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. 2013 Oct 7;74(6):848–861. doi: 10.1002/ana.23974

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© 2013 The Authors. Annals of Neurology published by Wiley Periodicals, Inc. on behalf of the American Neurological Association.

This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Iron and the expression of iron-related proteins in the white matter (WM) of controls and multiple sclerosis (MS) patients. (A) Luxol fast blue–periodic acid Schiff myelin staining (blue) and (B) total nonheme iron (brown) of consecutive double-hemispheric slides of MS Subject 25. High iron load is visible in the basal ganglia and the leukocortical boundary. Multiple demyelinated WM lesions, which harbor less iron than the normal-appearing WM (NAWM), are indicated by arrows. (C–Q) Presence of total nonheme iron and iron-related proteins in the normal WM (NWM) of controls (left panel of images; C, F, I, L, O), in the NAWM of MS patients (middle panel; D, G, J, M, P), and in the periplaque WM (PPWM) of MS patients close to the edge of an active lesion (right panel; E, H, K, N, Q). (C–E) In the NWM (C), iron is mainly present in myelin and cells with oligodendrocytic morphology (inset in C) and is reduced in the NAWM of MS patients (D). Inset in D shows small round cells indicative of oligodendrocytes and a larger cell indicative of activated microglia morphology. Close to active plaques, iron is largely lost from oligodendrocytes and myelin, but is present in activated microglia cells (E). (F–H) Immunohistochemistry for ferritin (brown) shows a similar cellular distribution as iron; however, more cells are detected with ferritin immunohistochemistry than with total nonheme iron staining. (I–N) Hephaestin (blue) is expressed in the NWM in oligodendrocytes (I; oligodendrocyte marker TPPP/p25 = red in I–K) and in astrocytes (L; astrocyte marker glial fibrillary acidic protein = red in L–N). In the MS NAWM, both hephaestin⁺ oligodendrocytes (J) and astrocytes (M) are reduced. In the PPWM close to active lesions, hephaestin is upregulated in relation to the NAWM in oligodendrocytes (K), but not in astrocytes (N). (O–Q) Ceruloplasmin (brown) is mainly observed in astrocytes. Its expression is consistently low in the NWM (O), elevated in the NAWM of some MS patients (P), and consistently strong at the edge of active lesions (Q). AG = astroglia; OG = oligodendroglia; scale bars = 100μm; inset scale bars = 10μm (C, I–N) or 25μm (D–H).