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Expression and localization of mitochondrial ferritin mRNA in Alzheimer's disease cerebral cortex.

Wang L, Yang H, Zhao S, Sato H, Konishi Y, Beach TG, Abdelalim EM, Bisem NJ, Tooyama I - PLoS ONE (2011)

Bottom Line: The neuroprotective effect of MtF on oxidative stress induced by H(2)O(2) was measured by MTT assay.Real-time PCR and western-blot confirmed that MtF expression levels in the cerebral cortex were significantly higher in AD cases than that in control cases at both the mRNA and the protein level.Finally, MtF expression showed a significant neuroprotective effect against H2O2-induced oxidative stress (p<0.05).

View Article: PubMed Central - PubMed

Affiliation: Molecular Neuroscience Research Center, Shiga University of Medical Science, Otsu, Japan.

ABSTRACT
Mitochondrial ferritin (MtF) has been identified as a novel ferritin encoded by an intron-lacking gene with specific mitochondrial localization located on chromosome 5q23.1. MtF has been associated with neurodegenerative disorders such as Friedreich ataxia and restless leg syndrome. However, little information is available about MtF in Alzheimer's disease (AD). In this study, therefore, we investigated the expression and localization of MtF messenger RNA (mRNA) in the cerebral cortex of AD and control cases using real-time polymerase chain reaction (PCR) as well as in situ hybridization histochemistry. We also examined protein expression using western-blot assay. In addition, we used in vitro methods to further explore the effect of oxidative stress and β-amyloid peptide (Aβ) on MtF expression. To do this we examined MtF mRNA and protein expression changes in the human neuroblastoma cell line, IMR-32, after treatment with Aβ, H2O2, or both. The neuroprotective effect of MtF on oxidative stress induced by H(2)O(2) was measured by MTT assay. The in situ hybridization studies revealed that MtF mRNA was detected mainly in neurons to a lesser degree in glial cells in the cerebral cortex. The staining intensity and the number of positive cells were increased in the cerebral cortex of AD patients. Real-time PCR and western-blot confirmed that MtF expression levels in the cerebral cortex were significantly higher in AD cases than that in control cases at both the mRNA and the protein level. Cell culture experiments demonstrated that the expression of both MtF mRNA and protein were increased by treatment with H2O2 or a combination of Aβ and H2O2, but not with Aβ alone. Finally, MtF expression showed a significant neuroprotective effect against H2O2-induced oxidative stress (p<0.05). The present study suggests that MtF is involved in the pathology of AD and may play a neuroprotective role against oxidative stress.

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Related in: MedlinePlus

Western-blot analysis of MtF expression in each in vitro experiment group.(A) An immunoreactive band of approximately 22 kD probed with an anti-MtF antibody is present in all lanes. (B) When the expression level of MtF is normalized to β-actin, MtF expression after treatment with H2O2 or with a combination of Aβ and H2O2 was significantly greater than that in control cases (**p<0.01. *** p<0.001). However, treatment with Aβ or DMSO alone shows no significant effect on MtF expression.
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pone-0022325-g006: Western-blot analysis of MtF expression in each in vitro experiment group.(A) An immunoreactive band of approximately 22 kD probed with an anti-MtF antibody is present in all lanes. (B) When the expression level of MtF is normalized to β-actin, MtF expression after treatment with H2O2 or with a combination of Aβ and H2O2 was significantly greater than that in control cases (**p<0.01. *** p<0.001). However, treatment with Aβ or DMSO alone shows no significant effect on MtF expression.

Mentions: Figure 6 shows the western-blot analysis of MtF protein expression in the H2O2 treatment group, the Aβ treatment group, the H2O2 plus Aβ treatment group, as well as the two control groups: DMSO treated and untreated cells. MtF protein expression after treatment with H2O2 or a combination of Aβ and H2O2 was significantly higher than in control groups. The combination group showed the highest MtF protein expression level of all groups. However, single treatment with Aβ or DMSO caused no significant effect on MtF protein expression. These results are in accordance with the real-time PCR results.


Expression and localization of mitochondrial ferritin mRNA in Alzheimer's disease cerebral cortex.

Wang L, Yang H, Zhao S, Sato H, Konishi Y, Beach TG, Abdelalim EM, Bisem NJ, Tooyama I - PLoS ONE (2011)

Western-blot analysis of MtF expression in each in vitro experiment group.(A) An immunoreactive band of approximately 22 kD probed with an anti-MtF antibody is present in all lanes. (B) When the expression level of MtF is normalized to β-actin, MtF expression after treatment with H2O2 or with a combination of Aβ and H2O2 was significantly greater than that in control cases (**p<0.01. *** p<0.001). However, treatment with Aβ or DMSO alone shows no significant effect on MtF expression.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3140525&req=5

pone-0022325-g006: Western-blot analysis of MtF expression in each in vitro experiment group.(A) An immunoreactive band of approximately 22 kD probed with an anti-MtF antibody is present in all lanes. (B) When the expression level of MtF is normalized to β-actin, MtF expression after treatment with H2O2 or with a combination of Aβ and H2O2 was significantly greater than that in control cases (**p<0.01. *** p<0.001). However, treatment with Aβ or DMSO alone shows no significant effect on MtF expression.
Mentions: Figure 6 shows the western-blot analysis of MtF protein expression in the H2O2 treatment group, the Aβ treatment group, the H2O2 plus Aβ treatment group, as well as the two control groups: DMSO treated and untreated cells. MtF protein expression after treatment with H2O2 or a combination of Aβ and H2O2 was significantly higher than in control groups. The combination group showed the highest MtF protein expression level of all groups. However, single treatment with Aβ or DMSO caused no significant effect on MtF protein expression. These results are in accordance with the real-time PCR results.

Bottom Line: The neuroprotective effect of MtF on oxidative stress induced by H(2)O(2) was measured by MTT assay.Real-time PCR and western-blot confirmed that MtF expression levels in the cerebral cortex were significantly higher in AD cases than that in control cases at both the mRNA and the protein level.Finally, MtF expression showed a significant neuroprotective effect against H2O2-induced oxidative stress (p<0.05).

View Article: PubMed Central - PubMed

Affiliation: Molecular Neuroscience Research Center, Shiga University of Medical Science, Otsu, Japan.

ABSTRACT
Mitochondrial ferritin (MtF) has been identified as a novel ferritin encoded by an intron-lacking gene with specific mitochondrial localization located on chromosome 5q23.1. MtF has been associated with neurodegenerative disorders such as Friedreich ataxia and restless leg syndrome. However, little information is available about MtF in Alzheimer's disease (AD). In this study, therefore, we investigated the expression and localization of MtF messenger RNA (mRNA) in the cerebral cortex of AD and control cases using real-time polymerase chain reaction (PCR) as well as in situ hybridization histochemistry. We also examined protein expression using western-blot assay. In addition, we used in vitro methods to further explore the effect of oxidative stress and β-amyloid peptide (Aβ) on MtF expression. To do this we examined MtF mRNA and protein expression changes in the human neuroblastoma cell line, IMR-32, after treatment with Aβ, H2O2, or both. The neuroprotective effect of MtF on oxidative stress induced by H(2)O(2) was measured by MTT assay. The in situ hybridization studies revealed that MtF mRNA was detected mainly in neurons to a lesser degree in glial cells in the cerebral cortex. The staining intensity and the number of positive cells were increased in the cerebral cortex of AD patients. Real-time PCR and western-blot confirmed that MtF expression levels in the cerebral cortex were significantly higher in AD cases than that in control cases at both the mRNA and the protein level. Cell culture experiments demonstrated that the expression of both MtF mRNA and protein were increased by treatment with H2O2 or a combination of Aβ and H2O2, but not with Aβ alone. Finally, MtF expression showed a significant neuroprotective effect against H2O2-induced oxidative stress (p<0.05). The present study suggests that MtF is involved in the pathology of AD and may play a neuroprotective role against oxidative stress.

Show MeSH
Related in: MedlinePlus