Inositol tetrakisphosphate liberates stored Ca2+ in Xenopus oocytes and facilitates responses to inositol trisphosphate.

1. The actions of the putative second messenger inositol 1,3,4,5-tetrakisphosphate (Ins(1,3,4,5)P4) were studied by injecting it into voltage-clamped oocytes while recording Ca(2+)-dependent chloride membrane currents and, in some experiments, fluorescence signals from Ca2+ indicators. 2. Ins(1,3,4,5)P4 evoked a rise in intracellular Ca2+ and associated chloride current in oocytes bathed in normal or Ca(2+)-free Ringer solutions. The fluorescence Ca2+ signal showed a prolonged rise with superimposed oscillations, whereas the current reflected only the oscillatory component. 3. Injections of inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) evoked currents showing an initial transient, followed by oscillations. Ins(1,3,4,5)P4 evoked similar oscillations, but the transient component was usually small or absent. Ins(1,3,4,5)P4 was about 20-fold less potent than Ins(1,4,5)P3, as measured by comparing doses required to elicit currents with the same integral. The most sensitive oocytes responded to about 1 fmol Ins(1,3,4,5)P4 and 0.1 fmol Ins(1,4,5)P3. 4. Injections of Ins(2,4,5)P4 evoked oscillatory currents, with a potency about three times greater than Ins(1,4,5)P3. Ins(1,3,4)P4 was ineffective in some oocytes even at doses of several picomoles, but in other oocytes evoked small transient and oscillatory currents with a potency 100 times or more less than Ins(1,3,4,5)P4. 5. Injections of Ins(1,3,4,5)P4 made into the animal hemisphere of the oocyte evoked larger currents than injections into the vegetal hemisphere. 6. Photo-release of Ins(1,4,5)P3 from caged Ins(1,4,5)P4 loaded into the oocyte was used to examine interactions between Ins(1,4,5)P3 and Ins(1,3,4,5)P4. Injection of low (ca 1 fmol) doses of Ins(1,3,4,5)P4 shortly before a light flash greatly facilitated currents evoked by photo-release of near-threshold amounts of Ins(1,4,5)P3. This facilitation was unaffected by removal of extracellular Ca2+ and arose because Ins(1,3,4,5)P4 reduced the threshold amount of Ins(1,4,5)P3 required to evoke a response. 7. Larger amounts (several femtomoles) of Ins(1,3,4,5)P4 depressed responses evoked by photo-release of Ins(1,4,5)P3. This may arise because Ca2+ liberated by Ins(1,3,4,5)P4 inhibits the ability of Ins(1,4,5)P3 to release further Ca2+. 8. We conclude that Ins(1,3,4,5)P4 liberates intracellular Ca2+ in the oocyte in a manner similar to that of Ins(1,4,5)P3, and suggest that a physiological role for Ins(1,3,4,5)P4 may be to facilitate responses to Ins(1,4,5)P3.