Calmodulin spatial dynamics in RBL-2H3 mast cells

Maria Psatha, Anna Koffer, Muriel Erent, Stephen E Moss, Stephen Bolsover

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Abstract

A line of rat basophilic leukaemia (RBL) cells, a model of mast cells, stably expressing EGFP-tagged calmodulin secreted normally in response to standard agonists. As reported for other cell types, calmodulin was concentrated in the mitotic spindle poles of dividing cells. In unstimulated interphase cells calmodulin was concentrated in the cell cortex and at a single central location. Disruption of cortical actin eliminated the concentration of calmodulin at the cortex while the central calmodulin concentration was associated with an enrichment of tubulin and is likely to represent the centrosome. Following stimulation with either an agonist that crosslinks Fc receptors or co-application of phorbol ester and a calcium ionophore the interior of the cells lost calmodulin while cortical fluorescence became more pronounced but also less uniform. After stimulation discrete bright puncta of calmodulin-EGFP (CaM-EGFP) appeared in the cell interior. Puncta colocalised with moving lysotracker-labelled granules, suggesting that calmodulin may play a role in organising their transport. Our results show that in interphase RBL cells a large fraction of the calmodulin pool is associated with targets in the actin cytoskeleton and demonstrate the utility of this model system for studying calmodulin biology.
Original languageEnglish
Pages (from-to)51-59
Number of pages9
JournalCell Calcium
Volume36
Issue number1
Early online date19 Feb 2004
DOIs
Publication statusPublished - 1 Jul 2004

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Keywords

  • Activins
  • Animals
  • Calmodulin
  • Cell Line, Tumor
  • Cytoskeleton
  • Green Fluorescent Proteins
  • Mast Cells
  • Population Density
  • Population Dynamics
  • Rats
  • Recombinant Fusion Proteins

Cite this

Psatha, M., Koffer, A., Erent, M., Moss, S. E., & Bolsover, S. (2004). Calmodulin spatial dynamics in RBL-2H3 mast cells. Cell Calcium, 36(1), 51-59. https://doi.org/10.1016/j.ceca.2003.11.009