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Increased Hippocampal Expression of the Divalent Metal Transporter 1 (DMT1) mRNA Variants 1B and +IRE and DMT1 Protein After NMDA-Receptor Stimulation or Spatial Memory Training

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An Erratum to this article was published on 23 September 2009

Abstract

Iron is essential for crucial neuronal functions but is also highly toxic in excess. Neurons acquire iron through transferrin receptor-mediated endocytosis and via the divalent metal transporter 1 (DMT1). The N-terminus (1A, 1B) and C-terminus (+IRE, −IRE) splice variants of DMT1 originate four protein isoforms, all of which supply iron to cells. Diverse physiological or pathological conditions induce differential DMT1 variant expression, which are cell-type dependent. Hence, it becomes relevant to ascertain if activation of neuronal plasticity processes that require functional N-methyl d-aspartate (NMDA) receptors, including in vitro stimulation of NMDA receptor-mediated signaling and spatial memory training, selectively modify DMT1 variant expression. Here, we report for the first time that brief (5 min) exposure of primary hippocampal cultures to NMDA (50 μM) increased 24 h later the expression of DMT1-1B and DMT1+IRE, but not of DMT1−IRE mRNA. In contrast, endogenous DMT1 mRNA levels remained unaffected following 6 h incubation with brain-derived nerve factor. NMDA (25–50 μM) also enhanced DMT1 protein expression 24–48 h later; this enhancement was abolished by the transcription inhibitor actinomycin D and by the NMDA receptor antagonist MK-801, implicating NMDA receptors in de novo DMT1 expression. Additionally, spatial memory training enhanced DMT1-1B and DMT1+IRE expression and increased DMT1 protein content in rat hippocampus, where the exon1A variant was not found. These results suggest that NMDA receptor-dependent plasticity processes stimulate expression of the iron transporter DMT1-1B+IRE isoform, which presumably plays a significant role in hippocampal spatial memory formation.

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Notes

  1. Haeger P., Muñoz P., Carrasco M. A., Núñez, M. T. & Hidalgo C. Increased RyR and DMT1 expression in rat hippocampus after spatial memory training, 38th annual meeting of the Society for Neuroscience, Washington, USA, November 15–19, 2008.

Abbreviations

ACSF:

Artificial cerebrospinal fluid

Act D:

Actinomycin D

BDNF:

Brain-derived neurotrophic factor

DMT1:

Divalent metal transporter 1

GFAP:

Glial fibrillary acidic protein

IRE:

Iron response element

IRP:

Iron regulatory protein

MAP2:

Microtubule-associated protein 2

MWM:

Morris Water Maze

NF-Y:

Nuclear factor Y

NFκB:

Nuclear factor kappa B

NMDA:

N-Methyl d-aspartate

PBS:

Phosphate buffered saline

ROS:

Reactive oxygen species

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Acknowledgments

This work was supported by Fondap-CEMC 15010006, Fondecyt postdoctoral and doctoral fellowship Grants 3070035 and 24080073, respectively, and by Millennium Scientific Initiative Grant ICM-P05-001-F. The authors thank the skillful professional assistance of N. Leal and P. Fernández and gratefully acknowledge Drs. M.A. Carrasco and K. Gysling for their insightful comments and support, and Dr. N. Inestrosa for kindly providing access to a Morris water maze system.

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Authors and Affiliations

  1. Centro FONDAP de Estudios Moleculares de la Célula, Facultad de Medicina, Universidad de Chile, Independencia 1027, Santiago, Chile

    Paola Haeger, Álvaro Álvarez, Nancy Leal, Tatiana Adasme & Cecilia Hidalgo

  2. Departamento de Biología, Facultad de Ciencias and Institute for Cell Dynamics and Biotechnology, Universidad de Chile, Santiago, Chile

    Marco Tulio Núñez

  3. Programa de Biología Celular y Molecular, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago, Chile

    Cecilia Hidalgo

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  1. Paola Haeger
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  2. Álvaro Álvarez
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Correspondence to Paola Haeger.

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An erratum to this article can be found at http://dx.doi.org/10.1007/s12640-009-9107-0

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Supplementary Fig. 1

Hippocampal primary cultures are enriched in neurons. a Hippocampal cultures were visualized by differential interference contrast (DIC) microscopy (left) or inmunoreacted with MAP2 antibody as neuronal marker (right) and analyzed by confocal microscopy. b Confocal analysis of co-immunofluorescence of β-Tubulin III (red) as neuronal marker and GFAP (green) as glial cell marker, showing neuronal cell enrichment in the culture. Calibration bar, 20 μm (TIFF 3220 kb)

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Haeger, P., Álvarez, Á., Leal, N. et al. Increased Hippocampal Expression of the Divalent Metal Transporter 1 (DMT1) mRNA Variants 1B and +IRE and DMT1 Protein After NMDA-Receptor Stimulation or Spatial Memory Training. Neurotox Res 17, 238–247 (2010). https://doi.org/10.1007/s12640-009-9096-z

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