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© The Rockefeller University Press, 0021-9525/1997//93 $5.00
The Journal of Cell Biology, Volume 137, Number 1, , 1997 93-104

Article

Lysosomes Behave as Ca2+-regulated Exocytic Vesicles in Fibroblasts and Epithelial Cells



Ana Rodríguez*, Paul Webster*, Javier Ortego{ddagger}, and Norma W. Andrews*

* Department of Cell Biology, {ddagger} Department of Ophthalmology, Yale University School of Medicine, New Haven, Connecticut 06520

Lysosomes are considered to be a terminal degradative compartment of the endocytic pathway, into which transport is mostly unidirectional. However, specialized secretory vesicles regulated by Ca2+, such as neutrophil azurophil granules, mast cell–specific granules, and cytotoxic lymphocyte lytic granules, share characteristics with lysosomes that may reflect a common biogenesis. In addition, the involvement of Ca2+ transients in the invasion mechanism of the parasite Trypanosoma cruzi, which occurs by fusion of lysosomes with the plasma membrane, suggested that lysosome exocytosis might be a generalized process present in most cell types.

Here we demonstrate that elevation in the intracellular free Ca2+ concentration of normal rat kidney (NRK) fibroblasts induces fusion of lysosomes with the plasma membrane. This was verified by measuring the release of the lysosomal enzyme β-hexosaminidase, the appearance on the plasma membrane of the lysosomal glycoprotein lgp120, the release of fluid-phase tracers previously loaded into lysosomes, and the release of the lysosomally processed form of cathepsin D. Exposure to the Ca2+ ionophore ionomycin or addition of Ca2+containing buffers to streptolysin O–permeabilized cells induced exocytosis of ~10% of the total lysosomes of NRK cells. The process was also detected in other cell types such as epithelial cells and myoblasts. Lysosomal exocytosis was found to require micromolar levels of Ca2+ and to be temperature and ATP dependent, similar to Ca2+-regulated secretory mechanisms in specialized cells.

These findings highlight a novel role for lysosomes in cellular membrane traffic and suggest that fusion of lysosomes with the plasma membrane may be an ubiquitous form of Ca2+-regulated exocytosis.

Abbreviations used in this paper: [Ca2+]i, intracellular free Ca2+ concentration; LDH, lactate dehydrogenase; lgp, lysosomal membrane glycoprotein; NRK, normal rat kidney; SLO, streptolysin O; TSF, trypanosomesoluble fraction.

Please address all correspondence to Norma W. Andrews, Department of Cell Biology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520. Tel.: (203) 785-4314. Fax: (203) 785-7226.


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