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Intravital quantification reveals dynamic calcium concentration changes across B cell differentiation stages.
Carolin Ulbricht1,2, Ruth Leben3, Asylkhan Rakhymzhan3
1Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Department of Rheumatology and Clinical Immunology, Charitéplatz 1, Berlin, Germany.
Elife
|March 22, 2021
Summary
Researchers developed a new mouse model to measure calcium levels in immune cells. This allows for precise quantification of calcium dynamics crucial for understanding immune responses and autoimmune diseases.
Area of Science:
- Immunology
- Cell Biology
- Biophysics
Background:
- Calcium ions (Ca2+) act as universal second messengers in eukaryotic cells.
- Calcium signaling is vital for immune cell development, pathogen adaptation, and cellular processes.
- Understanding intracellular calcium dynamics is key to deciphering immune responses and diseases.
Purpose of the Study:
- To introduce a novel Förster resonance energy transfer (FRET)-reporter mouse, YellowCaB, for in vivo calcium imaging in B lymphocytes.
- To develop a new numerical analysis for quantifying absolute cytoplasmic calcium concentrations.
- To investigate the dynamic nature and heterogeneity of calcium levels in activated B cells during affinity maturation.
Main Methods:
- Generation of the YellowCaB reporter mouse expressing the genetically encoded calcium indicator TN-XXL.
- Utilizing two-photon fluorescence lifetime imaging microscopy to measure FRET-donor quenching.
- Application of novel numerical analysis to determine absolute cytoplasmic calcium concentrations in vivo.
Main Results:
- Demonstrated that calcium levels in activated B cells are highly dynamic during affinity maturation.
- Revealed that B cell activation induces persistent heterogeneity in cytoplasmic calcium concentrations.
- Successfully extracted absolute cytosolic calcium concentrations for the first time in activated B cells in vivo.
Conclusions:
- The YellowCaB mouse model provides a powerful tool for real-time, quantitative analysis of calcium signaling in B cells.
- Precise characterization of calcium concentrations is essential for understanding long-term immune responses and autoimmune conditions.
- This study offers new insights into the role of calcium dynamics in immune cell function and adaptation.
