Spatial Distribution of Intracellular Ion Concentrations in Aggregate‐Forming HeLa Cells Analyzed by μ‐XRF Imaging

Protein aggregation is a hallmark of several severe neurodegenerative disorders such as Huntington's, Parkinson's, or Alzheimer's disease. Metal ions play a profound role in protein aggregation and altered metal‐ion homeostasis is associated with disease progression. Here we utilize μ‐X‐ray fluorescence imaging in combination with rapid freezing to resolve the elemental distribution of phosphorus, sulfur, potassium, and zinc in huntingtin exon‐1‐mYFP expressing HeLa cells. Using quantitative XRF analysis, we find a threefold increase in zinc and a 10‐fold enrichment of potassium that can be attributed to cellular stress response. While the averaged intracellular ion areal masses are significantly different in aggregate‐containing cells, a local intracellular analysis shows no different ion content at the location of intracellular inclusion bodies. The results are compared to corresponding experiments on HeLa cells forming pseudoisocyanine chloride aggregates. As those show similar results, changes in ion concentrations are not exclusively linked to huntingtin exon‐1 amyloid formation. Quantitative μ‐X‐ray fluorescence imaging was used in combination with rapid freezing to study the elemental distribution in huntingtin‐expressing HeLa cells. A threefold increase in zinc and a 10‐fold enrichment of potassium was found. Corresponding experiments on HeLa cells forming pseudoisocyanine chloride aggregates showed similar results. Both these effects can be attributed to a cellular stress response.