← Back to Search
A Non-disruptive Technique For Loading Calcium Buffers And Indicators Into Cells
Published 1981 · Chemistry, Medicine
Save to my Library
Download PDFAnalyze on Scholarcy
Present methods for measuring or buffering intracellular free calcium concentrations are almost entirely limited to robust and well anchored cells which can tolerate insertion of ion-selective microelectrodes or microinjection of calcium indicators or buffers into one cell at a time1. A very few types of small cells can be loaded with buffers or indicators during controlled lysis2–4, but such procedures grossly perturb membrane integrity and soluble cytoplasmic constituents. Liposome fusion releases only trace quantities of the trapped solute into the cytoplasm5,6 and incorporates foreign lipid into the target cell membranes. I now describe a simple technique which loads Ca2+-selective chelators into the cytoplasm of intact cells in suspension and avoids the disadvantages of previous methods. The chelators are made temporarily membrane permeable by masking their four carboxylates with special esterifying groups which then hydrolyse inside the cells, regenerating and trapping the original chelators. The method is demonstrated on red cells, mast cells and lymphocytes.
This paper references
379. Some novel penicillin derivatives
A. Jansen (1965)
Membrane properties of living mammalian cells as studied by enzymatic hydrolysis of fluorogenic esters.
B. Rotman (1966)
Actions of a carbocyanine dye on calcium‐dependent potassium transport in human red cell ghosts.
T. J. Simons (1979)
The preparation of human red cell ghosts containing calcium buffers.
T. J. Simons (1976)
Current Topics in Membranes and Transport
F. Bronner (1970)
Calcium-dependent exocytosis in bovine adrenal medullary cells with leaky plasma membranes
P. Baker (1978)
Intracellular pH measurements in Ehrlich ascites tumor cells utilizing spectroscopic probes generated in situ.
J. A. Thomas (1979)
Chemical modification of crayfish axons by protein crosslinking aldehydes
P. Shrager (1969)
Improved Calcium Ion-Selective Electrode Based on a Neutral Carrier
D. Ammann (1975)
New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures.
R. Tsien (1980)
Interactions of lysophospholipids and mast cells
T. Martin (1979)
Variable Ca sensitivity of a K-selective channel in intact red-cell membranes
V. Lew (1976)
A calcium hypothesis for the control of cell growth.
J. Metcalfe (1980)
Effect of Formaldehyde and Glutaraldehyde on Electrical Properties of Cardiac Purkinje Fibers
H. Fozzard (1969)
Synthese von Fulvenen über 1‐Chloralkyl‐acetate
M. Neuenschwander (1977)
Potential difference and the distribution of ions across the human red blood cell membrane; a study of the mechanism by which the fluorescent cation, diS‐C3‐(5) reports membrane potential.
S. Hladky (1976)
Changes of intracellular Ca++ as measured by arsenazo III in relation to the K permeability of human erythrocyte ghosts.
D. Yingst (1978)
The exogenous lipid requirement for histamine release from rat peritoneal mast cells stimulated by concanavalin A
G. Smith (1979)
Studies on the mechanism by which cyanine dyes measure membrane potential in red blood cells and phosphatidylcholine vesicles.
P. Sims (1974)
Receptors and Recognition
P. Cuatrecasas (1976)
This paper is referenced by
Supervised Neural Network Structure Recovery
I. D. M. Abril (2014)
High-throughput analyses of IP3 receptor behavior.
C. Taylor (2013)
Involvement of calcium in the transient depolarization of E.coli cytoplasmic membrane induced by phage adsorption: a study with the fluorescent calcium indicator QUIN2.
P. Boulanger (1985)
Intracellular free Ca2+ and the hypercontracture of adult rat heart myocytes.
M. R. Lambert (1986)
Ionic channels: II. Voltage‐ and agonist‐gated and agonist‐modified channel properties and structure
R. Ruff (1986)
Cell Movement, Excitability, and Contractility
P. Naccache (1987)
Calcium control of exocytosis in bovine adrenal medullary cells
P. F. Baker (1984)
Optical monitoring of mitochondrial membrane potential in intact myocardium and its application to the development of tissue injury caused by calcium readmission to the isolated perfused heart.
I. Hassinen (1986)
Remodelling of store-operated Ca2+ entry and G0/G1 cell cycle arrest
Marwa Etfat Eliwa (2017)
ION-SELECTIVE MICROELECTRODES AND FLUORESCENT PROBES FOR MEASUREMENT OF INTRACELLULAR ION ACTIVITIES
R. Y. Tsien (1982)
CHAPTER EIGHT – Fluorescent Probes in Practice – Potential Artifacts
J. Hoyland (1999)
Esterase-assisted accumulation of 3-carboxy-2,2,5,5-tetramethyl-1-pyrrolidinyloxyl into lymphocytes.
J. Kao (2004)
The role of cytosolic free calcium, (Ca2+)i, and phosphoinositides as intracellular signals
P. Lew (1987)
Calcium Signaling Protocols
J. Walker (2005)
Methodological aspects of pressure loading of Fura-2 into Characean cells
C. Plieth (1996)
Effects of calmodulin antagonists on taurine-stimulated calcium ion uptake in the rat retina are partly independent of calmodulin activity.
Julius D. Militante (2000)
Two-Photon Microscopy with Diffractive Optical Elements and Spatial Light Modulators
Brendon O. Watson (2010)
Effects of calcium chelators on intracellular calcium and excitotoxicity
J. Dubinsky (1993)
Measurement of intracellular calcium levels of human spermatozoa acrosome reacted by electropermeabilization.
M. Wall (1994)
Cellular and subcellular calcium signaling in gastrointestinal epithelium.
M. Nathanson (1994)
Astroglial excitability and gliotransmission: an appraisal of Ca2+ as a signalling route
R. Zorec (2012)
5 – Cellular Calcium and Phospholipid Metabolism Mediate Pancreatic Enzyme Secretion
S. Pandol (1990)
The possible pitfalls of the measurement of intracellular calcium concentration of lymphocytes with the fluorescent indicator quin2.
P. Csermely (1987)
Chapter 6 Talking to cells—cell membrane receptors and their modes of action
R. Irvine (1997)
alpha1-adrenergic stimulation induces early afterdepolarizations in ferret Purkinje fibers.
E. Drouin (1996)
Physiological implications of the interaction between the plasma membrane calcium pump and nNOS
E. Cartwright (2008)
Presynaptic strontium dynamics and synaptic transmission.
M. A. Xu-Friedman (1999)
Screening Strategies for the Discovery of Ion Channel Monoclonal Antibodies
Caroline S Colley (2018)
Physiology and pathophysiology of the calcium store in the endoplasmic reticulum of neurons.
A. Verkhratsky (2005)
Measurement of intracellular ionic composition and activities in renal tubules.
W. Boron (1983)
Histone deacetylase 6 inhibition reduces cysts by decreasing cAMP and Ca2+ in knock-out mouse models of polycystic kidney disease
Murali K Yanda (2017)
Inhibition of Mitochondrial Bioenergetics by Esterase-Triggered COS/H2S Donors.
A. Steiger (2017)See more