376

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  23. Mallilankaraman K, Cárdenas C, Doonan PJ, Chandramoorthy HC, Irrinki KM, Golenár T et al (2012) MCUR1 is an essential component of mitochondrial Ca2+ uptake that regulates cellular metabolism. Nat Cell Biol 14:1336–1343

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  24. Sancak Y, Markhard AL, Kitami T, Kovács-Bogdán E, Kamer KJ, Udeshi ND et al (2013) EMRE is an essential component of the mitochondrial calcium uniporter complex. Science 342:1379–1382

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  25. Pan X, Liu J, Nguyen T, Liu C, Sun J, Teng Y et al (2013) The physiological role of mitochondrial calcium revealed by mice lacking the mitochondrial calcium uniporter. Nat Cell Biol 15:1464–1472

    Article  CAS  PubMed  Google Scholar 

  26. Logan CV, Szabadkai G, Sharpe JA, Parry DA, Torelli S, Childs AM et al (2014) Loss-of-function mutations in MICU1 cause a brain and muscle disorder linked to primary alterations in mitochondrial calcium signaling. Nat Genet 46:188–193

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This research was supported, in part, by the Ministry of Education, Culture, Sports, and Technology of Japan (grant nos. 26440083 to T.F.; and 24590060 to M.N.).

Author information

Authors and Affiliations

  1. School of Pharmacy, Aichi Gakuin University, 1-100 Kusumoto-cho, Chikusa-ku, Nagoya, 464-8650, Japan

    Tadahide Furuno, Narumi Shinkai, Yoshikazu Inoh & Mamoru Nakanishi

Authors
  1. Tadahide Furuno
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Narumi Shinkai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yoshikazu Inoh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mamoru Nakanishi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tadahide Furuno.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Furuno, T., Shinkai, N., Inoh, Y. et al. Impaired expression of the mitochondrial calcium uniporter suppresses mast cell degranulation. Mol Cell Biochem 410, 215–221 (2015). https://doi.org/10.1007/s11010-015-2554-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-015-2554-4

Keywords

Search

Navigation