The measurement of intracellular calcium levels in protoplasts from higher plant cells

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 cel...

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Bibliographic Details
Published in:FEBS letters 1986-04, Vol.199 (2), p.217-221
Main Authors: Gilroy, S., Hughes, W.A., Trewavas, A.J.
Format: Article
Language:English
Subjects:
Applied sciences
Calcium estimation
DMSO, dimethyl sulphoxide
DTPA, diethylenetriaminepentaacetic acid
Electroporation
Exact sciences and technology
FDA, fluorescein diacetate
Mung bean root
Other techniques and industries
Protoplast
Quin 2
quin 2, methoxyquinolinebis-(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid
quin 2/AM, tetraacetoxymethyl ester of quin 2
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Summary: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.
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(86)80483-5