Highly complex analytical methods with different accuracies of measurement, reproducibilities and ease of analyses are currently being used to quantify metals in cellular media and tissue samples. In this review, the analytical methods commonly used for iron, copper and zinc quantification in mammalian cells are presented and discussed. Herein, we present a literature survey of the most commonly found concentrations of these metals in various mammalian cells in culture and tissues. The aim of this review is to help researchers in metallomic-related areas identify the method that best suits their needs for the accurate quantification of these metals in cells. This accuracy goes beyond simple knowledge of the limit of detection of each technique and needs to be evaluated through comparisons with similar previous studies.
The PrP(C) protein, which is especially present in the cellular membrane of nervous system cells, has been extensively studied for its controversial antioxidant activity. In this study, we elucidated the free radical scavenger activity of purified murine PrP(C) in solution and its participation as a cell protector in astrocytes that were subjected to treatment with an oxidant. In vitro and using an EPR spin-trapping technique, we observed that PrP(C) decreased the oxidation of the DMPO trap in a Fenton reaction system (Cu(2+)/ascorbate/H(2)O(2)), which was demonstrated by approximately 70% less DMPO/OH(). In cultured PrP(C)-knockout astrocytes from mice, the absence of PrP(C) caused an increase in intracellular ROS (reactive oxygen species) generation during the first 3h of H(2)O(2) treatment. This rapid increase in ROS disrupted the cell cycle in the PrP(C)-knockout astrocytes, which increased the population of cells in the sub-G1 phase when compared with cultured wild-type astrocytes. We conclude that PrP(C) in solution acts as a radical scavenger, and in astrocytes, it is essential for protection from oxidative stress caused by an external chemical agent, which is a likely condition in human neurodegenerative CNS disorders and pathological conditions such as ischemia.
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