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Molecular Neurobiology

, Volume 56, Issue 4, pp 2362–2374 | Cite as

Hepcidin Mediates Transcriptional Changes in Ferroportin mRNA in Differentiated Neuronal-Like PC12 Cells Subjected to Iron Challenge

  • Steinunn Sara Helgudottir
  • Jacek Lichota
  • Annette Burkhart
  • Torben MoosEmail author
Article
  • 179 Downloads

Abstract

Ferroportin is the only known iron exporter, and its regulation seems to be controlled at both transcriptional, post-transcriptional, and post-translational levels. The objective of the current work was to investigate how cellular iron status affects the expression of the ferroportin gene Fpn under the influence of hepcidin, known to post-translational lower the available ferroportin protein. Nerve growth factor-beta (NGF-β)-differentiated PC12 cells, used as a model of neuronal cells, were evaluated in terms of their viability and expression of ferroportin after inducing cellular iron overload with ferric ammonium citrate (FAC) or hepcidin, iron deficiency with deferoxamine (DFO), or hepcidin in combination with FAC or DFO. Ferritin mRNA was significantly upregulated following treatment with 20 mM FAC. The viability of the differentiated PC12 cells was significantly reduced after treatment with 30 mM FAC or 1.0 μM hepcidin, but when combining FAC and hepcidin treatment, the cells remained unaffected. The expression of Fpn was concurrently upregulated after treatment with FAC in combination with hepcidin. Fifty millimolar DFO also increased Fpn. Together, these data point towards a transcriptional induction of Fpn in response to changes in cellular iron levels. Epigenetic regulation of Fpn may also occur as changes in genes associated with epigenetic regulation of Fpn were demonstrated.

Keywords

Ferroportin Iron Neurodegeneration Hepcidin Epigenetic 

Abbreviations

ARE

Antioxidant response element

BSA

Bovine serum albumin

Cy5

Cyanine-5

DAPI

4′,6-Diamidino-2-phenylindole dihydrochloride

DFO

Deferoxamine

Dmt1

Divalent metal transporter 1

DIV

Days in vitro

FAC

Ferric ammonium iron (III) citrate

FCS

Fetal calf serum

Fe2+

Ferrous iron

Fe3+

Ferric iron

Fpn

Ferroportin gene expression

Ftl

Ferritin light chain

Fth

Ferritin heavy chain

Hdac1

Histone deacetylase 1

IRE

Iron-responsive element

IRP

Iron regulatory protein

NGF-β

Nerve growth factor beta1

PBS

Phosphate-buffered saline

PFA

Paraformaldehyde

Phf8

PHD finger protein 8

RNS

Reactive nitrogen species

ROS

Reactive oxygen species

SEM

Standard error of the mean

Tet1

Ten-eleven translocation methylcytosine dioxygenase 1

Tubb4b

Class IV β-tubulin

UTR

Untranslated region

Notes

Acknowledgments

We thank Poul Henning Jensen for providing PC12 cells and Merete Fredsgaard and Hanne Krone Nielsen, Aalborg University, Denmark, for the excellent technical assistance. We thank Assistant Professor Maj Schneider Thomsen, Aalborg University, Denmark, for the illustrative work.

Funding Information

The present work has been supported by The Danish Multiple Sclerosis Society, “Fonden til Lægevidenskabens Fremme,” Augustinus fonden, and the “Åse og Ejner Danielsens Fond.”

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interests.

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Laboratory of Neurobiology, Biomedicine Group, Department of Health Science and TechnologyAalborg UniversityAalborg EastDenmark

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