Oligodendrocyte progenitor cells internalize ferritin via clathrin-dependent receptor mediated endocytosis.
We previously demonstrated ferritin binding is specific to white matter in mouse and human brain tissue and is not found within Multiple Sclerotic plaques. These results suggest that ferritin receptors are selectively expressed on oligodendrocytes. The present studies were designed to test the hypothesis that oligodendrocyte progenitor cells selectively bind ferritin and internalize it by methods consistent with receptor-mediated endocytosis. Using a cell culture system enriched for oligodendrocyte progenitor cells, we determined, that oligodendrocyte progenitor cells bind ferritin in a saturable and competitive manner with a K(d) of 5 nM and a receptor density of 0.06 fmol bound/20,000 cells. FITC tagged ferritin is internalized by A2B5, O4 or CNPase expressing cells in the culture, but not by GFAP+ cells. The uptake of ferritin into the oligodendrocyte progenitors was inhibited by treating the cells with inhibitors of receptor mediated endocytosis (hypertonic medium, potassium deficient medium, ATP depletion, sulfhydryl reagents). In addition exogenous ferritin decreased iron responsive element/iron regulatory protein binding indicating that the iron within the internalized ferritin is released and contributes to the intracellular iron pool. Given the relatively high amount of iron that can be delivered via ferritin, and the selective distribution of ferritin receptors in the white matter tracts in vivo, we propose that ferritin is a major source of iron for oligodendrocytes.
Hulet
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SW
George M. Leader Family Laboratory for Alzheimer Disease Research, Department of Neuroscience and Anatomy, M.S. Hershey Medical Center, Pennsylvania State University College of Medicine, Hershey 17033, USA.
Heyliger
S O
SO
Powers
S
S
Connor
J R
JR
D000818:Animals; D001667:Binding, Competitive / Q000502:physiology; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D001921:Brain / Q000166:cytology; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D002966:Clathrin / Q000378:metabolism*; D003676:Deferoxamine / Q000494:pharmacology; D004705:Endocytosis / Q000502:physiology*; D005293:Ferritins / Q000493:pharmacokinetics*; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006801:Humans; D007457:Iodine Radioisotopes; D007501:Iron / Q000378:metabolism; D009103:Multiple Sclerosis / Q000378:metabolism; D009186:Myelin Sheath / Q000378:metabolism; D009474:Neurons / Q000166:cytology; D009836:Oligodendroglia / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D011990:Receptors, Transferrin / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D014168:Transferrin / Q000493:pharmacokinetics
D002614:Chelating Agents; D002966:Clathrin; D005456:Fluorescent Dyes; D007457:Iodine Radioisotopes; D011990:Receptors, Transferrin; D014168:Transferrin; D005293:Ferritins; D007501:Iron; D016650:Fluorescein-5-isothiocyanate; D003676:Deferoxamine