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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2022
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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. |
format | Online Article Text |
id | pubmed-8945366 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
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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