Real-Time Imaging of Ca2+ Mobilization and Degranulation in Mast Cells
Mast cells play a key role in allergy and inflammation processes as part of the immune response. The activation of mast cells via antigen binding and cross-linking of IgE receptors initiates the onset of dramatic calcium (Ca2+) mobilization dynamics that promote the release of mediators of inflammation and allergy. Ca2+ signaling in mast cells has been studied extensively using a variety of research tools and techniques. In these studies, a large number of proteins have been identified to participate in various stages of these processes.
Here we describe single-cell imaging as an important approach for examining Ca2+ signaling and exocytosis in mast cells. Single-cell imaging tools have advanced significantly over the last 10 years, in part due to improvements in microscope technology and in part due to the development of a new generation of Ca2+ indicators and genetically encoded Ca2+ sensors. The single-cell imaging techniques described here provide the spatial and temporal resolution required to decipher the signaling events that are critical for mast cell functions.
Key wordsLive-cell imaging Calcium (Ca2+) signaling Calcium dynamics Mast cell degranulation
- 6.Tian L, Hires SA, Mao T, Huber D, Chiappe ME, Chalasani SH, Petreanu L, Akerboom J, McKinney SA, Schreiter ER, Bargmann CI, Jayaraman V, Svoboda K, Looger LL (2009) Imaging neural activity in worms, flies and mice with improved GCaMP calcium indicators. Nat Methods 6(12):875–881. doi:10.1038/nmeth.1398 PubMedCentralPubMedCrossRefGoogle Scholar
- 16.Gadi D, Wagenknecht-Wiesner A, Holowka D, Baird B (2011) Sequestration of phosphoinositides by mutated MARCKS effector domain inhibits stimulated Ca(2+) mobilization and degranulation in mast cells. Mol Biol Cell 22(24): 4908–4917. doi:10.1091/mbc.E11-07-0614 PubMedCentralPubMedCrossRefGoogle Scholar
- 17.Hohman RJ, Dreskin SC (2001) Measuring degranulation of mast cells. In: John E. Coligan et al (eds). Current protocols in immunology. Chapter 7:Unit 7;26. doi:10.1002/0471142735.im0726s08