Subcellular distribution of the calcium-storing inositol 1,4,5-trisphosphate-sensitive organelle in rat liver. Possible linkage to the plasma membrane through the actin microfilaments.
The role of Ins(1,4,5)P3 in the mobilization of Ca2+ from intracellular stores of non-muscle cells has been extensively demonstrated; however, the nature of the organelle releasing the Ca2+ is still poorly understood. The distributions of the Ins(1,4,5)P3-binding sites and of the Ins(1,4,5)P3-sensitive Ca2+ pool were investigated in subcellular fractions obtained from rat liver and compared with those of other markers. The Ins(1,4,5)P3-binding vesicles appeared to be completely distinct from the endoplasmic-reticulum-derived microsomes and were enriched in the same fractions which were enriched in alkaline phosphodiesterase I activity. This co-purification of the plasma-membrane marker with the Ins(1,4,5)P3-binding sites was dramatically altered after freezing or after treatment of the homogenate with the microfilament-disruptive drug cytochalasin B, suggesting that the Ins(1,4,5)P3-sensitive organelle may be linked to the plasma membrane through the actin microfilaments. No correlation was observed between the Ins(1,4,5)P3-binding capacity and the portion of the Ca2+ pool that was released by Ins(1,4,5)P3. This may result from the disruption of the native organelle during homogenization, leading to the formation of vesicles containing the Ins(1,4,5)P3 receptor, but lacking the Ca2+ pump. These results are consistent with the idea of a specialized Ins(1,4,5)P3-regulated organelle distinct from the endoplasmic reticulum, and we propose a model of the structural organization of this organelle, in which the anchorage to the cytoskeleton as well as the spatial separation of the Ca2+ pump from the Ins(1,4,5)P3 receptor have important functional significance.
Rossier
M F
MF
Laboratory of Cellular and Molecular Pharmacology, NIEHS/NIH, Research Triangle Park, NC 27709.
Bird
G S
GS
Putney
J W
JW
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels; D002458:Cell Fractionation; D003571:Cytochalasin B / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism*; D053496:Inositol 1,4,5-Trisphosphate Receptors; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D015388:Organelles / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D011956:Receptors, Cell Surface / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear
D015220:Calcium Channels; D053496:Inositol 1,4,5-Trisphosphate Receptors; D011956:Receptors, Cell Surface; D018160:Receptors, Cytoplasmic and Nuclear; D003571:Cytochalasin B; D015544:Inositol 1,4,5-Trisphosphate; D002118:Calcium