Ins(1,4,5)P3 activates Drosophila cation channel Trpl in recombinant baculovirus-infected Sf9 insect cells.
The trp-like (trpl) gene product (Trpl) is thought to form a nonselective cation channel important for signal transduction in Drosophila photoreceptor cells. This channel may be the insect homologue of mammalian channels involved in Ca2+ signal transduction. To determine the mechanism of receptor-mediated activation of Trpl, whole cell membrane currents were examined in Sf9 insect cells after infection with recombinant baculovirus. Stimulation by bradykinin increased whole cell Trpl currents three- to fivefold. Similar activation of Trpl was observed by inclusion of D-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] in the pipette solution during whole cell recordings. These currents were 1) not seen in noninfected cells or in cells expressing only the B2 receptor, 2) mimicked by D-myo-inositol 2,4,5-trisphosphate, and 3-deoxy-3-fluoro-D-myo-inositol 1,4,5-trisphosphate, 3) not seen with D-myo-inositol 1,4-bisphosphate or D-myo-inositol 1,3,4,5-tetrakisphosphate, and 4) blocked by heparin, but not by de-N-sulfated heparin. In contrast, Trpl currents were unaffected by thapsigargin. These results demonstrate that the Trpl cation channel is activated by Ins(1,4,5)P3 in a heparin-sensitive fashion. Regulation of channel activity by Ins(1,4,5)P3 may occur by a number of mechanisms, including direct binding of Ins(1,4,5)P3 to the Trpl channel or direct physical interaction between the Ins(1,4,5)P3 receptor/Ca(2+)-release channel of the endoplasmic reticulum and the Trpl protein.
D000818:Animals; D016367:Baculoviridae; D002148:Calmodulin-Binding Proteins / Q000378:metabolism* / Q000502:physiology; D002412:Cations / Q000378:metabolism*; D002460:Cell Line; D004330:Drosophila / Q000378:metabolism*; D029721:Drosophila Proteins; D004553:Electric Conductivity; D015544:Inositol 1,4,5-Trisphosphate / Q000494:pharmacology*; D007313:Insecta; D007473:Ion Channels / Q000187:drug effects*; D008565:Membrane Proteins / Q000378:metabolism* / Q000502:physiology; D011956:Receptors, Cell Surface / Q000502:physiology; D011995:Recombination, Genetic; D050051:Transient Receptor Potential Channels; D014777:Virus Diseases / Q000378:metabolism / Q000473:pathology
D002148:Calmodulin-Binding Proteins; D002412:Cations; D029721:Drosophila Proteins; D007473:Ion Channels; D008565:Membrane Proteins; D011956:Receptors, Cell Surface; D050051:Transient Receptor Potential Channels; C074192:trpl protein, Drosophila; D015544:Inositol 1,4,5-Trisphosphate