Effect of iron overload and iron deficiency on liver hemojuvelin protein.

INTRODUCTION: Hemojuvelin (Hjv) is a key component of the signaling cascade that regulates liver hepcidin (Hamp) expression. The purpose of this study was to determine Hjv protein levels in mice and rats subjected to iron overload and iron deficiency. METHODS: C57BL/6 mice were injected with iron (2...

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Main Authors: Jan Krijt, Jana Frýdlová, Lenka Kukačková, Yuzo Fujikura, Petr Přikryl, Martin Vokurka, Emanuel Nečas
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2012-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC3356351?pdf=render
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author Jan Krijt
Jana Frýdlová
Lenka Kukačková
Yuzo Fujikura
Petr Přikryl
Martin Vokurka
Emanuel Nečas
author_facet Jan Krijt
Jana Frýdlová
Lenka Kukačková
Yuzo Fujikura
Petr Přikryl
Martin Vokurka
Emanuel Nečas
author_sort Jan Krijt
collection DOAJ
description INTRODUCTION: Hemojuvelin (Hjv) is a key component of the signaling cascade that regulates liver hepcidin (Hamp) expression. The purpose of this study was to determine Hjv protein levels in mice and rats subjected to iron overload and iron deficiency. METHODS: C57BL/6 mice were injected with iron (200 mg/kg); iron deficiency was induced by feeding of an iron-deficient diet, or by repeated phlebotomies. Erythropoietin (EPO)-treated mice were administered recombinant EPO at 50 U/mouse. Wistar rats were injected with iron (1200 mg/kg), or fed an iron-deficient diet. Hjv protein was determined by immunoblotting, liver samples from Hjv-/- mice were used as negative controls. Mouse plasma Hjv content was determined by a commercial ELISA kit. RESULTS: Liver crude membrane fraction from both mice and rats displayed a major Hjv-specific band at 35 kDa, and a weaker band of 20 kDa. In mice, the intensity of these bands was not changed following iron injection, repeated bleeding, low iron diet or EPO administration. No change in liver crude membrane Hjv protein was observed in iron-treated or iron-deficient rats. ELISA assay for mouse plasma Hjv did not show significant difference between Hjv+/+ and Hjv-/- mice. Liver Hamp mRNA, Bmp6 mRNA and Id1 mRNA displayed the expected response to iron overload and iron deficiency. EPO treatment decreased Id1 mRNA, suggesting possible participation of the bone morphogenetic protein pathway in EPO-mediated downregulation of Hamp mRNA. DISCUSSION: Since no differences between Hjv protein levels were found following various experimental manipulations of body iron status, the results indicate that, in vivo, substantial changes in Hamp mRNA can occur without noticeable changes of membrane hemojuvelin content. Therefore, modulation of hemojuvelin protein content apparently does not represent the limiting step in the control of Hamp gene expression.
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spelling doaj.art-5427124da8e6428cbf9633afb29c4bcb2022-12-21T18:48:33ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0175e3739110.1371/journal.pone.0037391Effect of iron overload and iron deficiency on liver hemojuvelin protein.Jan KrijtJana FrýdlováLenka KukačkováYuzo FujikuraPetr PřikrylMartin VokurkaEmanuel NečasINTRODUCTION: Hemojuvelin (Hjv) is a key component of the signaling cascade that regulates liver hepcidin (Hamp) expression. The purpose of this study was to determine Hjv protein levels in mice and rats subjected to iron overload and iron deficiency. METHODS: C57BL/6 mice were injected with iron (200 mg/kg); iron deficiency was induced by feeding of an iron-deficient diet, or by repeated phlebotomies. Erythropoietin (EPO)-treated mice were administered recombinant EPO at 50 U/mouse. Wistar rats were injected with iron (1200 mg/kg), or fed an iron-deficient diet. Hjv protein was determined by immunoblotting, liver samples from Hjv-/- mice were used as negative controls. Mouse plasma Hjv content was determined by a commercial ELISA kit. RESULTS: Liver crude membrane fraction from both mice and rats displayed a major Hjv-specific band at 35 kDa, and a weaker band of 20 kDa. In mice, the intensity of these bands was not changed following iron injection, repeated bleeding, low iron diet or EPO administration. No change in liver crude membrane Hjv protein was observed in iron-treated or iron-deficient rats. ELISA assay for mouse plasma Hjv did not show significant difference between Hjv+/+ and Hjv-/- mice. Liver Hamp mRNA, Bmp6 mRNA and Id1 mRNA displayed the expected response to iron overload and iron deficiency. EPO treatment decreased Id1 mRNA, suggesting possible participation of the bone morphogenetic protein pathway in EPO-mediated downregulation of Hamp mRNA. DISCUSSION: Since no differences between Hjv protein levels were found following various experimental manipulations of body iron status, the results indicate that, in vivo, substantial changes in Hamp mRNA can occur without noticeable changes of membrane hemojuvelin content. Therefore, modulation of hemojuvelin protein content apparently does not represent the limiting step in the control of Hamp gene expression.http://europepmc.org/articles/PMC3356351?pdf=render
spellingShingle Jan Krijt
Jana Frýdlová
Lenka Kukačková
Yuzo Fujikura
Petr Přikryl
Martin Vokurka
Emanuel Nečas
Effect of iron overload and iron deficiency on liver hemojuvelin protein.
PLoS ONE
title Effect of iron overload and iron deficiency on liver hemojuvelin protein.
title_full Effect of iron overload and iron deficiency on liver hemojuvelin protein.
title_fullStr Effect of iron overload and iron deficiency on liver hemojuvelin protein.
title_full_unstemmed Effect of iron overload and iron deficiency on liver hemojuvelin protein.
title_short Effect of iron overload and iron deficiency on liver hemojuvelin protein.
title_sort effect of iron overload and iron deficiency on liver hemojuvelin protein
url http://europepmc.org/articles/PMC3356351?pdf=render
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