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Visualization of Partial Exocytotic Content Release and Chemical Transport into Nanovesicles in Cells

[Image: see text] For decades, “all-or-none” and “kiss-and-run” were thought to be the only major exocytotic release modes in cell-to-cell communication, while the significance of partial release has not yet been widely recognized and accepted owing to the lack of direct evidence for exocytotic part...

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Autores principales: Nguyen, Tho Duc Khanh, Mellander, Lisa, Lork, Alicia, Thomen, Aurélien, Philipsen, Mai, Kurczy, Michael E., Phan, Nhu T.N., Ewing, Andrew G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2022
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8945366/
https://www.ncbi.nlm.nih.gov/pubmed/35189057
http://dx.doi.org/10.1021/acsnano.2c00344
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author Nguyen, Tho Duc Khanh
Mellander, Lisa
Lork, Alicia
Thomen, Aurélien
Philipsen, Mai
Kurczy, Michael E.
Phan, Nhu T.N.
Ewing, Andrew G.
author_facet Nguyen, Tho Duc Khanh
Mellander, Lisa
Lork, Alicia
Thomen, Aurélien
Philipsen, Mai
Kurczy, Michael E.
Phan, Nhu T.N.
Ewing, Andrew G.
author_sort Nguyen, Tho Duc Khanh
collection PubMed
description [Image: see text] For decades, “all-or-none” and “kiss-and-run” were thought to be the only major exocytotic release modes in cell-to-cell communication, while the significance of partial release has not yet been widely recognized and accepted owing to the lack of direct evidence for exocytotic partial release. Correlative imaging with transmission electron microscopy and NanoSIMS imaging and a dual stable isotope labeling approach was used to study the cargo status of vesicles before and after exocytosis; demonstrating a measurable loss of transmitter in individual vesicles following stimulation due to partial release. Model secretory cells were incubated with (13)C-labeled l-3,4-dihydroxyphenylalanine, resulting in the loading of (13)C-labeled dopamine into their vesicles. A second label, di-N-desethylamiodarone, having the stable isotope (127)I, was introduced during stimulation. A significant drop in the level of (13)C-labeled dopamine and a reduction in vesicle size, with an increasing level of (127)I(–), was observed in vesicles of stimulated cells. Colocalization of (13)C and (127)I(–) in several vesicles was observed after stimulation. Thus, chemical visualization shows transient opening of vesicles to the exterior of the cell without full release the dopamine cargo. We present a direct calculation for the fraction of neurotransmitter release from combined imaging data. The average vesicular release is 60% of the total catecholamine. An important observation is that extracellular molecules can be introduced to cells during the partial exocytotic release process. This nonendocytic transport process appears to be a general route of entry that might be exploited pharmacologically.
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spelling pubmed-89453662022-03-28 Visualization of Partial Exocytotic Content Release and Chemical Transport into Nanovesicles in Cells Nguyen, Tho Duc Khanh Mellander, Lisa Lork, Alicia Thomen, Aurélien Philipsen, Mai Kurczy, Michael E. Phan, Nhu T.N. Ewing, Andrew G. ACS Nano [Image: see text] For decades, “all-or-none” and “kiss-and-run” were thought to be the only major exocytotic release modes in cell-to-cell communication, while the significance of partial release has not yet been widely recognized and accepted owing to the lack of direct evidence for exocytotic partial release. Correlative imaging with transmission electron microscopy and NanoSIMS imaging and a dual stable isotope labeling approach was used to study the cargo status of vesicles before and after exocytosis; demonstrating a measurable loss of transmitter in individual vesicles following stimulation due to partial release. Model secretory cells were incubated with (13)C-labeled l-3,4-dihydroxyphenylalanine, resulting in the loading of (13)C-labeled dopamine into their vesicles. A second label, di-N-desethylamiodarone, having the stable isotope (127)I, was introduced during stimulation. A significant drop in the level of (13)C-labeled dopamine and a reduction in vesicle size, with an increasing level of (127)I(–), was observed in vesicles of stimulated cells. Colocalization of (13)C and (127)I(–) in several vesicles was observed after stimulation. Thus, chemical visualization shows transient opening of vesicles to the exterior of the cell without full release the dopamine cargo. We present a direct calculation for the fraction of neurotransmitter release from combined imaging data. The average vesicular release is 60% of the total catecholamine. An important observation is that extracellular molecules can be introduced to cells during the partial exocytotic release process. This nonendocytic transport process appears to be a general route of entry that might be exploited pharmacologically. American Chemical Society 2022-02-21 2022-03-22 /pmc/articles/PMC8945366/ /pubmed/35189057 http://dx.doi.org/10.1021/acsnano.2c00344 Text en © 2022 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Nguyen, Tho Duc Khanh
Mellander, Lisa
Lork, Alicia
Thomen, Aurélien
Philipsen, Mai
Kurczy, Michael E.
Phan, Nhu T.N.
Ewing, Andrew G.
Visualization of Partial Exocytotic Content Release and Chemical Transport into Nanovesicles in Cells
title Visualization of Partial Exocytotic Content Release and Chemical Transport into Nanovesicles in Cells
title_full Visualization of Partial Exocytotic Content Release and Chemical Transport into Nanovesicles in Cells
title_fullStr Visualization of Partial Exocytotic Content Release and Chemical Transport into Nanovesicles in Cells
title_full_unstemmed Visualization of Partial Exocytotic Content Release and Chemical Transport into Nanovesicles in Cells
title_short Visualization of Partial Exocytotic Content Release and Chemical Transport into Nanovesicles in Cells
title_sort visualization of partial exocytotic content release and chemical transport into nanovesicles in cells
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8945366/
https://www.ncbi.nlm.nih.gov/pubmed/35189057
http://dx.doi.org/10.1021/acsnano.2c00344
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