Localization and absolute quantification of dopamine in discrete intravesicular compartments using nanoSIMS imaging

Rabasco, StefaniaGothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology (författare)

The absolute concentration and the compartmentalization of analytes in cells and organelles are crucial parameters in the development of drugs and drug delivery systems, as well as in the fundamental understanding of many cellular processes. Nanoscale secondary ion mass spectrometry (NanoSIMS) imaging is a powerful technique which allows subcellular localization of chemical species with high spatial and mass resolution, and high sensitivity. In this study, we combined NanoSIMS imaging with spatial oversampling with transmission electron microscopy (TEM) imaging to discern the compartments (dense core and halo) of large dense core vesicles in a model cell line used to study exocytosis, and to localize13C dopamine enrichment following 4–6 h of 150 μM13C L-3,4-dihydroxyphenylalanine (L-DOPA) incubation. In addition, the absolute concentrations of13C dopamine in distinct vesicle domains as well as in entire single vesicles were quantified and validated by comparison to electrochemical data. We found concentrations of 87.5 mM, 16.0 mM and 39.5 mM for the dense core, halo and the whole vesicle, respectively. This approach adds to the potential of using combined TEM and NanoSIMS imaging to perform absolute quantification and directly measure the individual contents of nanometer-scale organelles.

Nguyen, Tho D. K.Gothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology(Swepub:gu)xngduc (författare)
Gu, Chaoyi,1992Gothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology(Swepub:gu)xgucha (författare)
Kurczy, Michael E. (författare)
Phan, Nhu TN,1981Gothenburg University,Göteborgs universitet,National Center for Imaging Mass Spectrometry,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology(Swepub:gu)xphant (författare)
Ewing, Andrew G,1957Gothenburg University,Göteborgs universitet,National Center for Imaging Mass Spectrometry,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology(Swepub:gu)xewian (författare)
Göteborgs universitetInstitutionen för kemi och molekylärbiologi (creator_code:org_t)

Ingår i:International Journal of Molecular Sciences: MDPI AG23:11661-65961422-0067

Av författaren/redakt...: Rabasco, Stefani ...; Nguyen, Tho D. K ...; Gu, Chaoyi, 1992; Kurczy, Michael ...; Phan, Nhu TN, 19 ...; Ewing, Andrew G, ...

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