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Up-regulation of neural and cell cycle-related microRNAs in brain of amyotrophic lateral sclerosis mice at late disease stage.

Marcuzzo S, Bonanno S, Kapetis D, Barzago C, Cavalcante P, D'Alessandro S, Mantegazza R, Bernasconi P - Mol Brain (2015)

Bottom Line: Expression of miR-9, miR-124a, miR-19a and -19b was significantly increased in G93A-SOD1 whole brain at late stage disease compared to B6.SJL and Wt-SOD1 control brains.In G93A-SOD1 brainstem motor nuclei and primary motor cortex, miR-9 and miR-124a were significantly up-regulated, miR-125b expression was also increased. miR-19a and -19b were up-regulated in primary motor cortex and hippocampus, respectively.Expression analysis of predicted miRNA targets identified miRNA/target gene pairs differentially expressed in G93A-SOD1 brain regions compared to controls.

View Article: PubMed Central - PubMed

Affiliation: Neurology IV - Neuromuscular Diseases and Neuroimmunology Unit, Fondazione Istituto Neurologico "Carlo Besta", Via Celoria 11, Milan, 20133, Italy. stefania.marcuzzo@istituto-besta.it.

ABSTRACT

Background: Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by selective motor neuron degeneration in motor cortex, brainstem and spinal cord. microRNAs (miRNAs) are small non-coding RNAs that bind complementary target sequences and modulate gene expression; they are key molecules for establishing a neuronal phenotype, and in neurodegeneration. Here we investigated neural miR-9, miR-124a, miR-125b, miR-219, miR-134, and cell cycle-related miR-19a and -19b, in G93A-SOD1 mouse brain in pre-symptomatic and late stage disease.

Results: Expression of miR-9, miR-124a, miR-19a and -19b was significantly increased in G93A-SOD1 whole brain at late stage disease compared to B6.SJL and Wt-SOD1 control brains. These miRNAs were then analyzed in manually dissected SVZ, hippocampus, primary motor cortex and brainstem motor nuclei in 18-week-old ALS mice compared to same age controls. In SVZ and hippocampus miR-124a was up-regulated, miR-219 was down-regulated, and numbers of neural stem progenitor cells (NSPCs) were significantly increased. In G93A-SOD1 brainstem motor nuclei and primary motor cortex, miR-9 and miR-124a were significantly up-regulated, miR-125b expression was also increased. miR-19a and -19b were up-regulated in primary motor cortex and hippocampus, respectively. Expression analysis of predicted miRNA targets identified miRNA/target gene pairs differentially expressed in G93A-SOD1 brain regions compared to controls.

Conclusions: Hierarchical clustering analysis, identifying two clusters of miRNA/target genes, one characterizing brainstem motor nuclei and primary motor cortex, the other hippocampus and SVZ, suggests that altered expression of neural and cell cycle-related miRNAs in these brain regions might contribute to ALS pathogenesis in G93A-SOD1 mice. Re-establishing their expression to normal levels could be a new therapeutic approach to ALS.

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Brain-specific, cell cycle- and glia-related miRNA expression is altered compared to control in different regions of G93A-SOD1 brain. RT-PCR analysis of brain-specific (A), cell cycle- (B) and glia- (C) related miRNAs in subventricular zone (SVZ), hippocampus (H), primary motor cortex (MC), and brainstem motor nuclei (MN) of G93A-SOD1 and Wt-SOD1 brain at late stage disease. Data are means of relative expression ± SD of 5 mice per group. *p < 0.05; **p < 0.01; limma moderated t-test.
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Fig2: Brain-specific, cell cycle- and glia-related miRNA expression is altered compared to control in different regions of G93A-SOD1 brain. RT-PCR analysis of brain-specific (A), cell cycle- (B) and glia- (C) related miRNAs in subventricular zone (SVZ), hippocampus (H), primary motor cortex (MC), and brainstem motor nuclei (MN) of G93A-SOD1 and Wt-SOD1 brain at late stage disease. Data are means of relative expression ± SD of 5 mice per group. *p < 0.05; **p < 0.01; limma moderated t-test.

Mentions: We next wondered whether altered miRNA expression in the brain of 18-week-old ALS mice would be evident in specific brain regions concerned with neurogenesis (SVZ and hippocampus) and affected by ALS (primary motor cortex and brainstem motor nuclei). We also analyzed miR-125b, marker of reactive astrocytes [29], and miR-219, marker of oligodendrocyte differentiation [30], to investigate glial cell involvement in these brain regions. We found that miR-124a expression was significantly greater in SVZ, hippocampus, primary motor cortex, and brainstem motor nuclei of ALS than Wt-SOD1 mice (p < 0.01). miR-9 expression was lower in SVZ and hippocampus, and significantly greater in primary motor cortex and brainstem motor nuclei (p < 0.01) in ALS compared to control (Figure 2A). miR-19a was significantly down-regulated in hippocampus and brainstem motor nuclei and significantly up-regulated in primary motor cortex of G93A-SOD1 compared to control (p < 0.01). miR-19b was up-regulated in hippocampus (p < 0.05) and down-regulated in primary motor cortex and brainstem motor nuclei (p < 0.05 and p < 0.01, respectively) (Figure 2B). miR-125b was significantly down-regulated in G93A-SOD1 SVZ and hippocampus (p < 0.01) and significantly up-regulated in primary motor cortex (p < 0.01). Finally, miR-219 was significantly lower in ALS SVZ than control SVZ (p < 0.01) (Figure 2C). miR-125b and miR-219 tended to be expressed at higher levels (not significant) in cervical, thoracic and lumbar spinal cord of ALS than control mice (Additional file 2: Figure S2).Figure 2


Up-regulation of neural and cell cycle-related microRNAs in brain of amyotrophic lateral sclerosis mice at late disease stage.

Marcuzzo S, Bonanno S, Kapetis D, Barzago C, Cavalcante P, D'Alessandro S, Mantegazza R, Bernasconi P - Mol Brain (2015)

Brain-specific, cell cycle- and glia-related miRNA expression is altered compared to control in different regions of G93A-SOD1 brain. RT-PCR analysis of brain-specific (A), cell cycle- (B) and glia- (C) related miRNAs in subventricular zone (SVZ), hippocampus (H), primary motor cortex (MC), and brainstem motor nuclei (MN) of G93A-SOD1 and Wt-SOD1 brain at late stage disease. Data are means of relative expression ± SD of 5 mice per group. *p < 0.05; **p < 0.01; limma moderated t-test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4318136&req=5

Fig2: Brain-specific, cell cycle- and glia-related miRNA expression is altered compared to control in different regions of G93A-SOD1 brain. RT-PCR analysis of brain-specific (A), cell cycle- (B) and glia- (C) related miRNAs in subventricular zone (SVZ), hippocampus (H), primary motor cortex (MC), and brainstem motor nuclei (MN) of G93A-SOD1 and Wt-SOD1 brain at late stage disease. Data are means of relative expression ± SD of 5 mice per group. *p < 0.05; **p < 0.01; limma moderated t-test.
Mentions: We next wondered whether altered miRNA expression in the brain of 18-week-old ALS mice would be evident in specific brain regions concerned with neurogenesis (SVZ and hippocampus) and affected by ALS (primary motor cortex and brainstem motor nuclei). We also analyzed miR-125b, marker of reactive astrocytes [29], and miR-219, marker of oligodendrocyte differentiation [30], to investigate glial cell involvement in these brain regions. We found that miR-124a expression was significantly greater in SVZ, hippocampus, primary motor cortex, and brainstem motor nuclei of ALS than Wt-SOD1 mice (p < 0.01). miR-9 expression was lower in SVZ and hippocampus, and significantly greater in primary motor cortex and brainstem motor nuclei (p < 0.01) in ALS compared to control (Figure 2A). miR-19a was significantly down-regulated in hippocampus and brainstem motor nuclei and significantly up-regulated in primary motor cortex of G93A-SOD1 compared to control (p < 0.01). miR-19b was up-regulated in hippocampus (p < 0.05) and down-regulated in primary motor cortex and brainstem motor nuclei (p < 0.05 and p < 0.01, respectively) (Figure 2B). miR-125b was significantly down-regulated in G93A-SOD1 SVZ and hippocampus (p < 0.01) and significantly up-regulated in primary motor cortex (p < 0.01). Finally, miR-219 was significantly lower in ALS SVZ than control SVZ (p < 0.01) (Figure 2C). miR-125b and miR-219 tended to be expressed at higher levels (not significant) in cervical, thoracic and lumbar spinal cord of ALS than control mice (Additional file 2: Figure S2).Figure 2

Bottom Line: Expression of miR-9, miR-124a, miR-19a and -19b was significantly increased in G93A-SOD1 whole brain at late stage disease compared to B6.SJL and Wt-SOD1 control brains.In G93A-SOD1 brainstem motor nuclei and primary motor cortex, miR-9 and miR-124a were significantly up-regulated, miR-125b expression was also increased. miR-19a and -19b were up-regulated in primary motor cortex and hippocampus, respectively.Expression analysis of predicted miRNA targets identified miRNA/target gene pairs differentially expressed in G93A-SOD1 brain regions compared to controls.

View Article: PubMed Central - PubMed

Affiliation: Neurology IV - Neuromuscular Diseases and Neuroimmunology Unit, Fondazione Istituto Neurologico "Carlo Besta", Via Celoria 11, Milan, 20133, Italy. stefania.marcuzzo@istituto-besta.it.

ABSTRACT

Background: Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by selective motor neuron degeneration in motor cortex, brainstem and spinal cord. microRNAs (miRNAs) are small non-coding RNAs that bind complementary target sequences and modulate gene expression; they are key molecules for establishing a neuronal phenotype, and in neurodegeneration. Here we investigated neural miR-9, miR-124a, miR-125b, miR-219, miR-134, and cell cycle-related miR-19a and -19b, in G93A-SOD1 mouse brain in pre-symptomatic and late stage disease.

Results: Expression of miR-9, miR-124a, miR-19a and -19b was significantly increased in G93A-SOD1 whole brain at late stage disease compared to B6.SJL and Wt-SOD1 control brains. These miRNAs were then analyzed in manually dissected SVZ, hippocampus, primary motor cortex and brainstem motor nuclei in 18-week-old ALS mice compared to same age controls. In SVZ and hippocampus miR-124a was up-regulated, miR-219 was down-regulated, and numbers of neural stem progenitor cells (NSPCs) were significantly increased. In G93A-SOD1 brainstem motor nuclei and primary motor cortex, miR-9 and miR-124a were significantly up-regulated, miR-125b expression was also increased. miR-19a and -19b were up-regulated in primary motor cortex and hippocampus, respectively. Expression analysis of predicted miRNA targets identified miRNA/target gene pairs differentially expressed in G93A-SOD1 brain regions compared to controls.

Conclusions: Hierarchical clustering analysis, identifying two clusters of miRNA/target genes, one characterizing brainstem motor nuclei and primary motor cortex, the other hippocampus and SVZ, suggests that altered expression of neural and cell cycle-related miRNAs in these brain regions might contribute to ALS pathogenesis in G93A-SOD1 mice. Re-establishing their expression to normal levels could be a new therapeutic approach to ALS.

Show MeSH
Related in: MedlinePlus