Premium
The measurement of intracellular calcium levels in protoplasts from higher plant cells
Author(s) -
Gilroy S.,
Hughes W.A.,
Trewavas A.J.
Publication year - 1986
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(86)80483-5
Subject(s) - protoplast , egta , electroporation , calcium , plant cell , intracellular , incubation , biophysics , cytosol , membrane potential , membrane , chemistry , biochemistry , biology , enzyme , organic chemistry , gene
The measurement of cytosolic calcium concentration, [Ca 2+ ] i , in the higher plant cells has proved difficult due to the negligible uptake of [Ca 2+ ] i indicator. The uptake of the fluorescent [Ca 2+ ] i indicator, quin 2, as its permeant ester, quin 2/AM, proved unsuccessful when used with plant cells and cell protoplasts. However, electrically induced membrane permeabilisation, electroporation, has allowed quin 2 uptake into mung bean root protoplasts to 10 −4 M. Resting [Ca 2+ ] was measured as 171 ± 41 nM. The loaded quin 2 was responsive to changes in the Ca 2+ environment of the protoplasts, indicating a reversible fall in [Ca 2+ ] to 17% of the initial value, over 10 min, on incubation with EGTA.