Membrane cycling after the excess retrieval mode of rapid endocytosis in mouse chromaffin cells
Aim: After exocytosis, neuroendocrine cells and neurones keep constant the plasma membrane and the releasable vesicle pools by performing endocytosis and vesicular cycling. Patch-clamp capacitance measurements on chromaffin cells showed that strong Ca +2 entry activates excess retrieval: a rapid end...
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todo:paper_17481708_v204_n3_p403_PerezBay2023-10-03T16:32:16Z Membrane cycling after the excess retrieval mode of rapid endocytosis in mouse chromaffin cells Perez Bay, A.E. Belingheri, A.V. Álvarez, Y.D. Marengo, F.D. Calcium signal Endocytosis Exocytosis FM1-43 Membrane cycling calcium calcium channel L type cholinergic receptor stimulating agent dextran fm 143 potassium protein kinase C unclassified drug animal cell animal experiment article calcium cell level calcium signaling calcium transport cell assay cell membrane cell surface cellular distribution chromaffin cell controlled study cytosol endocytosis endoplasmic reticulum exocytosis fluorescence imaging lysosome mouse nonhuman priority journal recycling Animals Calcium Calcium Channel Blockers Calcium Channels, L-Type Calcium Signaling Cell Membrane Cholinergic Agonists Chromaffin Cells Electric Capacitance Endocytosis Endoplasmic Reticulum Endosomes Exocytosis Fluorescent Dyes Membrane Fusion Membrane Potentials Mice Microscopy, Fluorescence Patch-Clamp Techniques Potassium Protein Kinase C Protein Kinase Inhibitors Pyridinium Compounds Quaternary Ammonium Compounds Time Factors Aim: After exocytosis, neuroendocrine cells and neurones keep constant the plasma membrane and the releasable vesicle pools by performing endocytosis and vesicular cycling. Patch-clamp capacitance measurements on chromaffin cells showed that strong Ca +2 entry activates excess retrieval: a rapid endocytosis process that retrieves more membrane than the one fused by preceding exocytosis. The main purpose of the present experiments was to study the recycling pathway that follows excess retrieval, which is unknown. Methods: Membrane recycling after exocytosis-endocytosis can be studied by fluorescence imaging assays with FM1-43 (Perez Bay et al. Am J Physiol Cell Physiol 2007; 293, C1509). In this work, we used this assay in combination with fluorescent dextrans and specific organelle-targeted antibodies to study the membrane recycling after excess retrieval in mouse chromaffin cells. Results: Excess retrieval was observed after the application of high-K + or cholinergic agonists during 15 or 30s in the presence of FM1-43. We found that the excess retrieval membrane pool (defined as endocytosis-exocytosis) was associated with the generation of a non-releasable fraction of membrane (up to 30% of plasma membrane surface) colocalizing with the lysosomal compartment. The excess retrieval membrane pool followed a saturable cytosolic Ca 2+ dependency, and it was suppressed by inhibitors of L-type Ca 2+ channels, endoplasmic reticulum Ca 2+ release and PKC. Conclusion: Excess retrieval is not associated with the cycling of releasable vesicles, but it is related to the formation of non-releasable endosomes. This process is activated by a concerted contribution of Ca 2+ entry through L-channels and Ca 2+ release from endoplasmic reticulum. © 2011 The Authors. Acta Physiologica © 2011 Scandinavian Physiological Society. Fil:Perez Bay, A.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Belingheri, A.V. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Álvarez, Y.D. 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_17481708_v204_n3_p403_PerezBay |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Calcium signal Endocytosis Exocytosis FM1-43 Membrane cycling calcium calcium channel L type cholinergic receptor stimulating agent dextran fm 143 potassium protein kinase C unclassified drug animal cell animal experiment article calcium cell level calcium signaling calcium transport cell assay cell membrane cell surface cellular distribution chromaffin cell controlled study cytosol endocytosis endoplasmic reticulum exocytosis fluorescence imaging lysosome mouse nonhuman priority journal recycling Animals Calcium Calcium Channel Blockers Calcium Channels, L-Type Calcium Signaling Cell Membrane Cholinergic Agonists Chromaffin Cells Electric Capacitance Endocytosis Endoplasmic Reticulum Endosomes Exocytosis Fluorescent Dyes Membrane Fusion Membrane Potentials Mice Microscopy, Fluorescence Patch-Clamp Techniques Potassium Protein Kinase C Protein Kinase Inhibitors Pyridinium Compounds Quaternary Ammonium Compounds Time Factors |
| spellingShingle |
Calcium signal Endocytosis Exocytosis FM1-43 Membrane cycling calcium calcium channel L type cholinergic receptor stimulating agent dextran fm 143 potassium protein kinase C unclassified drug animal cell animal experiment article calcium cell level calcium signaling calcium transport cell assay cell membrane cell surface cellular distribution chromaffin cell controlled study cytosol endocytosis endoplasmic reticulum exocytosis fluorescence imaging lysosome mouse nonhuman priority journal recycling Animals Calcium Calcium Channel Blockers Calcium Channels, L-Type Calcium Signaling Cell Membrane Cholinergic Agonists Chromaffin Cells Electric Capacitance Endocytosis Endoplasmic Reticulum Endosomes Exocytosis Fluorescent Dyes Membrane Fusion Membrane Potentials Mice Microscopy, Fluorescence Patch-Clamp Techniques Potassium Protein Kinase C Protein Kinase Inhibitors Pyridinium Compounds Quaternary Ammonium Compounds Time Factors Perez Bay, A.E. Belingheri, A.V. Álvarez, Y.D. Marengo, F.D. Membrane cycling after the excess retrieval mode of rapid endocytosis in mouse chromaffin cells |
| topic_facet |
Calcium signal Endocytosis Exocytosis FM1-43 Membrane cycling calcium calcium channel L type cholinergic receptor stimulating agent dextran fm 143 potassium protein kinase C unclassified drug animal cell animal experiment article calcium cell level calcium signaling calcium transport cell assay cell membrane cell surface cellular distribution chromaffin cell controlled study cytosol endocytosis endoplasmic reticulum exocytosis fluorescence imaging lysosome mouse nonhuman priority journal recycling Animals Calcium Calcium Channel Blockers Calcium Channels, L-Type Calcium Signaling Cell Membrane Cholinergic Agonists Chromaffin Cells Electric Capacitance Endocytosis Endoplasmic Reticulum Endosomes Exocytosis Fluorescent Dyes Membrane Fusion Membrane Potentials Mice Microscopy, Fluorescence Patch-Clamp Techniques Potassium Protein Kinase C Protein Kinase Inhibitors Pyridinium Compounds Quaternary Ammonium Compounds Time Factors |
| description |
Aim: After exocytosis, neuroendocrine cells and neurones keep constant the plasma membrane and the releasable vesicle pools by performing endocytosis and vesicular cycling. Patch-clamp capacitance measurements on chromaffin cells showed that strong Ca +2 entry activates excess retrieval: a rapid endocytosis process that retrieves more membrane than the one fused by preceding exocytosis. The main purpose of the present experiments was to study the recycling pathway that follows excess retrieval, which is unknown. Methods: Membrane recycling after exocytosis-endocytosis can be studied by fluorescence imaging assays with FM1-43 (Perez Bay et al. Am J Physiol Cell Physiol 2007; 293, C1509). In this work, we used this assay in combination with fluorescent dextrans and specific organelle-targeted antibodies to study the membrane recycling after excess retrieval in mouse chromaffin cells. Results: Excess retrieval was observed after the application of high-K + or cholinergic agonists during 15 or 30s in the presence of FM1-43. We found that the excess retrieval membrane pool (defined as endocytosis-exocytosis) was associated with the generation of a non-releasable fraction of membrane (up to 30% of plasma membrane surface) colocalizing with the lysosomal compartment. The excess retrieval membrane pool followed a saturable cytosolic Ca 2+ dependency, and it was suppressed by inhibitors of L-type Ca 2+ channels, endoplasmic reticulum Ca 2+ release and PKC. Conclusion: Excess retrieval is not associated with the cycling of releasable vesicles, but it is related to the formation of non-releasable endosomes. This process is activated by a concerted contribution of Ca 2+ entry through L-channels and Ca 2+ release from endoplasmic reticulum. © 2011 The Authors. Acta Physiologica © 2011 Scandinavian Physiological Society. |
| format |
JOUR |
| author |
Perez Bay, A.E. Belingheri, A.V. Álvarez, Y.D. Marengo, F.D. |
| author_facet |
Perez Bay, A.E. Belingheri, A.V. Álvarez, Y.D. Marengo, F.D. |
| author_sort |
Perez Bay, A.E. |
| title |
Membrane cycling after the excess retrieval mode of rapid endocytosis in mouse chromaffin cells |
| title_short |
Membrane cycling after the excess retrieval mode of rapid endocytosis in mouse chromaffin cells |
| title_full |
Membrane cycling after the excess retrieval mode of rapid endocytosis in mouse chromaffin cells |
| title_fullStr |
Membrane cycling after the excess retrieval mode of rapid endocytosis in mouse chromaffin cells |
| title_full_unstemmed |
Membrane cycling after the excess retrieval mode of rapid endocytosis in mouse chromaffin cells |
| title_sort |
membrane cycling after the excess retrieval mode of rapid endocytosis in mouse chromaffin cells |
| url |
http://hdl.handle.net/20.500.12110/paper_17481708_v204_n3_p403_PerezBay |
| work_keys_str_mv |
AT perezbayae membranecyclingaftertheexcessretrievalmodeofrapidendocytosisinmousechromaffincells AT belingheriav membranecyclingaftertheexcessretrievalmodeofrapidendocytosisinmousechromaffincells AT alvarezyd membranecyclingaftertheexcessretrievalmodeofrapidendocytosisinmousechromaffincells AT marengofd membranecyclingaftertheexcessretrievalmodeofrapidendocytosisinmousechromaffincells |
| _version_ |
1782029034706698240 |