Intracellular uranium distribution: Comparison of cryogenic fixation versus chemical fixation methods for SIMS analysis

Suhard, David; Tessier, Christine; Manens, Line; Rebière, François; Tack, Karine; Agarande, Michelle; Guéguen, Yann

doi:10.1002/jemt.23047

Cited by 10 publications

(7 citation statements)

References 46 publications

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“…The intracellular dynamics of uranium agreed with the results of rat experiments, where highly concentrated uranium with 50-fold above the mean renal uranium concentration was observed near the nuclei of the S3 segments of the proximal tubule at the early phase of toxicity [17]. In addition, uranium detection has been reported in the perinuclear regions with nerve-derived cells [32], while uranium nuclear translocation in the early phase of exposure has been demonstrated with the liver and kidney cells [33,34]. Uranium colocalization with phosphorus and potassium was observed at the early stages of toxicity, even though uranium compounds precipitate in the cytoplasmic compartment in the form of uranyl phosphate needles after exposure to toxic concentrations (400-2000 μM) of uranium [35][36][37].…”

Section: Discussionsupporting

confidence: 85%

Single-Cell Imaging for Studies of Renal Uranium Transport and Intracellular Behavior

et al. 2021

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Nephrotoxicity is the primary health effect of uranium exposure. However, the renal transport and intracellular behavior of uranium remains to be clearly elucidated. In the present study, the intracellular uranium distribution was examined with the cell lines derived from the S3 segment of mouse renal proximal tubules, which is a toxic target site of uranium, using microbeam-based elemental analysis. Uranium exposure at 100 μM for 24 h (non-toxic phase) was performed in S3 cells. Two types of measurement specimens, including those that are adhesive cell specimens and cryosection specimens, were examined for the positional relationship of the intracellular localization of uranium. Based on the combined results of single-cell imaging from the two types of cell specimens, uranium was distributed inside the cell and localized in the cytoplasm near the cell nucleus. In some cells, uranium was colocalized with phosphorus and potassium. The amount of uranium accumulated in S3 cells was estimated using thin section-standards. The mean uranium content of three adhesive cells was hundreds of femtogram per cell. Thus, we believe that single-cell imaging would be useful for studies on renal uranium transportation and cellular behavior.

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Section: Discussionsupporting

confidence: 85%

Single-Cell Imaging for Studies of Renal Uranium Transport and Intracellular Behavior

et al. 2021

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“…In this study, kidneys were kept in PFA (24 h) and OCT-embedded prior to analysis. The authors are aware that this sample preparation might distort the initial metal concentrations but it poorly alters its distribution [59]. However, the experimental procedure was rigorously applied to minimize the experimental bias.…”

Section: Uranium Exposure and Biological Sample Preparationmentioning

confidence: 99%

A novel calibration strategy based on internal standard–spiked gelatine for quantitative bio-imaging by LA-ICP-MS: application to renal localization and quantification of uranium

et al. 2020

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Mass spectrometry imaging (MSI) using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) has been employed for the elemental bio-distribution and quantification of uranium (U) in histological tissue sections of rodent kidneys. Kidneys were immediately immersed into 4% paraformaldehyde (PFA) solution for 24 h, Tissue-Tek O.C.T. Compound embedded and stored at − 80°C until cutting in a cryostat, and mounted in gel-covered glass slides. In order to assure complete ablation of sample, sample preparation and laser conditions were carefully optimized. In this work, a new analytical methodology is presented for performing quantitative laser ablation analyses based on internal standard (thulium, Tm)-spiked gelatine (10% m/v) for correction of matrix effects, lack of tissue homogeneity, and instrumental drift. In parallel, matrix-matched laboratory standards, dosed at different concentrations of U, were prepared from a pool of rat kidneys. The quantitative images of cryosections revealed heterogeneous distribution of uranium within the renal tissue, because the cortical concentration was up to 120fold higher than the medullary concentration.

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“…Both SIMS technology, and differential centrifugation associated with inductively coupled plasma mass spectrometry measurements, show that U enters cells in culture very rapidly (less than 15 min), probably through the diffusion of soluble forms [73][74][75]. At subtoxic concentrations (<100 µM, i.e., <50 µg/g), soluble U localizes mainly in nuclei, before it precipitates around or inside cells.…”

Section: Cellular Distribution In the Kidneymentioning

confidence: 99%

Review of Knowledge of Uranium-Induced Kidney Toxicity for the Development of an Adverse Outcome Pathway to Renal Impairment

Guéguen

Frerejacques

2022

IJMS

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An adverse outcome pathway (AOP) is a conceptual construct of causally and sequentially linked events, which occur during exposure to stressors, with an adverse outcome relevant to risk assessment. The development of an AOP is a means of identifying knowledge gaps in order to prioritize research assessing the health risks associated with exposure to physical or chemical stressors. In this paper, a review of knowledge was proposed, examining experimental and epidemiological data, in order to identify relevant key events and potential key event relationships in an AOP for renal impairment, relevant to stressors such as uranium (U). Other stressors may promote similar pathways, and this review is a necessary step to compare and combine knowledge reported for nephrotoxicants. U metal ions are filtered through the glomerular membrane of the kidneys, then concentrate in the cortical and juxtaglomerular areas, and bind to the brush border membrane of the proximal convoluted tubules. U uptake by epithelial cells occurs through endocytosis and the sodium-dependent phosphate co-transporter (NaPi-IIa). The identified key events start with the inhibition of the mitochondria electron transfer chain and the collapse of mitochondrial membrane potential, due to cytochrome b5/cytochrome c disruption. In the nucleus, U directly interacts with negatively charged DNA phosphate, thereby inducing an adduct formation, and possibly DNA strand breaks or cross-links. U also compromises DNA repair by inhibiting zing finger proteins. Thereafter, U triggers the Nrf2, NF-κB, or endoplasmic reticulum stress pathways. The resulting cellular key events include oxidative stress, DNA strand breaks and chromosomal aberrations, apoptosis, and pro-inflammatory effects. Finally, the main adverse outcome is tubular damage of the S2 and S3 segments of the kidneys, leading to tubular cell death, and then kidney failure. The attribution of renal carcinogenesis due to U is controversial, and specific experimental or epidemiological studies must be conducted. A tentative construction of an AOP for uranium-induced kidney toxicity and failure was proposed.

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Intracellular uranium distribution: Comparison of cryogenic fixation versus chemical fixation methods for SIMS analysis

Cited by 10 publications

References 46 publications

Single-Cell Imaging for Studies of Renal Uranium Transport and Intracellular Behavior

Single-Cell Imaging for Studies of Renal Uranium Transport and Intracellular Behavior

A novel calibration strategy based on internal standard–spiked gelatine for quantitative bio-imaging by LA-ICP-MS: application to renal localization and quantification of uranium

Review of Knowledge of Uranium-Induced Kidney Toxicity for the Development of an Adverse Outcome Pathway to Renal Impairment

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