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. 1996 Nov 1;319(Pt 3):909–912. doi: 10.1042/bj3190909

The lysosomal Ca2+ pool in MDCK cells can be released by ins(1,4,5)P3-dependent hormones or thapsigargin but does not activate store-operated Ca2+ entry.

T Haller 1, H Völkl 1, P Deetjen 1, P Dietl 1
PMCID: PMC1217874  PMID: 8920998

Abstract

In several cell types, Ca2+ release from intracellular Ca2+ stores by Ins(1,4,5)P3 elicits Ca2+ influx from the extracellular space into the cytoplasm, termed store-operated Ca2+ entry (SOCE). In MDCK cells, the Ins(1,4,5)P3-sensitive Ca2+ store giving rise to SOCE essentially overlaps with the thapsigargin (TG)-sensitive store. Recent evidence suggests that in MDCK cells lysosomes form a Ca2+ pool that is functionally coupled with the Ins(1,4,5)P3-sensitive Ca2+ store: Ca2+ can be selectively released from lysosomes by glycyl-L-phenylalanine naphthylamide, an agent inducing lysosomal swelling with subsequent and reversible permeabilization of the vesicular membranes. This compartment is also depleted by Ins(1,4,5)P3-dependent agonists or TG, indicating that it is part of a larger, Ins(1,4,5)P3-sensitive Ca2+ pool. Here we show that whereas SOCE is triggered by Ca2+ release from the entire Ins(1,4,5)P3-sensitive Ca2+ pool, selective Ca2+ release from lysosomes alone is unable to trigger SOCE. This finding is consistent with measurements of the store-operated cation current, a direct parameter for store-operated Ca2+ and Na+ entry into MDCK cells. Hence it is proposed that the Ins(1,4,5)P3-sensitive Ca2+ pool is composed of different intracellular compartments that do not uniformly stimulate Ca2+ entry into the cell.

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Selected References

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  1. Berg T. O., Strømhaug E., Løvdal T., Seglen O., Berg T. Use of glycyl-L-phenylalanine 2-naphthylamide, a lysosome-disrupting cathepsin C substrate, to distinguish between lysosomes and prelysosomal endocytic vacuoles. Biochem J. 1994 May 15;300(Pt 1):229–236. doi: 10.1042/bj3000229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berg T. O., Strømhaug P. E., Berg T., Seglen P. O. Separation of lysosomes and autophagosomes by means of glycyl-phenylalanine-naphthylamide, a lysosome-disrupting cathepsin-C substrate. Eur J Biochem. 1994 Apr 1;221(1):595–602. doi: 10.1111/j.1432-1033.1994.tb18771.x. [DOI] [PubMed] [Google Scholar]
  3. Berridge M. J. Capacitative calcium entry. Biochem J. 1995 Nov 15;312(Pt 1):1–11. doi: 10.1042/bj3120001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Delles C., Haller T., Dietl P. A highly calcium-selective cation current activated by intracellular calcium release in MDCK cells. J Physiol. 1995 Aug 1;486(Pt 3):557–569. doi: 10.1113/jphysiol.1995.sp020834. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fasolato C., Innocenti B., Pozzan T. Receptor-activated Ca2+ influx: how many mechanisms for how many channels? Trends Pharmacol Sci. 1994 Mar;15(3):77–83. doi: 10.1016/0165-6147(94)90282-8. [DOI] [PubMed] [Google Scholar]
  6. Ghosh T. K., Mullaney J. M., Tarazi F. I., Gill D. L. GTP-activated communication between distinct inositol 1,4,5-trisphosphate-sensitive and -insensitive calcium pools. Nature. 1989 Jul 20;340(6230):236–239. doi: 10.1038/340236a0. [DOI] [PubMed] [Google Scholar]
  7. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  8. Hajnóczky G., Lin C., Thomas A. P. Luminal communication between intracellular calcium stores modulated by GTP and the cytoskeleton. J Biol Chem. 1994 Apr 8;269(14):10280–10287. [PubMed] [Google Scholar]
  9. Haller T., Dietl P., Deetjen P., Völkl H. The lysosomal compartment as intracellular calcium store in MDCK cells: a possible involvement in InsP3-mediated Ca2+ release. Cell Calcium. 1996 Feb;19(2):157–165. doi: 10.1016/s0143-4160(96)90084-6. [DOI] [PubMed] [Google Scholar]
  10. Hampe W., Zimmermann P., Schulz I. GTP-induced fusion of isolated pancreatic microsomal vesicles is increased by acidification of the vesicle lumen. FEBS Lett. 1990 Oct 1;271(1-2):62–66. doi: 10.1016/0014-5793(90)80372-p. [DOI] [PubMed] [Google Scholar]
  11. Jacob R. Agonist-stimulated divalent cation entry into single cultured human umbilical vein endothelial cells. J Physiol. 1990 Feb;421:55–77. doi: 10.1113/jphysiol.1990.sp017933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jadot M., Colmant C., Wattiaux-De Coninck S., Wattiaux R. Intralysosomal hydrolysis of glycyl-L-phenylalanine 2-naphthylamide. Biochem J. 1984 May 1;219(3):965–970. doi: 10.1042/bj2190965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jadot M., Wattiaux R. Effect of glycyl-L-phenylalanine 2-naphthylamide on invertase endocytosed by rat liver. Biochem J. 1985 Feb 1;225(3):645–648. doi: 10.1042/bj2250645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Penner R., Fasolato C., Hoth M. Calcium influx and its control by calcium release. Curr Opin Neurobiol. 1993 Jun;3(3):368–374. doi: 10.1016/0959-4388(93)90130-q. [DOI] [PubMed] [Google Scholar]
  15. Pozzan T., Rizzuto R., Volpe P., Meldolesi J. Molecular and cellular physiology of intracellular calcium stores. Physiol Rev. 1994 Jul;74(3):595–636. doi: 10.1152/physrev.1994.74.3.595. [DOI] [PubMed] [Google Scholar]
  16. Putney J. W., Jr, Bird G. S. The signal for capacitative calcium entry. Cell. 1993 Oct 22;75(2):199–201. doi: 10.1016/0092-8674(93)80061-i. [DOI] [PubMed] [Google Scholar]
  17. Rossier M. F., Bird G. S., Putney J. W., Jr 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. Biochem J. 1991 Mar 15;274(Pt 3):643–650. doi: 10.1042/bj2740643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Volpe P., Krause K. H., Hashimoto S., Zorzato F., Pozzan T., Meldolesi J., Lew D. P. "Calciosome," a cytoplasmic organelle: the inositol 1,4,5-trisphosphate-sensitive Ca2+ store of nonmuscle cells? Proc Natl Acad Sci U S A. 1988 Feb;85(4):1091–1095. doi: 10.1073/pnas.85.4.1091. [DOI] [PMC free article] [PubMed] [Google Scholar]

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