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Abstract:

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.

Registro:

Documento: Artículo
Título:Membrane cycling after the excess retrieval mode of rapid endocytosis in mouse chromaffin cells
Autor:Perez Bay, A.E.; Belingheri, A.V.; Álvarez, Y.D.; Marengo, F.D.
Filiación:Laboratorio de Fisiología y Biología Molecular, Universidad de Buenos Aires, Departamento de Fisiología y Biología Molecular y Celular, Inst. de Fisiologia Biologia Molecular y Neurociencias CONICET, Fac. de Ciencias Exactas y Naturales, Buenos Aires, Argentina
Department of Ophthalmology, Dyson Vision Research Institute, Weill Medical College, Cornell University, New York, NY, United States
Palabras clave: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
Año:2012
Volumen:204
Número:3
Página de inicio:403
Página de fin:418
DOI: http://dx.doi.org/10.1111/j.1748-1716.2011.02340.x
Título revista:Acta Physiologica
Título revista abreviado:Acta Physiol.
ISSN:17481708
CAS:calcium, 14092-94-5, 7440-70-2; dextran, 87915-38-6, 9014-78-2; potassium, 7440-09-7; protein kinase C, 141436-78-4; Calcium, 7440-70-2; Calcium Channel Blockers; Calcium Channels, L-Type; Cholinergic Agonists; FM1 43; Fluorescent Dyes; Potassium, 7440-09-7; Protein Kinase C, 2.7.11.13; Protein Kinase Inhibitors; Pyridinium Compounds; Quaternary Ammonium Compounds
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_17481708_v204_n3_p403_PerezBay

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Citas:

---------- APA ----------
Perez Bay, A.E., Belingheri, A.V., Álvarez, Y.D. & Marengo, F.D. (2012) . Membrane cycling after the excess retrieval mode of rapid endocytosis in mouse chromaffin cells. Acta Physiologica, 204(3), 403-418.
http://dx.doi.org/10.1111/j.1748-1716.2011.02340.x
---------- CHICAGO ----------
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" . Acta Physiologica 204, no. 3 (2012) : 403-418.
http://dx.doi.org/10.1111/j.1748-1716.2011.02340.x
---------- MLA ----------
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" . Acta Physiologica, vol. 204, no. 3, 2012, pp. 403-418.
http://dx.doi.org/10.1111/j.1748-1716.2011.02340.x
---------- VANCOUVER ----------
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. Acta Physiol. 2012;204(3):403-418.
http://dx.doi.org/10.1111/j.1748-1716.2011.02340.x