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Receptor‐Mediated Endocytosis of a Manganese Complex of Transferrin into Neuroblastoma (SHSY5Y) Cells in Culture

Abstract: Exposure to manganese compounds often occurs as the result of industrial production or mining. Although manganese appears in traces in animal and human tissue and is essential to certain biological processes, it is also toxic. In humans and animals, toxicity is mainly associated with the nervous system. The mechanism underlying behavioral and biochemical alterations observed after manganese intoxication is not fully understood. We have shown that the manganese present in serum after exposure to manganese oxide is bound to transferrin as trivalent manganic ion. In this study of manganese uptake and storage we used a clone of human neuroblastoma cells (SHSY5Y). These cells differentiate and express catechol‐aminergic properties. Saturation binding analysis of the transferrin‐manganese complex to the cells revealed a single class of binding sites, with an apparent KD of 13 ± 1 nM and a density of 11, 000 ± 2, 000 binding sites per cell. The complex was internalized in a temperature‐dependent way and reached saturation after 2 h when ∼2% of the added manganese had been internalized. About 80% of the internalized manganese was found in ferritin after 24 h of exposure. The results demonstrate that the transferrin receptor on SHSY5Y cells can bind and internalize a manganese‐transferrin complex as efficiently as an iron‐transferrin complex, although a saturation of the manganese uptake was achieved.