Coherent Raman microscopy visualizes ongoing cellular senescence through amide I peak shifts originating from β sheets in disordered nucleolar proteins.

Publisher: Nature Publishing Group Country of Publication: England NLM ID: 101563288 Publication Model: Electronic Cited Medium: Internet ISSN: 2045-2322 (Electronic) Linking ISSN: 20452322 NLM ISO Abbreviation: Sci Rep Subsets: MEDLINE

Original Publication: London : Nature Publishing Group, copyright 2011-

Cellular Senescence* ; Spectrum Analysis, Raman*/methods ; Cell Nucleolus*/metabolism; Humans ; Protein Conformation, beta-Strand ; Amides/chemistry ; Nuclear Proteins/metabolism

Cellular senescence occurs through the accumulation of many kinds of stresses. Senescent cells in tissues also cause various age-related disorders. Therefore, detecting them without labeling is beneficial for medical research and developing diagnostic methods. However, existing biomarkers have limitations of requiring fixation and labeling, or their molecular backgrounds are uncertain. Coherent anti-Stokes Raman scattering (CARS) spectroscopic imaging is a novel option because it can assess and visualize molecular structures based on their molecular fingerprint. Here, we present a new label-free method to visualize cellular senescence using CARS imaging in nucleoli. We found the peak of the nucleolar amide I band shifted to a higher wavenumber in binuclear senescent cells, which reflects changes in the protein secondary structure from predominant α-helices to β-sheets originating from amyloid-like aggregates. Following this, we developed a procedure that can visualize the senescent cells by providing the ratios and subtractions of these two components. We also confirmed that the procedure can visualize nucleolar aggregates due to unfolded/misfolded proteins produced by proteasome inhibition. Finally, we found that this method can help visualize the nucleolar defects in naïve cells even before binucleation. Thus, our method is beneficial to evaluate ongoing cellular senescence through label-free imaging of nucleolar defects.
Competing Interests: Declarations Competing interests The authors declare no competing interests.
(© 2024. The Author(s).)

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19K06671 Japan Society for the Promotion of Science; 18H02000 Japan Society for the Promotion of Science; ANR-10-LABX-0074 Sigma-LIM Agence Nationale de la Recherche; JPMJMI22G5 JST-Mirai Program

Keywords: Amide I; Cellular senescence; Coherent anti-stokes Raman scattering (CARS) microscopy; LLPS (liquid-liquid phase separation); Nucleolus; Raman

0 (Amides)
0 (Nuclear Proteins)

Date Created: 20241111 Date Completed: 20241111 Latest Revision: 20241116

20241116

PMC11555345

10.1038/s41598-024-78899-x

39528609