Limits...
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.

Show MeSH

Related in: MedlinePlus

In situ hybridization histochemistry of the cerebral cortex of control (A–C) and Alzheimer's disease (AD) cases (D–F) using antisense (A, B, D, E) and sense (C, F) probes.Mitochondrial ferritin mRNA localizes mainly in neurons. Both the number and intensity of positive neurons increase in AD cases (D) compared to controls (A). Using sense probes (C and F), no signals are detected in the cortex. Bars  = 200 µm in A, C, D, F, and 50 µm in B, E.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3140525&req=5

pone-0022325-g001: In situ hybridization histochemistry of the cerebral cortex of control (A–C) and Alzheimer's disease (AD) cases (D–F) using antisense (A, B, D, E) and sense (C, F) probes.Mitochondrial ferritin mRNA localizes mainly in neurons. Both the number and intensity of positive neurons increase in AD cases (D) compared to controls (A). Using sense probes (C and F), no signals are detected in the cortex. Bars  = 200 µm in A, C, D, F, and 50 µm in B, E.

Mentions: Figure 1 shows the results of in situ hybridization histochemistry in the temporal cortex of a control (Fig. 1A, B, C) and an Alzheimer (Fig. 1D, E, F) case. Positive signals for MtF mRNA were detected in both control (Fig. 1 A) and AD (Fig. 1D) cases using the antisense probe. At a high magnification, positive signals were localized predominantly in neurons (Fig. 1E). The staining intensity in MtF-positive neurons was increased in AD cases (Fig. 1E) compared to controls (Fig. 1B). The sense probe did not generate any signal (Fig. 1C and F).


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)

In situ hybridization histochemistry of the cerebral cortex of control (A–C) and Alzheimer's disease (AD) cases (D–F) using antisense (A, B, D, E) and sense (C, F) probes.Mitochondrial ferritin mRNA localizes mainly in neurons. Both the number and intensity of positive neurons increase in AD cases (D) compared to controls (A). Using sense probes (C and F), no signals are detected in the cortex. Bars  = 200 µm in A, C, D, F, and 50 µm in B, E.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0022325-g001: In situ hybridization histochemistry of the cerebral cortex of control (A–C) and Alzheimer's disease (AD) cases (D–F) using antisense (A, B, D, E) and sense (C, F) probes.Mitochondrial ferritin mRNA localizes mainly in neurons. Both the number and intensity of positive neurons increase in AD cases (D) compared to controls (A). Using sense probes (C and F), no signals are detected in the cortex. Bars  = 200 µm in A, C, D, F, and 50 µm in B, E.
Mentions: Figure 1 shows the results of in situ hybridization histochemistry in the temporal cortex of a control (Fig. 1A, B, C) and an Alzheimer (Fig. 1D, E, F) case. Positive signals for MtF mRNA were detected in both control (Fig. 1 A) and AD (Fig. 1D) cases using the antisense probe. At a high magnification, positive signals were localized predominantly in neurons (Fig. 1E). The staining intensity in MtF-positive neurons was increased in AD cases (Fig. 1E) compared to controls (Fig. 1B). The sense probe did not generate any signal (Fig. 1C and F).

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