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After cooling, the glass coverslips were broken off, and the samples were cut using an ultramicrotome (EM UC6, Leica Microsystems, Wetzlar, Germany) into thin sections of ~200 nm, and then placed on Silicon wafers (Siegert Wafer GmbH, Aachen, Germany)

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In other mammals, such as rodents, it has been reported that the SNc of aged rats shows an important decrease in TH protein content, accompanied by a significant reduction in dopamine levels

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HCD fragmentation was applied with 27% collision energy and resulting fragments were detected using the rapid scan rate in the Orbitrap