Cell Calcium 1994 Jul 01;161:20-8. doi: 10.1016/s0143-4160(05)80004-1.

Independent external calcium entry and cellular calcium mobilization in Xenopus oocytes.

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We studied cellular calcium (Ca) mobilization and Ca entry from the medium following injection of various inositol phosphates (IPs) or activation of thyrotropin-releasing hormone receptors (TRH-Rs) in oocytes injected with TRH-R cRNA. We determined the order of potency of various IPs for evoking the rapid depolarizing current in Ca-free medium, which reflects the mobilization of cellular Ca. The most potent compound was inositol 1,4,5-trisphosphate (Ins(1,4,5)P3), followed by inositol 1,2,4,5-tetrakisphosphate (Ins(1,2,4,5)-P4), which displayed 91% of the activity of Ins(1,4,5)P3, while inositol 1,3,4,6-tetrakisphosphate (Ins(1,3,4,6)P4) had only 29% effect. All other IPs used in the present study exhibited responses that were 40% or less than those elicited by Ins(1,4,5)P3. Cellular Ca mobilization was confirmed by 45Ca2+ efflux for Ins(1,4,5)P3, Ins(1,2,4,5)P4 and Ins(1,3,4,6)P4, or by Fura-2 ratio imaging studies for the latter. In parallel, we assayed the ability of these compounds to promote Ca entry into the cell, as reflected by Ca-evoked depolarizing current or Fura-2 imaging. These assays revealed a different order of potency, where Ins(1,4,5)P3 > inositol 4,5-bisphosphate (Ins(4,5)P2) > Ins(1,3,4,6)P4 = Ins(1,2,4,5)P4. All other inositol phosphates were largely ineffective. Heparin inhibited the response to TRH by 67% while Ca entry was inhibited only by 22%. The latency of the response to TRH was significantly shorter in the presence of extracellular Ca, suggesting Ca entry preceded the response, i.e. major depletion of Ca stores. These results strongly suggest that the activation of Ca entry is largely independent of cellular Ca mobilization and may be mediated by a receptor for an unidentified phosphorylated compound, different from that for Ins(1,4,5)P3 on the endoplasmic reticulum.

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Species referenced: Xenopus
Genes referenced: isyna1 trh