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J Biol Chem, Vol. 273, Issue 44, 28657-28662, October 30, 1998
From the Department of Physiology and Biophysics University of
California, Irvine, California 92697-4560
To measure the concentration of inositol
1,4,5-trisphosphate ([IP3]) in small regions of
single Xenopus oocytes, a biological detector cell was
combined with capillary electrophoresis. This method is 10,000 times
more sensitive than all existing assays enabling subcellular
measurement of [IP3] in Xenopus oocytes. Upon
addition of lysophosphatidic acid to an oocyte, [IP3]
increased from 40 to 650 nM within 2 min. IP3
concentrations as high as 1.8 µM were measured after
activation with lysophosphatidic acid, suggesting that the physiologic
concentration of IP3 ranges from the tens of nanomolar to a
few micromolar in Xenopus oocytes. Since the
IP3 receptor in Xenopus oocytes is nearly
identical to the type I receptor of mammalian cells, the range of
[IP3] in most mammalian cells is likely to be similar to
that in the oocyte. By selecting or engineering the appropriate
detector cell, this strategy should be applicable to cyclic adenosine
diphosphate ribose and nicotinic acid adenine dinucleotide phosphate,
and to the discovery of new Ca2+-releasing second
messengers.
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