Mitochondria Endoplasmic Reticulum Contact Sites (MERCs): Proximity Ligation Assay as a Tool to Study Organelle Interaction

Organelles cooperate with each other to regulate vital cellular homoeostatic functions. This occurs through the formation of close connections through membrane contact sites. Mitochondria-Endoplasmic-Reticulum (ER) contact sites (MERCS) are one of such contact sites that regulate numerous biological...

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Autores principales:	Sara Benhammouda, Anjali Vishwakarma, Priya Gatti, Marc Germain
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	mitochondia endoplasmic reticulum organelle contact sites organelle contact sites methodologies Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/0df2e776b138444e81c10ba5bbc18d53
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spelling	oai:doaj.org-article:0df2e776b138444e81c10ba5bbc18d532021-12-03T06:48:58ZMitochondria Endoplasmic Reticulum Contact Sites (MERCs): Proximity Ligation Assay as a Tool to Study Organelle Interaction2296-634X10.3389/fcell.2021.789959https://doaj.org/article/0df2e776b138444e81c10ba5bbc18d532021-12-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fcell.2021.789959/fullhttps://doaj.org/toc/2296-634XOrganelles cooperate with each other to regulate vital cellular homoeostatic functions. This occurs through the formation of close connections through membrane contact sites. Mitochondria-Endoplasmic-Reticulum (ER) contact sites (MERCS) are one of such contact sites that regulate numerous biological processes by controlling calcium and metabolic homeostasis. However, the extent to which contact sites shape cellular biology and the underlying mechanisms remain to be fully elucidated. A number of biochemical and imaging approaches have been established to address these questions, resulting in the identification of a number of molecular tethers between mitochondria and the ER. Among these techniques, fluorescence-based imaging is widely used, including analysing signal overlap between two organelles and more selective techniques such as in-situ proximity ligation assay (PLA). While these two techniques allow the detection of endogenous proteins, preventing some problems associated with techniques relying on overexpression (FRET, split fluorescence probes), they come with their own issues. In addition, proper image analysis is required to minimise potential artefacts associated with these methods. In this review, we discuss the protocols and outline the limitations of fluorescence-based approaches used to assess MERCs using endogenous proteins.Sara BenhammoudaSara BenhammoudaAnjali VishwakarmaAnjali VishwakarmaPriya GattiPriya GattiMarc GermainMarc GermainFrontiers Media S.A.articlemitochondiaendoplasmic reticulumorganelle contact sitesorganellecontact sites methodologiesBiology (General)QH301-705.5ENFrontiers in Cell and Developmental Biology, Vol 9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	mitochondia endoplasmic reticulum organelle contact sites organelle contact sites methodologies Biology (General) QH301-705.5
spellingShingle	mitochondia endoplasmic reticulum organelle contact sites organelle contact sites methodologies Biology (General) QH301-705.5 Sara Benhammouda Sara Benhammouda Anjali Vishwakarma Anjali Vishwakarma Priya Gatti Priya Gatti Marc Germain Marc Germain Mitochondria Endoplasmic Reticulum Contact Sites (MERCs): Proximity Ligation Assay as a Tool to Study Organelle Interaction
description	Organelles cooperate with each other to regulate vital cellular homoeostatic functions. This occurs through the formation of close connections through membrane contact sites. Mitochondria-Endoplasmic-Reticulum (ER) contact sites (MERCS) are one of such contact sites that regulate numerous biological processes by controlling calcium and metabolic homeostasis. However, the extent to which contact sites shape cellular biology and the underlying mechanisms remain to be fully elucidated. A number of biochemical and imaging approaches have been established to address these questions, resulting in the identification of a number of molecular tethers between mitochondria and the ER. Among these techniques, fluorescence-based imaging is widely used, including analysing signal overlap between two organelles and more selective techniques such as in-situ proximity ligation assay (PLA). While these two techniques allow the detection of endogenous proteins, preventing some problems associated with techniques relying on overexpression (FRET, split fluorescence probes), they come with their own issues. In addition, proper image analysis is required to minimise potential artefacts associated with these methods. In this review, we discuss the protocols and outline the limitations of fluorescence-based approaches used to assess MERCs using endogenous proteins.
format	article
author	Sara Benhammouda Sara Benhammouda Anjali Vishwakarma Anjali Vishwakarma Priya Gatti Priya Gatti Marc Germain Marc Germain
author_facet	Sara Benhammouda Sara Benhammouda Anjali Vishwakarma Anjali Vishwakarma Priya Gatti Priya Gatti Marc Germain Marc Germain
author_sort	Sara Benhammouda
title	Mitochondria Endoplasmic Reticulum Contact Sites (MERCs): Proximity Ligation Assay as a Tool to Study Organelle Interaction
title_short	Mitochondria Endoplasmic Reticulum Contact Sites (MERCs): Proximity Ligation Assay as a Tool to Study Organelle Interaction
title_full	Mitochondria Endoplasmic Reticulum Contact Sites (MERCs): Proximity Ligation Assay as a Tool to Study Organelle Interaction
title_fullStr	Mitochondria Endoplasmic Reticulum Contact Sites (MERCs): Proximity Ligation Assay as a Tool to Study Organelle Interaction
title_full_unstemmed	Mitochondria Endoplasmic Reticulum Contact Sites (MERCs): Proximity Ligation Assay as a Tool to Study Organelle Interaction
title_sort	mitochondria endoplasmic reticulum contact sites (mercs): proximity ligation assay as a tool to study organelle interaction
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/0df2e776b138444e81c10ba5bbc18d53
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Mitochondria Endoplasmic Reticulum Contact Sites (MERCs): Proximity Ligation Assay as a Tool to Study Organelle Interaction

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