title: Changes to cellular water and element content induced by nucleolar stress: investigation by a cryo-correlative nano-imaging approach.
abstract: The cell is a crowded volume, with estimated mean mass percentage of macromolecules and of water ranging from 7.5 to 45 and 55 to 92.5 %, respectively. However, the concentrations of macromolecules and water at the nanoscale within the various cell compartments are unknown. We recently developed a new approach, correlative cryo-analytical scanning transmission electron microscopy, for mapping the quantity of water within compartments previously shown to display GFP-tagged protein fluorescence on the same ultrathin cryosection. Using energy-dispersive X-ray spectrometry (EDXS), we then identified various elements (C, N, O, P, S, K, Cl, Mg) in these compartments and quantified them in mmol/l. Here, we used this new approach to quantify water and elements in the cytosol, mitochondria, condensed chromatin, nucleoplasm, and nucleolar components of control and stressed cancerous cells. The water content of the control cells was between 60 and 83 % (in the mitochondria and nucleolar fibrillar centers, respectively). Potassium was present at concentrations of 128-462 mmol/l in nucleolar fibrillar centers and condensed chromatin, respectively. The induction of nucleolar stress by treatment with a low dose of actinomycin-D to inhibit rRNA synthesis resulted in both an increase in water content and a decrease in the elements content in all cell compartments. We generated a nanoscale map of water and elements within the cell compartments, providing insight into their changes induced by nucleolar stress.
authors: Laboratoire de Recherche en Nanosciences, Université de Reims Champagne Ardenne, Reims, France.
authors: Frédérique
authors: F
authors: Nolin
authors: Jean
authors: J
authors: Michel
authors: Laurence
authors: L
authors: Wortham
authors: Pavel
authors: P
authors: Tchelidze
authors: Gérard
authors: G
authors: Balossier
authors: Vincent
authors: V
authors: Banchet
authors: Hélène
authors: H
authors: Bobichon
authors: Nathalie
authors: N
authors: Lalun
authors: Christine
authors: C
authors: Terryn
authors: Dominique
authors: D
authors: Ploton
mesh_terms: D002467:Cell Nucleus / Q000737:chemistry* / Q000502:physiology; D002843:Chromatin / Q000737:chemistry; D020285:Cryoelectron Microscopy / Q000379:methods; D016616:Cryoultramicrotomy; D003600:Cytosol / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D042541:Intracellular Space / Q000737:chemistry*; D046911:Macromolecular Substances / Q000737:chemistry; D008855:Microscopy, Electron, Scanning / Q000379:methods; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008928:Mitochondria / Q000737:chemistry; D036103:Nanotechnology; D013052:Spectrometry, X-Ray Emission; D013312:Stress, Physiological; D014867:Water / Q000032:analysis*
chemical_list: D002843:Chromatin; D046911:Macromolecular Substances; D014867:Water
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