Because ferroportin (Fpn) is the only known mammalian cellular iron exporter, understanding its localization and regulation within the retina would shed light on the direction of retinal iron flux. The hormone hepcidin may regulate retinal Fpn, as it triggers Fpn degradation in the gut. Immunofluorescence was used to label Fpn in retinas of mice with 4 different genotypes (wild type; Fpn C326S, a hepcidin-resistant Fpn; hepcidin knockout; and ceruloplasmin/hephaestin double knockout). No significant difference in Fpn levels was observed in these retinas. Fpn localized to the abluminal side of the outer plexiform vascular endothelial cells, Müller glia cells, and the basolateral side of the retinal pigment epithelium. Adenoassociated virus (AAV)-hepcidin was injected into the eyes of hepcidin knockout mice, while AAV-lacZ was injected into the contralateral eyes as a control. AAV-hepcidin injected eyes had increased ferritin immunolabeling in retinal vascular endothelial cells. Fpn C326S mice had systemic iron overload compared to wild type and had the fastest retinal iron accumulation of any hereditary model studied to date. The results suggest that physiologic hepcidin levels are insufficient to alter Fpn levels within the retinal pigment epithelium and Müller cells, but may limit iron transport into the retina from vascular endothelial cells.-Theurl, M., Song, D., Clark, E., Sterling, J., Grieco, S., Altamura, S., Galy, B., Hentze, M., Muckenthaler, M. U., Dunaief, J. L. Mice with hepcidin-resistant ferroportin accumulate iron in the retina. FASEB J. 30, 813-823 (2016). www.fasebj.org Key Words: retinal pigment epithelium • vascular endothelium • iron transport • blood-brain barrier Iron, while essential for living organisms, must be tightly regulated as it can catalyze the formation of reactive oxygen species. Within the retina, labile iron has been shown to cause degeneration (1). Age-related macular degeneration (AMD), the leading cause of irreversible vision loss in individuals over 50 yr old (2), is associated with increased retinal iron (3). We have previously studied mice lacking the 2 ferroxidases ceruloplasmin (Cp) and hephaestin (Heph), which leads to intracellular iron entrapment. These double-knockout (DKO) mice develop retinal iron overload, causing retinal degeneration with some histologic and molecular features of human AMD (4). Retinal degeneration in the ceruloplasmin/hephaestin DKO (Cp/Heph DKO) mice can be ameliorated by systemic iron chelation therapy (5). A role for iron in AMD is further supported by the observation that patients with aceruloplasminemia, an autosomal recessive disease with retinal iron accumulation, can develop early-onset macular degeneration (6, 7).Ferroportin (Fpn), a transmembrane protein that is the only known cellular iron exporter (8), and hepcidin, an iron regulatory hormone, are 2 key contributors to retinal iron homeostasis. In enterocytes and macrophages, hepcidin binds to Fpn, triggering its internalization and degradation (9). Knockout (KO) of hepcidi...
Mutations in the RP1 gene can cause retinitis pigmentosa. We identified a spontaneous L66P mutation caused by two adjacent point mutations in the Rp1 gene in a colony of C57BL/6J mice. Mice homozygous for the L66P mutation exhibited slow, progressive photoreceptor degeneration throughout their lifespan. Optical coherence tomography imaging found abnormal photoreceptor reflectivity at 1 month of age. Histology found shortening and disorganization of the photoreceptor inner and outer segments and progressive thinning of the outer nuclear layer. Electroretinogram a- and b-wave amplitudes were decreased with age. Western blot analysis found that the quantity and size of the mutated retinitis pigmentosa 1 (RP1) protein were normal. However, immunohistochemistry found that the mutant Rp1 protein partially mislocalized to the transition zone of the shortened axonemes. This mutation disrupted colocalization with cytoplasmic microtubules in vitro. In conclusion, the L66P mutation in the first doublecortin domain of the Rp1 gene impairs Rp1 protein localization and function, leading to abnormalities in photoreceptor outer segment structure and progressive photoreceptor degeneration. This is the first missense mutation in Rp1 shown to cause retinal degeneration. It provides a unique, slowly progressive photoreceptor degeneration model that mirrors the slow degeneration kinetics in most patients with retinitis pigmentosa.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.