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J Biol Chem, Vol. 273, Issue 44, 28657-28662, October 30, 1998

The Physiologic Concentration of Inositol 1,4,5-Trisphosphate in the Oocytes of Xenopus laevis

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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