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Abstract:

Neurons and neuroendocrine cells must retrieve plasma membrane excess and refill vesicle pools depleted by exocytosis. To perform these tasks cells can use different endocytosis/recycling mechanisms whose selection will impact on vesicle recycling time and secretion performance. We used FM1-43 to evaluate in the same experiment exocytosis, endocytosis, and recovery of releasable vesicles on mouse chromaffin cells. Various exocytosis levels were induced by a variety of stimuli, and we discriminated the resultant endocytosis-recycling responses according to their ability to rapidly generate releasable vesicles. Exocytosis of ≤20% of plasma membrane (provoked by nicotine/acetylcholine) was followed by total recovery of releasable vesicles. If a stronger stimulus (50 mM K + and 2 mM Ca2+) provoking intense exocytosis (51 ± 7%) was applied, endocytosis still retrieved all the fused membrane, but only a fraction (19 ± 2%) was releasable by a second stimulus. Using ADVASEP-7 or bromophenol blue to quickly eliminate fluorescence from noninternalized FM1-43, we determined that this fraction became releasable in <2 min. The remaining nonreleasable fraction was distributed mainly as fluorescent spots (∼0.7 μm) selectively labeled by 40- to 70-kDa dextrans and was suppressed by a phosphatidylinositol-3-phosphate kinase inhibitor, suggesting that it had been formed by a bulk retrieval mechanism. We concluded that chromaffin cells can rapidly recycle significant fractions of their total vesicle population, and that this pathway prevails when cholinergic agonists are used as secretagogues. When exocytosis exceeded ∼20% of plasma membrane, an additional mechanism was activated, which was unable to produce secretory vesicles in our experimental time frame but appeared crucial to maintaining membrane surface homeostasis under extreme conditions. Copyright © 2007 the American Physiological Society.

Registro:

Documento: Artículo
Título:Rapid recovery of releasable vesicles and formation of nonreleasable endosomes follow intense exocytosis in chromaffin cells
Autor:Bay, A.E.P.; Ibañez, L.I.; Marengo, F.D.
Filiación:Laboratorio de Fisiología Y Biología Molecular, Inst. de Fisiol., Biologia Molec. Y Neurociencias (Consejo Nac. de Invest. Cientificas Y Tecnicas), Universidad de Buenos Aires, Buenos Aires, Argentina
Laboratorio de Fisiología Y Biología Molecular, Instituto de Fisiología, Biología Molecular Y Neurociencias, Ciudad Universitaria-Pabellón II, CP 1428, Buenos Aires, Argentina
Palabras clave:ADVASEP-7; Bromophenol blue; Calcium signal; Endocytosis; FM1-43; Mouse chromaffin cells; 2 morpholino 8 phenylchromone; acetylcholine; calcium ion; fluorescent dye; FMI 43; nicotine; phosphatidylinositol 3 kinase inhibitor; unclassified drug; adrenal gland; animal cell; article; chromaffin cell; controlled study; endocytosis; endosome; exocytosis; fluorescence; homeostasis; membrane vesicle; mouse; neurosecretory cell; nonhuman; priority journal; 1-Phosphatidylinositol 3-Kinase; Acetylcholine; Adrenal Glands; Animals; Bromphenol Blue; Calcium; Cells, Cultured; Cholinergic Agonists; Chromaffin Cells; Cyclodextrins; Endocytosis; Endosomes; Exocytosis; Fluorescent Dyes; Homeostasis; Membrane Fusion; Mice; Nicotine; Potassium; Protein Kinase Inhibitors; Pyridinium Compounds; Quaternary Ammonium Compounds; Staining and Labeling; Time Factors; Transport Vesicles
Año:2007
Volumen:293
Número:5
Página de inicio:C1509
Página de fin:C1522
DOI: http://dx.doi.org/10.1152/ajpcell.00632.2006
Título revista:American Journal of Physiology - Cell Physiology
Título revista abreviado:Am. J. Physiol. Cell Physiol.
ISSN:03636143
CODEN:AJPCD
CAS:2 morpholino 8 phenylchromone, 154447-36-6; acetylcholine, 51-84-3, 60-31-1, 66-23-9; calcium ion, 14127-61-8; nicotine, 54-11-5; 1-Phosphatidylinositol 3-Kinase, 2.7.1.137; ADVASEP 7; Acetylcholine, 51-84-3; Bromphenol Blue, 115-39-9; Calcium, 7440-70-2; Cholinergic Agonists; Cyclodextrins; FM1 43; Fluorescent Dyes; Nicotine, 54-11-5; Potassium, 7440-09-7; Protein Kinase Inhibitors; Pyridinium Compounds; Quaternary Ammonium Compounds
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03636143_v293_n5_pC1509_Bay

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Citas:

---------- APA ----------
Bay, A.E.P., Ibañez, L.I. & Marengo, F.D. (2007) . Rapid recovery of releasable vesicles and formation of nonreleasable endosomes follow intense exocytosis in chromaffin cells. American Journal of Physiology - Cell Physiology, 293(5), C1509-C1522.
http://dx.doi.org/10.1152/ajpcell.00632.2006
---------- CHICAGO ----------
Bay, A.E.P., Ibañez, L.I., Marengo, F.D. "Rapid recovery of releasable vesicles and formation of nonreleasable endosomes follow intense exocytosis in chromaffin cells" . American Journal of Physiology - Cell Physiology 293, no. 5 (2007) : C1509-C1522.
http://dx.doi.org/10.1152/ajpcell.00632.2006
---------- MLA ----------
Bay, A.E.P., Ibañez, L.I., Marengo, F.D. "Rapid recovery of releasable vesicles and formation of nonreleasable endosomes follow intense exocytosis in chromaffin cells" . American Journal of Physiology - Cell Physiology, vol. 293, no. 5, 2007, pp. C1509-C1522.
http://dx.doi.org/10.1152/ajpcell.00632.2006
---------- VANCOUVER ----------
Bay, A.E.P., Ibañez, L.I., Marengo, F.D. Rapid recovery of releasable vesicles and formation of nonreleasable endosomes follow intense exocytosis in chromaffin cells. Am. J. Physiol. Cell Physiol. 2007;293(5):C1509-C1522.
http://dx.doi.org/10.1152/ajpcell.00632.2006