ArticleScientific reports2023
Increased intracellular crowding during hyperosmotic stress.
Article in Scientific reports, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 29 citations in OpenAlex.
- Mapping Protein Diffusion from the Plasma Membrane to the Nucleus: Insights from Fluorescence Correlation Spectroscopy.Biomolecules · 2026Review
- Understanding Osmolality as a Critical Process Parameter in Mammalian Muscle Cell Culture and Biomanufacturing.ACS omega · 2026Review
- Anisosmotic Modulation of Mutant Huntingtin Aggregation vis-a-vis HSP70 Induction─Implications for Aging, Hypo-Hydration, and Neurodegeneration.ACS chemical neuroscience · 2026Article
- Cytoplasmic crowding acts as a porous medium reducing macromolecule diffusion.Proceedings of the National Academy of Sciences of the United States of America · 2026Article
- Rapid activation of p62 body-mediated autophagy in human cells under hyperosmotic stress.Communications biology · 2026Article
- Tissue regenerative medicine: Clinical advances, challenges, and opportunities.APL bioengineering · 2025Article
- High Molecular Crowding in Repair Foci Surrounding DNA Breaks, Measured by Fluorescence Lifetime Imaging Microscopy.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2025Article
- Liquid-Liquid Phase Separation from the Viewpoint of Molecular Crowding Environment: A Raman Imaging Study.The journal of physical chemistry. B · 2025Review
- Microscopy-based techniques for studying the material properties of biomolecular condensates in the cellular environment.Biophysical reviews · 2025Review
- Celebrating 50 years of fluorescence correlation spectroscopy (FCS): Advancing live-cell massively parallel FCS studies with photostable GFPs, mStayGold and StayGold/E138D.Biochimica et biophysica acta. General subjects · 2025Article
- Trans-scale live-imaging of an E5.5 mouse embryo using incubator-type biaxial light-sheet microscopy.Life science alliance · 2025Article
- The viscoelastic properties of Nicotiana tabacum BY-2 suspension cell lines adapted to high osmolarity.BMC plant biology · 2025Article
- Naltrexone blocks alcohol-induced effects on kappa-opioid receptors in the plasma membrane.Translational psychiatry · 2024Article
- Nucleic Acid-Rich Stress Granules Are Not Merely Crowded Condensates: A Quantitative Raman Imaging Study.Analytical chemistry · 2024Article
- Cell stress and phase separation stabilize the monomeric state of pseudoisocyanine chloride employed as a self-assembly crowding sensor.Communications chemistry · 2024Article
- Cytoplasmic fluidization contributes to breaking spore dormancy in fission yeast.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
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Corrections and comments
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Authors and funding
6 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Hyperosmotic stress activates in live cells numerous processes and also promotes intracellular protein/RNA aggregation and phase separation. However, the time course and the extent of these changes remain largely uncharacterized. To investigate dynamic changes in intracellular macromolecular crowding (MMC) induced by hyperosmotic stress in live cells, we used fluorescence lifetime imaging microscopy and fluorescence correlation spectroscopy (FCS) to quantify changes in the local environment by measuring the fluorescence lifetime and the diffusion of the monomeric enhanced green fluorescent protein (eGFP), respectively. Real-time monitoring of eGFP fluorescence lifetime showed that a faster response to environmental changes due to MMC is observed than when measuring the acceptor/donor emission ratio using the MMC-sensitive Förster resonance energy transfer sensor (GimRET). This suggests that eGFP molecular electronic states and/or collision frequency are affected by changes in the immediate surroundings due to MMC without requiring conformational changes as is the case for the GimRET sensor. Furthermore, eGFP diffusion assessed by FCS indicated higher intracellular viscosity due to increased MMC during hyperosmotic stress. Our findings reveal that changes in eGFP fluorescence lifetime and diffusion are early indicators of elevated intracellular MMC. Our approach can therefore be used to reveal in live cells short-lived transient states through which MMC builds over time, which could not be observed when measuring changes in other physical properties that occur at slower time scales.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.