2026-04-27T01:38:30Zhttps://uvadoc.uva.es/oai/request

oai:uvadoc.uva.es:10324/59742024-12-16T12:04:35Zcom_10324_1133com_10324_931com_10324_894col_10324_1209

Moreno Díaz-Calderón, Alfredo Santo Domingo Mayoral, Jaime Fonteriz García, Rosalba Inés Domínguez Lobatón, María Carmen Montero Zoccola, María Teresa Álvarez Martín, Javier 2014-09-16T08:55:31Z 2014-09-16T08:55:31Z 2010 Journal of Structural Biology, 2010, vol. 172, p. 261-269 1047-8477 http://uvadoc.uva.es/handle/10324/5974 10.1016/j.jsb.2010.06.015 261 269 Journal of Structural Biology 172 Producción Científica Secretory vesicles have low pH and have been classically identified as those labelled by a series of acidic fluorescent dyes such as acridine orange or neutral red, which accumulate into the vesicles according to the pH gradient. More recently, several fusion proteins containing enhanced green fluorescent protein (EGFP) and targeted to the secretory vesicles have been engineered. Both targeted fluorescent proteins and acidic dyes have been used, separately or combined, to monitor the dynamics of secretory vesicle movements and their fusion with the plasma membrane. We have now investigated in detail the degree of colocalization of both types of probes using several fusion proteins targeted to the vesicles (synaptobrevin2- EGFP, Cromogranin A-EGFP and neuropeptide Y-EGFP) and several acidic dyes (acridine orange, neutral red and lysotracker red) in chromaffin cells, PC12 cells and GH3 cells. We find that all the acidic dyes labelled the same population of vesicles. However, that population was largely different from the one labelled by the targeted proteins, with very little colocalization among them, in all the cell types studied. Our data show that the vesicles containing the proteins more characteristic of the secretory vesicles are not labelled by the acidic dyes, and vice versa. Peptide glycyl-L-phenylalanine 2-naphthylamide (GPN) produced a rapid and selective disruption of the vesicles labelled by acidic dyes, suggesting that they could be mainly lysosomes. Therefore, these labelling techniques distinguish two clearly different sets of acidic vesicles in neuroendocrine cells. This finding should be taken into account whenever vesicle dynamics is studied using these techniques. application/pdf eng Elsevier Inc. info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-nd/4.0/ Attribution-NonCommercial-NoDerivatives 4.0 International Calcio en el organismo A confocal study on the visualization of chromaffin cell secretory vesicles with fluorescent targeted probes and acidic dyes info:eu-repo/semantics/article SI