Detection and quantitation of iron in ferritin, transferrin and labile iron pool (LIP) in cardiomyocytes using 55Fe and storage phosphorimaging
Jazyk angličtina Země Nizozemsko Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
30279145
DOI
10.1016/j.bbagen.2018.09.005
PII: S0304-4165(18)30293-9
Knihovny.cz E-zdroje
- Klíčová slova
- Cardiomyocytes, LIP, Phosphorimaging, Radioactive iron, Spin micro columns,
- MeSH
- biologický transport MeSH
- chelátory chemie MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- ferritiny chemie metabolismus MeSH
- gelová chromatografie MeSH
- homeostáza MeSH
- kardiomyocyty metabolismus MeSH
- kultivované buňky MeSH
- limita detekce MeSH
- potkani Wistar MeSH
- radioizotopy železa metabolismus MeSH
- transferin chemie metabolismus MeSH
- železo chemie metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- chelátory MeSH
- ferritiny MeSH
- radioizotopy železa MeSH
- transferin MeSH
- železo MeSH
Dysregulated iron metabolism has a detrimental effect on cardiac function. The importance of iron homeostasis in cardiac health and disease warrants detailed studies of cardiomyocyte iron uptake, utilization and recycling at the molecular level. In this study, we have performed metabolic labeling of primary cultures of neonatal rat cardiomyocytes with radioactive iron coupled with separation of labeled iron-containing molecules by native electrophoresis followed by detection and quantification of incorporated radioiron by storage phosphorimaging. For the radiolabeling we used a safe and convenient beta emitter 55Fe which enabled sensitive and simultaneous detection and quantitation of iron in cardiomyocyte ferritin, transferrin and the labile iron pool (LIP). The LIP is believed to represent potentially dangerous redox-active iron bound to uncharacterized molecules. Using size-exclusion chromatography spin micro columns, we demonstrate that iron in the LIP is bound to high molecular weight molecule(s) (≥5000 Da) in the neonatal cardiomyocytes.
Department of Cardiology Institute for Clinical and Experimental Medicine Prague Czech Republic
Institute of Hematology and Blood Transfusion Prague Czech Republic
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