Cellular iron concentrations directly affect the expression levels of norepinephrine transporter in PC12 cells and rat brain tissue

John L. Beard, Jason A. Wiesinger, Byron Jones

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Neurological development and functioning are adversely affected by iron deficiency in early life. Iron-deficient rats are known to have elevations in extracellular DA and NE, suggesting alterations in reuptake of these monoamines. To explore possible mechanisms by which cellular iron concentrations may alter NE transporter functioning, we utilized NET expressing PC12 cells and iron-deficient rats to explore the relationship between NET protein and mRNA expression patterns and iron concentrations. Treatment of PC12 with the iron chelator, desferrioxamine mesylate (DFO, 50 μM for 24 h), significantly decreased [3H] NE uptake by more than 35% with no apparent change in Km. PC12 cells exposed to increasing concentrations of DFO (25-100 μM) exhibited a dose response decrease in [3H] NE uptake within 24 h (38-73% of control) that paralleled a decrease in cellular NET protein content. Inhibition of protein synthesis with cycloheximide resulted in NET disappearance rates from DFO-treated cells greatly exceeding the rate of loss from control cells. RT-PCR analysis revealed only a modest decrease in NET mRNA levels. Rat brain locus ceruleus and thalamus NET mRNA levels were also only modestly decreased (10-15%) despite a 40% reduction in regional brain iron. In contrast, NET proteins levels in thalamus and locus ceruleus were strongly affected by regional iron deficiency with high correlations with iron concentrations (r > 0.94 and r > 0.80 respectively). The present findings demonstrate that NET protein concentrations and functioning are dramatically reduced with iron deficiency; the modest effect on mRNA levels suggests a stronger influence on NET trafficking and degradation than on protein synthesis.

Original languageEnglish (US)
Pages (from-to)47-58
Number of pages12
JournalBrain Research
Volume1092
Issue number1
DOIs
StatePublished - May 30 2006

Fingerprint

Norepinephrine Plasma Membrane Transport Proteins
PC12 Cells
Iron
Brain
Messenger RNA
Locus Coeruleus
Proteins
Thalamus
Mesylates
Deferoxamine
Cycloheximide
Chelating Agents
Proteolysis

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)
  • Molecular Biology
  • Clinical Neurology
  • Developmental Biology

Cite this

Cellular iron concentrations directly affect the expression levels of norepinephrine transporter in PC12 cells and rat brain tissue. / Beard, John L.; Wiesinger, Jason A.; Jones, Byron.

In: Brain Research, Vol. 1092, No. 1, 30.05.2006, p. 47-58.

Research output: Contribution to journalArticle

@article{728c16b94b8f4df7b44530b9cfd5b05d,
title = "Cellular iron concentrations directly affect the expression levels of norepinephrine transporter in PC12 cells and rat brain tissue",
abstract = "Neurological development and functioning are adversely affected by iron deficiency in early life. Iron-deficient rats are known to have elevations in extracellular DA and NE, suggesting alterations in reuptake of these monoamines. To explore possible mechanisms by which cellular iron concentrations may alter NE transporter functioning, we utilized NET expressing PC12 cells and iron-deficient rats to explore the relationship between NET protein and mRNA expression patterns and iron concentrations. Treatment of PC12 with the iron chelator, desferrioxamine mesylate (DFO, 50 μM for 24 h), significantly decreased [3H] NE uptake by more than 35{\%} with no apparent change in Km. PC12 cells exposed to increasing concentrations of DFO (25-100 μM) exhibited a dose response decrease in [3H] NE uptake within 24 h (38-73{\%} of control) that paralleled a decrease in cellular NET protein content. Inhibition of protein synthesis with cycloheximide resulted in NET disappearance rates from DFO-treated cells greatly exceeding the rate of loss from control cells. RT-PCR analysis revealed only a modest decrease in NET mRNA levels. Rat brain locus ceruleus and thalamus NET mRNA levels were also only modestly decreased (10-15{\%}) despite a 40{\%} reduction in regional brain iron. In contrast, NET proteins levels in thalamus and locus ceruleus were strongly affected by regional iron deficiency with high correlations with iron concentrations (r > 0.94 and r > 0.80 respectively). The present findings demonstrate that NET protein concentrations and functioning are dramatically reduced with iron deficiency; the modest effect on mRNA levels suggests a stronger influence on NET trafficking and degradation than on protein synthesis.",
author = "Beard, {John L.} and Wiesinger, {Jason A.} and Byron Jones",
year = "2006",
month = "5",
day = "30",
doi = "10.1016/j.brainres.2006.03.071",
language = "English (US)",
volume = "1092",
pages = "47--58",
journal = "Brain Research",
issn = "0006-8993",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Cellular iron concentrations directly affect the expression levels of norepinephrine transporter in PC12 cells and rat brain tissue

AU - Beard, John L.

AU - Wiesinger, Jason A.

AU - Jones, Byron

PY - 2006/5/30

Y1 - 2006/5/30

N2 - Neurological development and functioning are adversely affected by iron deficiency in early life. Iron-deficient rats are known to have elevations in extracellular DA and NE, suggesting alterations in reuptake of these monoamines. To explore possible mechanisms by which cellular iron concentrations may alter NE transporter functioning, we utilized NET expressing PC12 cells and iron-deficient rats to explore the relationship between NET protein and mRNA expression patterns and iron concentrations. Treatment of PC12 with the iron chelator, desferrioxamine mesylate (DFO, 50 μM for 24 h), significantly decreased [3H] NE uptake by more than 35% with no apparent change in Km. PC12 cells exposed to increasing concentrations of DFO (25-100 μM) exhibited a dose response decrease in [3H] NE uptake within 24 h (38-73% of control) that paralleled a decrease in cellular NET protein content. Inhibition of protein synthesis with cycloheximide resulted in NET disappearance rates from DFO-treated cells greatly exceeding the rate of loss from control cells. RT-PCR analysis revealed only a modest decrease in NET mRNA levels. Rat brain locus ceruleus and thalamus NET mRNA levels were also only modestly decreased (10-15%) despite a 40% reduction in regional brain iron. In contrast, NET proteins levels in thalamus and locus ceruleus were strongly affected by regional iron deficiency with high correlations with iron concentrations (r > 0.94 and r > 0.80 respectively). The present findings demonstrate that NET protein concentrations and functioning are dramatically reduced with iron deficiency; the modest effect on mRNA levels suggests a stronger influence on NET trafficking and degradation than on protein synthesis.

AB - Neurological development and functioning are adversely affected by iron deficiency in early life. Iron-deficient rats are known to have elevations in extracellular DA and NE, suggesting alterations in reuptake of these monoamines. To explore possible mechanisms by which cellular iron concentrations may alter NE transporter functioning, we utilized NET expressing PC12 cells and iron-deficient rats to explore the relationship between NET protein and mRNA expression patterns and iron concentrations. Treatment of PC12 with the iron chelator, desferrioxamine mesylate (DFO, 50 μM for 24 h), significantly decreased [3H] NE uptake by more than 35% with no apparent change in Km. PC12 cells exposed to increasing concentrations of DFO (25-100 μM) exhibited a dose response decrease in [3H] NE uptake within 24 h (38-73% of control) that paralleled a decrease in cellular NET protein content. Inhibition of protein synthesis with cycloheximide resulted in NET disappearance rates from DFO-treated cells greatly exceeding the rate of loss from control cells. RT-PCR analysis revealed only a modest decrease in NET mRNA levels. Rat brain locus ceruleus and thalamus NET mRNA levels were also only modestly decreased (10-15%) despite a 40% reduction in regional brain iron. In contrast, NET proteins levels in thalamus and locus ceruleus were strongly affected by regional iron deficiency with high correlations with iron concentrations (r > 0.94 and r > 0.80 respectively). The present findings demonstrate that NET protein concentrations and functioning are dramatically reduced with iron deficiency; the modest effect on mRNA levels suggests a stronger influence on NET trafficking and degradation than on protein synthesis.

UR - http://www.scopus.com/inward/record.url?scp=33746884257&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33746884257&partnerID=8YFLogxK

U2 - 10.1016/j.brainres.2006.03.071

DO - 10.1016/j.brainres.2006.03.071

M3 - Article

VL - 1092

SP - 47

EP - 58

JO - Brain Research

JF - Brain Research

SN - 0006-8993

IS - 1

ER -