Abstract
Electrophysiological studies in some secretory and non-secretory cells have identified an extensive form of calcium-induced exocytosis that is rapid (hundreds of milliseconds), insensitive to tetanus toxin and distinct from regulated secretion. We have now identified a marker of the process, desmoyokin-AHNAK, in a clonal derivative of the neuronal cell line, PC12. In resting cells, desmoyokin-AHNAK is localized within the lumen of specific vesicles, but appears on the cell surface during stimulation. Desmoyokin-AHNAK-positive vesicles exist in a variety of cells and tissues and are distinct from the endoplasmic reticulum, Golgi, trans-Golgi, endosomes and lysosomes, and from Glut4 and constitutive secretion vesicles. They seem to be involved in two models of plasmalemma enlargement: differentiation and membrane repair. We therefore propose that these vesicles should be called 'enlargosomes'.
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Acknowledgements
We thank H. Kasai and his laboratory staff for their pioneering collaboration and advice; E. Shtivelman and M. Rupnik for generous support; M. Matteoli and G.P. Schiavo for advice, A. Lorusso for participating in some experiments; C. Paolucci for FACS analysis, R. Barsacchi for performing the [Ca2+]i measurements; and D. Dunlap for the critical revision of the text. Imaging experiments were performed within Alembic (Advanced Light and Electron Microscopy Bio-Imaging Center), San Raffaele Scientific Institute. This work, supported by grants from Telethon, the European Union (QLG1–02233), CNR (Ag. 2000 and the CNS degenerative diseases), the Cofin Italian University System (2001.053389-033) and the Armenise–Harvard Foundation, was performed in the framework of the Italian MIUR Center of Excellence in Physiopathology of Cell Differentiation.
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Supplementary figures
SI Figure 1 Distinct labeling patterns of PC12 cells with the anti-dA FEN and KIS antibodies. blotting (D and E). (PDF 1678 kb)
SI Figure 2 Surface appearance of dA in stimulated living cells.
SI Figure 3 Cell types investigated for dA expression
Movie I
The movie illustrates in 3D rendering the distribution of the punctate dA surface staining in a PC12-27 cell first stimulated by caged Ca2+ photolysis and then immunolabeled while still living. (GIF 315 kb)
Movie 2
The movie illustrates in 3D rendering the distribution of neurosecretory and "enlargesomal" vesicles in a wild- type PC12 cell differentiated with NGF. (AVI 2820 kb)
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Borgonovo, B., Cocucci, E., Racchetti, G. et al. Regulated exocytosis: a novel, widely expressed system. Nat Cell Biol 4, 955–963 (2002). https://doi.org/10.1038/ncb888
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DOI: https://doi.org/10.1038/ncb888
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