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Ca2+ Dynamics in the Secretory Vesicles of Neurosecretory PC12 and INS1 Cells

AuthorsSanto-Domingo, Jaime; Fonteriz, Rosalba I. ; Lobatón, Carmen D.; Montero, Mayte ; Moreno, Alfredo; Álvarez, Javier
Secretory granules
PC12 cells
INS1 cells
Ryanodine receptors
Issue DateNov-2010
CitationCellular and Molecular Neurobiology 30(8): 1267-1274 (2010)
AbstractWe have investigated the dynamics of the free [Ca2+] inside the secretory granules of neurosecretory PC12 and INS1 cells using a low-Ca2+-affinity aequorin chimera fused to synaptobrevin-2. The steady-state secretory granule [Ca2+] ([Ca2+]SG] was around 20–40 μM in both cell types, about half the values previously found in chromaffin cells. Inhibition of SERCA-type Ca2+ pumps with thapsigargin largely blocked Ca2+ uptake by the granules in Ca2+-depleted permeabilized cells, and the same effect was obtained when the perfusion medium lacked ATP. Consistently, the SERCA-type Ca2+ pump inhibitor benzohydroquinone induced a rapid release of Ca2+ from the granules both in intact and permeabilized cells, suggesting that the continuous activity of SERCA-type Ca2+ pumps is essential to maintain the steady-state [Ca2+]SG. Both inositol 1,4,5-trisphosphate (InsP3) and caffeine produced a rapid Ca2+ release from the granules, suggesting the presence of InsP3 and ryanodine receptors in the granules. The response to high-K+ depolarization was different in both cell types, a decrease in [Ca2+]SG in PC12 cells and an increase in [Ca2+]SG in INS1 cells. The difference may rely on the heterogeneous response of different vesicle populations in each cell type. Finally, increasing the glucose concentration triggered a decrease in [Ca2+]SG in INS1 cells. In conclusion, our data show that the secretory granules of PC12 and INS1 cells take up Ca2+ through SERCA-type Ca2+ pumps and can release it through InsP3 and ryanodine receptors, supporting the hypothesis that secretory granule Ca2+ may be released during cell stimulation and contribute to secretion.
Description8 páginas, 4 figuras.-- Trabajo presentado al: "15th International Symposium on Chromaffin Cell Biology: The Chromaffin Cell as a Stress Transducer".-- el pdf del artículo es la versión manuscrita de autor.
Publisher version (URL)http://dx.doi.org/10.1007/s10571-010-9572-2
Appears in Collections:(IBGM) Artículos
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