Micro-scale flow cytometry-based and biochemical analysis of lipid signaling in primary B cell subpopulations
- 24 December 2007
- journal article
- Published by Springer Science and Business Media LLC
- Vol. 9, 73-83
- https://doi.org/10.1251/bpo135
Abstract
B cell subpopulations in the spleen have been extensively characterized phenotypically; however, biochemical properties of these cell populations following B cell antigen receptor engagement have not been fully determined due to technical difficulties and limiting cell numbers. We therefore employed mini-scale protocols to assess lipid signaling, particularly that of diacylglycerol and inositol trisphosphate, with as few as 0.5x106 purified early (T1) and late (T2) transitional B cells. Additionally, utilizing flow cytometric techniques, we determined levels of phosphatidylinositol bisphosphate and calcium mobilization in T1 and T2 cells, as well as mature follicular and marginal zone B cells using less than 1x106 primary B cells. Thus, these biochemical and flow cytometric methodologies can be used to analyse signal-induced changes in phosphatidylinositol bisphosphate levels, diacylglycerol and inositol triphosphate production and calcium in each B cell population.This publication has 25 references indexed in Scilit:
- Inositol 1,3,4,5-tetrakisphosphate controls proapoptotic Bim gene expression and survival in B cellsProceedings of the National Academy of Sciences of the United States of America, 2007
- Characterization of marginal zone B cell precursorsThe Journal of Experimental Medicine, 2005
- BTK Regulates PtdIns-4,5-P2 Synthesis: Importance for Calcium Signaling and PI3K ActivityImmunity, 2003
- B Cell receptor directs the activation of NFAT and NF-κB via distinct molecular mechanismsExperimental Cell Research, 2003
- Coupling Between B Cell Receptor and Phospholipase C-γ2 Is Essential for Mature B Cell DevelopmentThe Journal of Experimental Medicine, 2003
- Transitional Type 1 and 2 B Lymphocyte Subsets Are Differentially Responsive to Antigen Receptor SignalingOnline Journal of Public Health Informatics, 2002
- Biochemical and Functional Characterization of Inositol 1,3,4,5,6-Pentakisphosphate 2-KinasesOnline Journal of Public Health Informatics, 2000
- A Phospholipase C-Dependent Inositol Polyphosphate Kinase Pathway Required for Efficient Messenger RNA ExportScience, 1999
- Defective B cell development and function in Btk-deficient miceImmunity, 1995
- B lymphocyte receptors and polyphosphoinositide degradationCell, 1985