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Macropinocytic entry of isolated mitochondria in epidermal growth factor-activated human osteosarcoma cells
Mammalian mitochondria can be transferred between cells both in culture and in vivo. There is evidence that isolated mitochondria enter cells by endocytosis, but the mechanism has not been fully characterised. We investigated the entry mechanism of isolated mitochondria into human osteosarcoma (HOS)...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5634993/ https://www.ncbi.nlm.nih.gov/pubmed/29018288 http://dx.doi.org/10.1038/s41598-017-13227-0 |
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author | Patel, Dipali Rorbach, Joanna Downes, Kate Szukszto, Maciej J. Pekalski, Marcin L. Minczuk, Michal |
author_facet | Patel, Dipali Rorbach, Joanna Downes, Kate Szukszto, Maciej J. Pekalski, Marcin L. Minczuk, Michal |
author_sort | Patel, Dipali |
collection | PubMed |
description | Mammalian mitochondria can be transferred between cells both in culture and in vivo. There is evidence that isolated mitochondria enter cells by endocytosis, but the mechanism has not been fully characterised. We investigated the entry mechanism of isolated mitochondria into human osteosarcoma (HOS) cells. Initially we confirmed that respiratory-competent cells can be produced following incubation of HOS cells lacking mitochondrial DNA (mtDNA) with functional exogenous mitochondria and selection in a restrictive medium. Treatment of HOS cells with inhibitors of different endocytic pathways suggest that uptake of EGFP-labelled mitochondria occurs via an actin-dependent endocytic pathway which is consistent with macropinocytosis. We later utilised time-lapse microscopy to show that internalised mitochondria were found in large, motile cellular vesicles. Finally, we used confocal imaging to show that EGFP-labelled mitochondria colocalise with a macropinocytic cargo molecule during internalisation, HOS cells produce membrane ruffles interacting with external mitochondria during uptake and EGFP-labelled mitochondria are found within early macropinosomes inside cells. In conclusion our results are consistent with isolated mitochondria being internalised by macropinocytosis in HOS cells. |
format | Online Article Text |
id | pubmed-5634993 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-56349932017-10-18 Macropinocytic entry of isolated mitochondria in epidermal growth factor-activated human osteosarcoma cells Patel, Dipali Rorbach, Joanna Downes, Kate Szukszto, Maciej J. Pekalski, Marcin L. Minczuk, Michal Sci Rep Article Mammalian mitochondria can be transferred between cells both in culture and in vivo. There is evidence that isolated mitochondria enter cells by endocytosis, but the mechanism has not been fully characterised. We investigated the entry mechanism of isolated mitochondria into human osteosarcoma (HOS) cells. Initially we confirmed that respiratory-competent cells can be produced following incubation of HOS cells lacking mitochondrial DNA (mtDNA) with functional exogenous mitochondria and selection in a restrictive medium. Treatment of HOS cells with inhibitors of different endocytic pathways suggest that uptake of EGFP-labelled mitochondria occurs via an actin-dependent endocytic pathway which is consistent with macropinocytosis. We later utilised time-lapse microscopy to show that internalised mitochondria were found in large, motile cellular vesicles. Finally, we used confocal imaging to show that EGFP-labelled mitochondria colocalise with a macropinocytic cargo molecule during internalisation, HOS cells produce membrane ruffles interacting with external mitochondria during uptake and EGFP-labelled mitochondria are found within early macropinosomes inside cells. In conclusion our results are consistent with isolated mitochondria being internalised by macropinocytosis in HOS cells. Nature Publishing Group UK 2017-10-10 /pmc/articles/PMC5634993/ /pubmed/29018288 http://dx.doi.org/10.1038/s41598-017-13227-0 Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Patel, Dipali Rorbach, Joanna Downes, Kate Szukszto, Maciej J. Pekalski, Marcin L. Minczuk, Michal Macropinocytic entry of isolated mitochondria in epidermal growth factor-activated human osteosarcoma cells |
title | Macropinocytic entry of isolated mitochondria in epidermal growth factor-activated human osteosarcoma cells |
title_full | Macropinocytic entry of isolated mitochondria in epidermal growth factor-activated human osteosarcoma cells |
title_fullStr | Macropinocytic entry of isolated mitochondria in epidermal growth factor-activated human osteosarcoma cells |
title_full_unstemmed | Macropinocytic entry of isolated mitochondria in epidermal growth factor-activated human osteosarcoma cells |
title_short | Macropinocytic entry of isolated mitochondria in epidermal growth factor-activated human osteosarcoma cells |
title_sort | macropinocytic entry of isolated mitochondria in epidermal growth factor-activated human osteosarcoma cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5634993/ https://www.ncbi.nlm.nih.gov/pubmed/29018288 http://dx.doi.org/10.1038/s41598-017-13227-0 |
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