Characterization of Stress Granule Protein Turnover in Neuronal Progenitor Cells Using Correlative STED and NanoSIMS Imaging
Stress granules (SGs) are stress-induced biomolecular condensates which originate primarily from inactivated RNA translation machinery and translation initiation factors. SG formation is an important defensive mechanism for cell survival, while its dysfunction has been linked to neurodegenerative di...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/3/2546 |
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| author | Stefania Rabasco Alicia A. Lork Emmanuel Berlin Tho D. K. Nguyen Carl Ernst Nicolas Locker Andrew G. Ewing Nhu T. N. Phan |
| author_facet | Stefania Rabasco Alicia A. Lork Emmanuel Berlin Tho D. K. Nguyen Carl Ernst Nicolas Locker Andrew G. Ewing Nhu T. N. Phan |
| author_sort | Stefania Rabasco |
| collection | DOAJ |
| description | Stress granules (SGs) are stress-induced biomolecular condensates which originate primarily from inactivated RNA translation machinery and translation initiation factors. SG formation is an important defensive mechanism for cell survival, while its dysfunction has been linked to neurodegenerative diseases. However, the molecular mechanisms of SG assembly and disassembly, as well as their impacts on cellular recovery, are not fully understood. More thorough investigations into the molecular dynamics of SG pathways are required to understand the pathophysiological roles of SGs in cellular systems. Here, we characterize the SG and cytoplasmic protein turnover in neuronal progenitor cells (NPCs) under stressed and non-stressed conditions using correlative STED and NanoSIMS imaging. We incubate NPCs with isotopically labelled (<sup>15</sup>N) leucine and stress them with the ER stressor thapsigargin (TG). A correlation of STED and NanoSIMS allows the localization of individual SGs (using STED), and their protein turnover can then be extracted based on the <sup>15</sup>N/<sup>14</sup>N ratio (using NanoSIMS). We found that TG-induced SGs, which are highly dynamic domains, recruit their constituents predominantly from the cytoplasm. Moreover, ER stress impairs the total cellular protein turnover regimen, and this impairment is not restored after the commonly proceeded stress recovery period. |
| first_indexed | 2024-03-11T09:40:31Z |
| format | Article |
| id | doaj.art-0f2d7a356695453baa9bcc12b498b80a |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T09:40:31Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-0f2d7a356695453baa9bcc12b498b80a2023-11-16T16:58:50ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-01-01243254610.3390/ijms24032546Characterization of Stress Granule Protein Turnover in Neuronal Progenitor Cells Using Correlative STED and NanoSIMS ImagingStefania Rabasco0Alicia A. Lork1Emmanuel Berlin2Tho D. K. Nguyen3Carl Ernst4Nicolas Locker5Andrew G. Ewing6Nhu T. N. Phan7Department of Chemistry and Molecular Biology, University of Gothenburg, SE-412 96 Gothenburg, SwedenDepartment of Chemistry and Molecular Biology, University of Gothenburg, SE-412 96 Gothenburg, SwedenDepartment of Chemistry and Molecular Biology, University of Gothenburg, SE-412 96 Gothenburg, SwedenDepartment of Chemistry and Molecular Biology, University of Gothenburg, SE-412 96 Gothenburg, SwedenHuman Genetics, McGill University, Montreal, QC H4H1R3, CanadaFaculty of Health and Medical Sciences, School of Biosciences and Medicine, University of Surrey, Guildford GU2 7XH, UKDepartment of Chemistry and Molecular Biology, University of Gothenburg, SE-412 96 Gothenburg, SwedenDepartment of Chemistry and Molecular Biology, University of Gothenburg, SE-412 96 Gothenburg, SwedenStress granules (SGs) are stress-induced biomolecular condensates which originate primarily from inactivated RNA translation machinery and translation initiation factors. SG formation is an important defensive mechanism for cell survival, while its dysfunction has been linked to neurodegenerative diseases. However, the molecular mechanisms of SG assembly and disassembly, as well as their impacts on cellular recovery, are not fully understood. More thorough investigations into the molecular dynamics of SG pathways are required to understand the pathophysiological roles of SGs in cellular systems. Here, we characterize the SG and cytoplasmic protein turnover in neuronal progenitor cells (NPCs) under stressed and non-stressed conditions using correlative STED and NanoSIMS imaging. We incubate NPCs with isotopically labelled (<sup>15</sup>N) leucine and stress them with the ER stressor thapsigargin (TG). A correlation of STED and NanoSIMS allows the localization of individual SGs (using STED), and their protein turnover can then be extracted based on the <sup>15</sup>N/<sup>14</sup>N ratio (using NanoSIMS). We found that TG-induced SGs, which are highly dynamic domains, recruit their constituents predominantly from the cytoplasm. Moreover, ER stress impairs the total cellular protein turnover regimen, and this impairment is not restored after the commonly proceeded stress recovery period.https://www.mdpi.com/1422-0067/24/3/2546stress granulesNanoSIMSprotein turnoverneuronal progenitor cellsmass spectrometry imaging |
| spellingShingle | Stefania Rabasco Alicia A. Lork Emmanuel Berlin Tho D. K. Nguyen Carl Ernst Nicolas Locker Andrew G. Ewing Nhu T. N. Phan Characterization of Stress Granule Protein Turnover in Neuronal Progenitor Cells Using Correlative STED and NanoSIMS Imaging International Journal of Molecular Sciences stress granules NanoSIMS protein turnover neuronal progenitor cells mass spectrometry imaging |
| title | Characterization of Stress Granule Protein Turnover in Neuronal Progenitor Cells Using Correlative STED and NanoSIMS Imaging |
| title_full | Characterization of Stress Granule Protein Turnover in Neuronal Progenitor Cells Using Correlative STED and NanoSIMS Imaging |
| title_fullStr | Characterization of Stress Granule Protein Turnover in Neuronal Progenitor Cells Using Correlative STED and NanoSIMS Imaging |
| title_full_unstemmed | Characterization of Stress Granule Protein Turnover in Neuronal Progenitor Cells Using Correlative STED and NanoSIMS Imaging |
| title_short | Characterization of Stress Granule Protein Turnover in Neuronal Progenitor Cells Using Correlative STED and NanoSIMS Imaging |
| title_sort | characterization of stress granule protein turnover in neuronal progenitor cells using correlative sted and nanosims imaging |
| topic | stress granules NanoSIMS protein turnover neuronal progenitor cells mass spectrometry imaging |
| url | https://www.mdpi.com/1422-0067/24/3/2546 |
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