PMID: 2117051Sep 1, 1990Paper

Use of 1,2-bis(2-amino-5-fluorophenoxy)ethane-N,N,N',N'-tetraacetic acid (5FBAPTA) in the measurement of free intracellular calcium in the brain by 19F-nuclear magnetic resonance spectroscopy

Journal of Neurochemistry
R S Badar-GofferH S Bachelard

Abstract

We have applied the 19F-nuclear magnetic resonance (NMR) calcium indicator 1,2-bis(2-amino-5-fluoro-phenoxy)ethane-N,N,N',N'-tetraacetic acid (5FBAPTA) to the measurement of the free intracellular calcium concentration [( Ca2+]i) in superfused brain slices. A mean +/- SD control value of 380 +/- 71 nM (n = 18) was obtained at 37 degrees C using 2.4 mM extracellular Ca2+. Subcellular fractionation studies using [3H]5FBAPTA showed that after loading of its tetraacetoxymethyl ester, approximately 55% was de-esterified, with the other 45% remaining as the tetraester bound to membranes. Of the de-esterified 5FBAPTA, greater than 90% was in the cytosolic fractions, with less than 1% in the mitochondria or microsomes. The NMR-visible de-esterified 5FBAPTA slowly disappeared from the tissue with a t1/2 of 4 h. A time course after loading confirmed that the calculated [Ca2+]i was constant over a 5-h period, although the scatter of individual results was +/- 20%. The [Ca2+]i was increased by a high extracellular K+ concentration ([K+]e), by a low extracellular concentration of Na+, and by the calcium ionophore A23187. On recovery from high [K+]e, the [Ca2+]i "overshot" to values lower than the original control value. The [Ca2+]i was surp...Continue Reading

References

Jan 29, 1986·Biochimica Et Biophysica Acta·R H Ashley
Dec 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·H L KirschenlohrG A Smith
Aug 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·J A Connor
Oct 1, 1985·Biochemical Society Transactions·H S BachelardP G Morris
Apr 1, 1974·Journal of Neurochemistry·G M Heaton, H S Bachelard
Jan 1, 1983·Annual Review of Physiology·R DiPolo, L Beaugé
Apr 1, 1984·The Biochemical Journal·R H AshleyR Marchbanks
Apr 16, 1984·Biochimica Et Biophysica Acta·C D RichardsT R Hesketh

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Citations

Dec 1, 1993·Biophysical Journal·R K Gupta, B A Wittenberg
Aug 1, 1993·Journal of Neurochemistry·H Bachelard, R Badar-Goffer
Jun 1, 1999·Journal of Neurochemistry·S A AndersonR S Hotchkiss
Aug 7, 2018·Chemical Reviews·Zhenchuang XuYanchuan Zhao

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