Rapid recovery of releasable vesicles and formation of nonreleasable endosomes follow intense exocytosis in chromaffin cells
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 ev...
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todo:paper_03636143_v293_n5_pC1509_Bay2023-10-03T15:27:32Z Rapid recovery of releasable vesicles and formation of nonreleasable endosomes follow intense exocytosis in chromaffin cells Bay, A.E.P. Ibañez, L.I. Marengo, F.D. 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 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. Fil:Bay, A.E.P. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Ibañez, L.I. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Marengo, F.D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_03636143_v293_n5_pC1509_Bay |
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Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
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Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
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 |
| spellingShingle |
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 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 |
| topic_facet |
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 |
| description |
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. |
| format |
JOUR |
| author |
Bay, A.E.P. Ibañez, L.I. Marengo, F.D. |
| author_facet |
Bay, A.E.P. Ibañez, L.I. Marengo, F.D. |
| author_sort |
Bay, A.E.P. |
| title |
Rapid recovery of releasable vesicles and formation of nonreleasable endosomes follow intense exocytosis in chromaffin cells |
| title_short |
Rapid recovery of releasable vesicles and formation of nonreleasable endosomes follow intense exocytosis in chromaffin cells |
| title_full |
Rapid recovery of releasable vesicles and formation of nonreleasable endosomes follow intense exocytosis in chromaffin cells |
| title_fullStr |
Rapid recovery of releasable vesicles and formation of nonreleasable endosomes follow intense exocytosis in chromaffin cells |
| title_full_unstemmed |
Rapid recovery of releasable vesicles and formation of nonreleasable endosomes follow intense exocytosis in chromaffin cells |
| title_sort |
rapid recovery of releasable vesicles and formation of nonreleasable endosomes follow intense exocytosis in chromaffin cells |
| url |
http://hdl.handle.net/20.500.12110/paper_03636143_v293_n5_pC1509_Bay |
| work_keys_str_mv |
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