Regulation of Endoplasmic Reticulum–Mitochondria Tethering and Ca<sup>2+</sup> Fluxes by TDP-43 via GSK3β

Mitochondria–ER contacts (MERCs), tightly regulated by numerous tethering proteins that act as molecular and functional connections between the two organelles, are essential to maintain a variety of cellular functions. Such contacts are often compromised in the early stages of many neurodegenerative...

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Autores principales:	Caterina Peggion, Maria Lina Massimino, Raphael Severino Bonadio, Federica Lia, Raffaele Lopreiato, Stefano Cagnin, Tito Calì, Alessandro Bertoli
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	amyotrophic lateral sclerosis TDP-43 ER–mitochondria contacts calcium homeostasis SPLICS neurodegenerative disorders Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/f39a3ef2d05c491fa6beb720a4d37a92
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spelling	oai:doaj.org-article:f39a3ef2d05c491fa6beb720a4d37a922021-11-11T17:16:46ZRegulation of Endoplasmic Reticulum–Mitochondria Tethering and Ca<sup>2+</sup> Fluxes by TDP-43 via GSK3β10.3390/ijms2221118531422-00671661-6596https://doaj.org/article/f39a3ef2d05c491fa6beb720a4d37a922021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11853https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Mitochondria–ER contacts (MERCs), tightly regulated by numerous tethering proteins that act as molecular and functional connections between the two organelles, are essential to maintain a variety of cellular functions. Such contacts are often compromised in the early stages of many neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS). TDP-43, a nuclear protein mainly involved in RNA metabolism, has been repeatedly associated with ALS pathogenesis and other neurodegenerative diseases. Although TDP-43 neuropathological mechanisms are still unclear, the accumulation of the protein in cytoplasmic inclusions may underlie a protein loss-of-function effect. Accordingly, we investigated the impact of siRNA-mediated TDP-43 silencing on MERCs and the related cellular parameters in HeLa cells using GFP-based probes for MERCs quantification and aequorin-based probes for local Ca<sup>2+</sup> measurements, combined with targeted protein and mRNA profiling. Our results demonstrated that TDP-43 down-regulation decreases MERCs density, thereby remarkably reducing mitochondria Ca<sup>2+</sup> uptake after ER Ca<sup>2+</sup> release. Thorough mRNA and protein analyses did not highlight altered expression of proteins involved in MERCs assembly or Ca<sup>2+</sup>-mediated ER–mitochondria cross-talk, nor alterations of mitochondrial density and morphology were observed by confocal microscopy. Further mechanistic inspections, however, suggested that the observed cellular alterations are correlated to increased expression/activity of GSK3β, previously associated with MERCs disruption.Caterina PeggionMaria Lina MassiminoRaphael Severino BonadioFederica LiaRaffaele LopreiatoStefano CagninTito CalìAlessandro BertoliMDPI AGarticleamyotrophic lateral sclerosisTDP-43ER–mitochondria contactscalcium homeostasisSPLICSneurodegenerative disordersBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11853, p 11853 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	amyotrophic lateral sclerosis TDP-43 ER–mitochondria contacts calcium homeostasis SPLICS neurodegenerative disorders Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	amyotrophic lateral sclerosis TDP-43 ER–mitochondria contacts calcium homeostasis SPLICS neurodegenerative disorders Biology (General) QH301-705.5 Chemistry QD1-999 Caterina Peggion Maria Lina Massimino Raphael Severino Bonadio Federica Lia Raffaele Lopreiato Stefano Cagnin Tito Calì Alessandro Bertoli Regulation of Endoplasmic Reticulum–Mitochondria Tethering and Ca<sup>2+</sup> Fluxes by TDP-43 via GSK3β
description	Mitochondria–ER contacts (MERCs), tightly regulated by numerous tethering proteins that act as molecular and functional connections between the two organelles, are essential to maintain a variety of cellular functions. Such contacts are often compromised in the early stages of many neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS). TDP-43, a nuclear protein mainly involved in RNA metabolism, has been repeatedly associated with ALS pathogenesis and other neurodegenerative diseases. Although TDP-43 neuropathological mechanisms are still unclear, the accumulation of the protein in cytoplasmic inclusions may underlie a protein loss-of-function effect. Accordingly, we investigated the impact of siRNA-mediated TDP-43 silencing on MERCs and the related cellular parameters in HeLa cells using GFP-based probes for MERCs quantification and aequorin-based probes for local Ca<sup>2+</sup> measurements, combined with targeted protein and mRNA profiling. Our results demonstrated that TDP-43 down-regulation decreases MERCs density, thereby remarkably reducing mitochondria Ca<sup>2+</sup> uptake after ER Ca<sup>2+</sup> release. Thorough mRNA and protein analyses did not highlight altered expression of proteins involved in MERCs assembly or Ca<sup>2+</sup>-mediated ER–mitochondria cross-talk, nor alterations of mitochondrial density and morphology were observed by confocal microscopy. Further mechanistic inspections, however, suggested that the observed cellular alterations are correlated to increased expression/activity of GSK3β, previously associated with MERCs disruption.
format	article
author	Caterina Peggion Maria Lina Massimino Raphael Severino Bonadio Federica Lia Raffaele Lopreiato Stefano Cagnin Tito Calì Alessandro Bertoli
author_facet	Caterina Peggion Maria Lina Massimino Raphael Severino Bonadio Federica Lia Raffaele Lopreiato Stefano Cagnin Tito Calì Alessandro Bertoli
author_sort	Caterina Peggion
title	Regulation of Endoplasmic Reticulum–Mitochondria Tethering and Ca<sup>2+</sup> Fluxes by TDP-43 via GSK3β
title_short	Regulation of Endoplasmic Reticulum–Mitochondria Tethering and Ca<sup>2+</sup> Fluxes by TDP-43 via GSK3β
title_full	Regulation of Endoplasmic Reticulum–Mitochondria Tethering and Ca<sup>2+</sup> Fluxes by TDP-43 via GSK3β
title_fullStr	Regulation of Endoplasmic Reticulum–Mitochondria Tethering and Ca<sup>2+</sup> Fluxes by TDP-43 via GSK3β
title_full_unstemmed	Regulation of Endoplasmic Reticulum–Mitochondria Tethering and Ca<sup>2+</sup> Fluxes by TDP-43 via GSK3β
title_sort	regulation of endoplasmic reticulum–mitochondria tethering and ca<sup>2+</sup> fluxes by tdp-43 via gsk3β
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/f39a3ef2d05c491fa6beb720a4d37a92
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Regulation of Endoplasmic Reticulum–Mitochondria Tethering and Ca<sup>2+</sup> Fluxes by TDP-43 via GSK3β

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