Ratiometric analysis of calcium mobilization

Abstract One of the earliest events in lymphocyte activation is an increase in the concentration of cytosolic free Ca 2 +. Antigen recognition, chemotaxis, apoptosis and cell cycle progression are all associated with changes in cytosolic free Ca 2 + levels. The kinetics, subcellular localization, and degree of Ca 2 + mobilization can be assessed with fluorescent Ca 2 + indicators. Ratiometric analysis of Ca 2 + mobilization reduces errors due to variations in cell size and shape, indicator loading and instrument variability that can otherwise limit the usefulness of fluorescent Ca 2 + indicators. The purposes of this paper are to summarize key characteristics of some useful Ca 2 +-sensitive fluorescent dyes (Indo-1, Fura-2, Fluo-3, Fluo-4 and Fura Red) and to review published methods for the optimization of Ca 2 + mobilization analysis with ratiometric indicators. Analysis of Ca 2 + mobilization provides a rapid, quantitative readout of lymphocyte activation.

[1]  B. Herman,et al.  Assessment of Fura-2 for measurements of cytosolic free calcium. , 1990, Cell calcium.

[2]  Peter Lipp,et al.  Ratiometric confocal Ca2+-measurements with visible wavelength indicators in isolated cardiac myocytes , 1993 .

[3]  G. Hajnóczky,et al.  Mitochondria Suppress Local Feedback Activation of Inositol 1,4,5-Trisphosphate Receptors by Ca2+ * , 1999, The Journal of Biological Chemistry.

[4]  B. Stokes,et al.  Quantitation of intracellular free calcium in single adult cardiomyocytes by fura-2 fluorescence microscopy: calibration of fura-2 ratios. , 1987, Biochemical and biophysical research communications.

[5]  D. Plas,et al.  Expression of dominant-negative src-homology domain 2-containing protein tyrosine phosphatase-1 results in increased Syk tyrosine kinase activity and B cell activation. , 1999, Journal of immunology.

[6]  Peter Lipp,et al.  Calcium - a life and death signal , 1998, Nature.

[7]  P. Rabinovitch,et al.  Chapter 10 Intracellular Ionized Calcium , 1994 .

[8]  P. Rabinovitch,et al.  Heterogeneity among T cells in intracellular free calcium responses after mitogen stimulation with PHA or anti-CD3. Simultaneous use of indo-1 and immunofluorescence with flow cytometry. , 1986, Journal of immunology.

[9]  E. Gruenstein,et al.  Intracellular Ca2+ measurement with Indo-1 in substrate-attached cells: advantages and special considerations. , 1990, Cell calcium.

[10]  P. Rabinovitch,et al.  Measurement of Intracellular Calcium Ions by Flow Cytometry , 1997, Current protocols in cytometry.

[11]  G. Klaus,et al.  Cross-linking of surface immunoglobulin on B lymphocytes induces both intracellular Ca2+ release and Ca2+ influx: analysis with indo-1. , 1986, Biochemical and biophysical research communications.

[12]  A. Guse.  Ca2+ signaling in T-lymphocytes. , 1998, Critical reviews in immunology.

[13]  R. Tsien,et al.  A new generation of Ca2+ indicators with greatly improved fluorescence properties. , 1985, The Journal of biological chemistry.

[14]  J. Brugge,et al.  Leukocyte protein tyrosine kinases: potential targets for drug discovery. , 1997, Annual review of immunology.

[15]  R. Tsien,et al.  Active involvement of Ca2+ in mitotic progression of Swiss 3T3 fibroblasts , 1990, The Journal of cell biology.

[16]  Martin D. Bootman,et al.  The elemental principles of calcium signaling , 1995, Cell.

[17]  R. Tsien.  A non-disruptive technique for loading calcium buffers and indicators into cells , 1981, Nature.

[18]  R. Silver,et al.  Ratio imaging: practical considerations for measuring intracellular calcium and pH in living tissue. , 1998, Methods in cell biology.

[19]  P. Rabinovitch,et al.  Improved sensitivity in flow cytometric intracellular ionized calcium measurement using fluo-3/Fura Red fluorescence ratios. , 1994, Cytometry.

[20]  J. Putney.  Intimate Plasma Membrane–ER Interactions Underlie Capacitative Calcium Entry: “Kissin' Cousins” , 1999, Cell.

[21]  L. Santella.  The role of calcium in the cell cycle: facts and hypotheses. , 1998, Biochemical and biophysical research communications.

[22]  Christopher C. Goodnow,et al.  Differential activation of transcription factors induced by Ca2+ response amplitude and duration , 1997, Nature.

[23]  R. Tsien,et al.  Flow cytometric analysis of murine splenic B lymphocyte cytosolic free calcium response to anti-IgM and anti-IgD. , 1987, Cytometry.

[24]  Lawrence M. Lifshitz,et al.  Close contacts with the endoplasmic reticulum as determinants of mitochondrial Ca2+ responses. , 1998, Science.

[25]  M. Poenie,et al.  New fluorescent calcium indicators designed for cytosolic retention or measuring calcium near membranes. , 1995, Biophysical journal.

[26]  S. Orrenius,et al.  The role of calcium in apoptosis. , 1998, Cell calcium.

[27]  A. DeFranco,et al.  The complexity of signaling pathways activated by the BCR. , 1997, Current opinion in immunology.

[28]  A Miyawaki,et al.  Dynamic and quantitative Ca2+ measurements using improved cameleons. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[29]  A. Griffioen,et al.  Improved method for measuring intracellular Ca++ with fluo-3. , 1990, Cytometry.

[30]  J. Kao,et al.  Practical aspects of measuring [Ca2+] with fluorescent indicators. , 1994, Methods in cell biology.

[31]  R Y Tsien,et al.  Calcium homeostasis in intact lymphocytes: cytoplasmic free calcium monitored with a new, intracellularly trapped fluorescent indicator , 1982, The Journal of cell biology.

[32]  Simon C Watkins,et al.  Current Protocols In Cytometry , 1997 .

[33]  P. Negulescu,et al.  Polarity of T cell shape, motility, and sensitivity to antigen. , 1996, Immunity.

[34]  A. Basile,et al.  Skeletal Muscle Type Ryanodine Receptor Is Involved in Calcium Signaling in Human B Lymphocytes* , 1999, The Journal of Biological Chemistry.

[35]  R. Floto,et al.  IgG-induced Ca2+ oscillations in differentiated U937 cells; a study using laser scanning confocal microscopy and co-loaded fluo-3 and fura-red fluorescent probes. , 1995, Cell calcium.

[36]  A. Trautmann,et al.  Antigen recognition by helper T cells elicits a sequence of distinct changes of their shape and intracellular calcium , 1994, Current Biology.

[37]  R. Tsien,et al.  Fluorescent indicators for cytosolic calcium based on rhodamine and fluorescein chromophores. , 1989, The Journal of biological chemistry.

[38]  Richard P. Haugland,et al.  Handbook of fluorescent probes and research chemicals , 1996 .

[39]  A. W. Simpson.  Fluorescent Measurement of [Ca 2+ ] c , 1999 .