Abstract
There are only a few techniques for measuring cytoplasmic free calcium and in general these are technically difficult and may only have limited applicability. In the past few years a new generation of fluorescent indicators have been developed (Tsien 1980) which promise to be of great value. The first of these indicators to be commercially available is quin-2. While there are some inherent disadvantages in the detection of quin-2 in cells, the main advantage of the method lies in the use of the membrane- permeable acetoxymethyl ester (quin-2-AM) which should be hydrolysed by cytoplasmic esterases to trap the fluorescent indicator in the cell (Tsien 1981). This method of loading cells is much less disruptive than microinjection and should be applicable to a whole range of cells which cannot be microinjected or have microelectrodes inserted in them. It has been used on a variety of animal cells but as yet there are no published reports of its use in plant cells. I describe here some attempts to use quin-2-AM on a number of plant cells, outline some of the problems found and suggest some possible answers.
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References
Hesketh, T.R., Smith, G.A., Moore, J.P., Taylor, M.V. and Metcalfe, J.C., 1983, Free cytoplasmic calcium concentration and the mitogenic stimulation of lymphocytes, J. Biol. Chem., 258: 4876.
Rogers, J., Hesketh, T.R., Smith, G.A., Beaven, M.A., Metcalfe, J.C.,Johnson, P. and Garland, P.B., 1983, Intracellular pH and free calcium changes in single cells using quene 1 and quin 2 probes and fluores¬cence microscopy. FEBS Letts, 161: 21.
Tsien, R.Y., 1980, New calcium indicators and buffers with high selectivity against magnesium and protons. Design, synthesis and properties of prototype structures, Biochemistry, 19: 2396.
Tsien, R.Y., 1981, A non-disruptive technique for loading calcium buffer nd indicators into cells, Nature, 290: 527.
Tsien, R.Y., Pozzan, T. and Rink, T.J., 1982, Calcium homeostasis in intact lymphocytes: cytoplasmic free calcium measured with a new, intracellular trapped fluorescent indication, J. Cell Biol., 94: 325.
Tsien, R.Y., Pozzan, T. and Rink, T.J., 1984, Measuring and manipulating cytosolic Ca2+ with trapped indicators, TIBS, 9 (6): 263.
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© 1986 Plenum Press, New York
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Cork, R.J. (1986). An Assessment of the Usefulness of Quin-2-AM for Cytoplasmic Calcium Measurements in Plant Cells. In: Trewavas, A.J. (eds) Molecular and Cellular Aspects of Calcium in Plant Development. NATO ASI Series, vol 104. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-2177-4_65
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DOI: https://doi.org/10.1007/978-1-4613-2177-4_65
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