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Intracellular dynamics of the Sigma-1 receptor observed with super-resolution imaging microscopy

Sigma-1 receptor (Sig1R) is an endoplasmic reticulum (ER)-related membrane protein, that forms heteromers with other cellular proteins. As the mechanism of action of this chaperone protein remains unclear, the aim of the present study was to detect and analyze the intracellular dynamics of Sig1R in...

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Autores principales: Kopanchuk, Sergei, Vavers, Edijs, Veiksina, Santa, Ligi, Kadri, Zvejniece, Liga, Dambrova, Maija, Rinken, Ago
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9116656/
https://www.ncbi.nlm.nih.gov/pubmed/35584184
http://dx.doi.org/10.1371/journal.pone.0268563
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author Kopanchuk, Sergei
Vavers, Edijs
Veiksina, Santa
Ligi, Kadri
Zvejniece, Liga
Dambrova, Maija
Rinken, Ago
author_facet Kopanchuk, Sergei
Vavers, Edijs
Veiksina, Santa
Ligi, Kadri
Zvejniece, Liga
Dambrova, Maija
Rinken, Ago
author_sort Kopanchuk, Sergei
collection PubMed
description Sigma-1 receptor (Sig1R) is an endoplasmic reticulum (ER)-related membrane protein, that forms heteromers with other cellular proteins. As the mechanism of action of this chaperone protein remains unclear, the aim of the present study was to detect and analyze the intracellular dynamics of Sig1R in live cells using super-resolution imaging microscopy. For that, the Sig1R-yellow fluorescent protein conjugate (Sig1R-YFP) together with fluorescent markers of cell organelles were transfected into human ovarian adenocarcinoma (SK-OV-3) cells with BacMam technology. Sig1R-YFP was found to be located mainly in the nuclear envelope and in both tubular and vesicular structures of the ER but was not detected in the plasma membrane, even after activation of Sig1R with agonists. The super-resolution radial fluctuations approach (SRRF) performed with a highly inclined and laminated optical sheet (HILO) fluorescence microscope indicated substantial overlap of Sig1R-YFP spots with KDEL-mRFP, slight overlap with pmKate2-mito and no overlap with the markers of endosomes, peroxisomes, lysosomes, or caveolae. Activation of Sig1R with (+)-pentazocine caused a time-dependent decrease in the overlap between Sig1R-YFP and KDEL-mRFP, indicating that the activation of Sig1R decreases its colocalization with the marker of vesicular ER and does not cause comprehensive translocations of Sig1R in cells.
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spelling pubmed-91166562022-05-19 Intracellular dynamics of the Sigma-1 receptor observed with super-resolution imaging microscopy Kopanchuk, Sergei Vavers, Edijs Veiksina, Santa Ligi, Kadri Zvejniece, Liga Dambrova, Maija Rinken, Ago PLoS One Research Article Sigma-1 receptor (Sig1R) is an endoplasmic reticulum (ER)-related membrane protein, that forms heteromers with other cellular proteins. As the mechanism of action of this chaperone protein remains unclear, the aim of the present study was to detect and analyze the intracellular dynamics of Sig1R in live cells using super-resolution imaging microscopy. For that, the Sig1R-yellow fluorescent protein conjugate (Sig1R-YFP) together with fluorescent markers of cell organelles were transfected into human ovarian adenocarcinoma (SK-OV-3) cells with BacMam technology. Sig1R-YFP was found to be located mainly in the nuclear envelope and in both tubular and vesicular structures of the ER but was not detected in the plasma membrane, even after activation of Sig1R with agonists. The super-resolution radial fluctuations approach (SRRF) performed with a highly inclined and laminated optical sheet (HILO) fluorescence microscope indicated substantial overlap of Sig1R-YFP spots with KDEL-mRFP, slight overlap with pmKate2-mito and no overlap with the markers of endosomes, peroxisomes, lysosomes, or caveolae. Activation of Sig1R with (+)-pentazocine caused a time-dependent decrease in the overlap between Sig1R-YFP and KDEL-mRFP, indicating that the activation of Sig1R decreases its colocalization with the marker of vesicular ER and does not cause comprehensive translocations of Sig1R in cells. Public Library of Science 2022-05-18 /pmc/articles/PMC9116656/ /pubmed/35584184 http://dx.doi.org/10.1371/journal.pone.0268563 Text en © 2022 Kopanchuk et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Kopanchuk, Sergei
Vavers, Edijs
Veiksina, Santa
Ligi, Kadri
Zvejniece, Liga
Dambrova, Maija
Rinken, Ago
Intracellular dynamics of the Sigma-1 receptor observed with super-resolution imaging microscopy
title Intracellular dynamics of the Sigma-1 receptor observed with super-resolution imaging microscopy
title_full Intracellular dynamics of the Sigma-1 receptor observed with super-resolution imaging microscopy
title_fullStr Intracellular dynamics of the Sigma-1 receptor observed with super-resolution imaging microscopy
title_full_unstemmed Intracellular dynamics of the Sigma-1 receptor observed with super-resolution imaging microscopy
title_short Intracellular dynamics of the Sigma-1 receptor observed with super-resolution imaging microscopy
title_sort intracellular dynamics of the sigma-1 receptor observed with super-resolution imaging microscopy
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9116656/
https://www.ncbi.nlm.nih.gov/pubmed/35584184
http://dx.doi.org/10.1371/journal.pone.0268563
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