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Cellular and sub-cellular Cu isotope fractionation in the human neuroblastoma SH-SY5Y cell line: proliferating versus neuron-like cells

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Abstract

Cu isotope fractionation was investigated in the human neuroblastoma SH-SY5Y cell line, in a proliferating/tumor phase (undifferentiated cells), and in a differentiated state (neuron-like cells), induced using retinoic acid (RA). The SH-SY5Y cell line displays genetic aberrations due to its cancerous origin, but differentiation drives the cell line towards phenotypes suitable for the research of neurological diseases (e.g., Alzheimer’s disease or Parkinson’s disease). Cellular Cu distribution was first explored by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) imaging and, subsequently, Cu isotopic analysis was performed at cellular and sub-cellular levels via multi-collector inductively coupled plasma-mass spectrometry (MC-ICP-MS). The SH-SY5Y cells showed a re-distribution of intracellular Cu upon RA differentiation. Both undifferentiated and differentiated cells became systematically enriched in the light 63Cu isotope with increasing intracellular Cu content. Differentiated neuron-like SH-SY5Y cells showed a heavier Cu isotopic composition (+ 0.3‰) than did the undifferentiated proliferating cells when exposed to Cu for 24 h. However, after a longer exposure time (72 h), no difference was observed between both cellular phenotypes. Mitochondrial fractions were enriched in the light 63Cu isotope, compared to whole cells, for both undifferentiated and differentiated cells (no significant difference). The Cu isotopic composition of the remaining cell lysates was heavier than that of the whole cells and + 0.2‰ heavier in the differentiated cells than in the undifferentiated cells. These results indicate that neuronal differentiation affects the Cu isotope fractionation accompanying Cu uptake in the cells, but this effect does not seem to be associated with the mitochondrial Cu pathway. Cu isotope fractionation can be an interesting tool for studying Cu metabolism at a (sub)-cellular level in functional neurons.

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Acknowledgements

Marta Costas-Rodríguez thanks FWO-Vlaanderen for her postdoctoral grant. Legna Colina-Vegas acknowledges the São Paulo Research Foundation (Fapesp grant #2017/23254-9). The authors would like to acknowledge Teledyne CETAC Technologies Inc. for logistic support.

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Authors and Affiliations

  1. Department of Chemistry, Atomic & Mass Spectrometry – A&MS Research Unit, Ghent University, Campus Sterre, Krijgslaan 281-S12, 9000, Ghent, Belgium

    Marta Costas-Rodríguez, Legna Colina-Vegas, Nikolay Solovyev & Frank Vanhaecke

  2. Departamento de Química, Universidade Federal de São Carlos – UFSCar, Rodovia Washington Luís KM 235, CP 676, São Carlos, SP, 13565-905, Brazil

    Legna Colina-Vegas

  3. Department of Human Structure and Repair, Laboratory of Experimental Cancer Research, Ghent University, Corneel Heymanslaan 10, 9000, Ghent, Belgium

    Olivier De Wever

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  1. Marta Costas-Rodríguez
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Correspondence to Marta Costas-Rodríguez.

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Published in the topical collection Young Investigators in (Bio-)Analytical Chemistry with guest editors Erin Baker, Kerstin Leopold, Francesco Ricci, and Wei Wang.

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Costas-Rodríguez, M., Colina-Vegas, L., Solovyev, N. et al. Cellular and sub-cellular Cu isotope fractionation in the human neuroblastoma SH-SY5Y cell line: proliferating versus neuron-like cells. Anal Bioanal Chem 411, 4963–4971 (2019). https://doi.org/10.1007/s00216-019-01871-6

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  • DOI: https://doi.org/10.1007/s00216-019-01871-6

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