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The novel protein MANI modulates neurogenesis and neurite-cone growth.

Mishra M, Akatsu H, Heese K - J. Cell. Mol. Med. (2011)

Bottom Line: To date, three myelin-associated proteins [Nogo or reticulon 4 (RTN4), myelin-associated glycoprotein (MAG) and oligodendrocyte myelin glycoprotein (OMG)] are known to inhibit axonal regeneration via activation of the neuronal glycosylphosphatidylinositol-anchored Nogo receptor [NgR, together with p75 neurotrophin receptor (p75NTR) and Lingo-1].We show that knockdown of Cdc27, a component of the anaphase-promoting complex (APC), leads to enhanced neurite outgrowth.Our finding describes the novel MANI-Cdc27-APC pathway as an important cascade that prevents neurons from extending axons, thus providing implications for the potential treatment of neurodegenerative diseases.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cell Biology, School of Biological Sciences, College of Science, Nanyang Technological University, Singapore.

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MANI expression in AD brains. MANI protein expression in the cortex/temporal lobe (the borderline between the cortex and white matter in the temporal lobe) of control subjects (A) and AD patients (B) (representative pictures are shown). IHC was performed with the MANI antibody. No correlation between MANI staining and SPs or NFTs (data not shown) was observed. Scale bar = 200 μm (A, B). Southern blot analysis of MANI RT-PCR expression products in the brains of AD patients and control subjects. Analysis of PCR products; left: Control group, right: AD group; top: MANI PCR products, bottom: PCR products of the reference gene S12. H, hippocampus; Cx, parietal cortex; detailed description of control and AD patients 1–8 is provided in the Supplementary Information, Table S1 (C). Quantitation of the MANI transcripts. Values represent the ratio of the densitometric scores for MANI and S12 PCR products ± S.D. (*P < 0.05 compared with controls) (D). Western blot analysis of Mani protein in an AD mouse model (AD) and control wild-type (litter-mates) (C) total brain lysates confirmed that Mani was significantly decreased in AD. Right: Data represent mean ± S.D. (ratio of the densitometric scores for Mani and Gapdh products) of three independent experiments (*P < 0.05 compared with control) (E). Effect of Mani on App phosphorylation and processing. PC12 cells were grown and differentiated with Ngf, and lysates were prepared on the 12th day of induction, followed by Western blot analysis (c = control (mock-transfected), Mani = Mani transfected). Mani-transfected cells exhibited decreased phosphorylation of processed App (Ctfβ) and increased phosphorylation of full-length App, which in turn stabilizes App’s conformation and inhibits processing (F).
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fig01: MANI expression in AD brains. MANI protein expression in the cortex/temporal lobe (the borderline between the cortex and white matter in the temporal lobe) of control subjects (A) and AD patients (B) (representative pictures are shown). IHC was performed with the MANI antibody. No correlation between MANI staining and SPs or NFTs (data not shown) was observed. Scale bar = 200 μm (A, B). Southern blot analysis of MANI RT-PCR expression products in the brains of AD patients and control subjects. Analysis of PCR products; left: Control group, right: AD group; top: MANI PCR products, bottom: PCR products of the reference gene S12. H, hippocampus; Cx, parietal cortex; detailed description of control and AD patients 1–8 is provided in the Supplementary Information, Table S1 (C). Quantitation of the MANI transcripts. Values represent the ratio of the densitometric scores for MANI and S12 PCR products ± S.D. (*P < 0.05 compared with controls) (D). Western blot analysis of Mani protein in an AD mouse model (AD) and control wild-type (litter-mates) (C) total brain lysates confirmed that Mani was significantly decreased in AD. Right: Data represent mean ± S.D. (ratio of the densitometric scores for Mani and Gapdh products) of three independent experiments (*P < 0.05 compared with control) (E). Effect of Mani on App phosphorylation and processing. PC12 cells were grown and differentiated with Ngf, and lysates were prepared on the 12th day of induction, followed by Western blot analysis (c = control (mock-transfected), Mani = Mani transfected). Mani-transfected cells exhibited decreased phosphorylation of processed App (Ctfβ) and increased phosphorylation of full-length App, which in turn stabilizes App’s conformation and inhibits processing (F).

Mentions: Because we originally found the MANI mRNA to be down-regulated in AD brains, in a cDNA subtraction analysis [4], we reconfirmed this finding by RT-PCR and IHC in human patients and observed a significant reduction of MANI mRNA levels in AD patients. Protein expression analysis in human brains disclosed (though no obvious down-regulation) that MANI was present within (neuronal) axonal fibres and to be associated with myelin sheets (Fig. 1A–D; Fig. S2 [antibody specificity analysis] and Table S1). Because of our limited access to human tissue, we confirmed the down-regulation of Mani at protein levels in AD using an AD mouse model (Fig. 1E).


The novel protein MANI modulates neurogenesis and neurite-cone growth.

Mishra M, Akatsu H, Heese K - J. Cell. Mol. Med. (2011)

MANI expression in AD brains. MANI protein expression in the cortex/temporal lobe (the borderline between the cortex and white matter in the temporal lobe) of control subjects (A) and AD patients (B) (representative pictures are shown). IHC was performed with the MANI antibody. No correlation between MANI staining and SPs or NFTs (data not shown) was observed. Scale bar = 200 μm (A, B). Southern blot analysis of MANI RT-PCR expression products in the brains of AD patients and control subjects. Analysis of PCR products; left: Control group, right: AD group; top: MANI PCR products, bottom: PCR products of the reference gene S12. H, hippocampus; Cx, parietal cortex; detailed description of control and AD patients 1–8 is provided in the Supplementary Information, Table S1 (C). Quantitation of the MANI transcripts. Values represent the ratio of the densitometric scores for MANI and S12 PCR products ± S.D. (*P < 0.05 compared with controls) (D). Western blot analysis of Mani protein in an AD mouse model (AD) and control wild-type (litter-mates) (C) total brain lysates confirmed that Mani was significantly decreased in AD. Right: Data represent mean ± S.D. (ratio of the densitometric scores for Mani and Gapdh products) of three independent experiments (*P < 0.05 compared with control) (E). Effect of Mani on App phosphorylation and processing. PC12 cells were grown and differentiated with Ngf, and lysates were prepared on the 12th day of induction, followed by Western blot analysis (c = control (mock-transfected), Mani = Mani transfected). Mani-transfected cells exhibited decreased phosphorylation of processed App (Ctfβ) and increased phosphorylation of full-length App, which in turn stabilizes App’s conformation and inhibits processing (F).
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fig01: MANI expression in AD brains. MANI protein expression in the cortex/temporal lobe (the borderline between the cortex and white matter in the temporal lobe) of control subjects (A) and AD patients (B) (representative pictures are shown). IHC was performed with the MANI antibody. No correlation between MANI staining and SPs or NFTs (data not shown) was observed. Scale bar = 200 μm (A, B). Southern blot analysis of MANI RT-PCR expression products in the brains of AD patients and control subjects. Analysis of PCR products; left: Control group, right: AD group; top: MANI PCR products, bottom: PCR products of the reference gene S12. H, hippocampus; Cx, parietal cortex; detailed description of control and AD patients 1–8 is provided in the Supplementary Information, Table S1 (C). Quantitation of the MANI transcripts. Values represent the ratio of the densitometric scores for MANI and S12 PCR products ± S.D. (*P < 0.05 compared with controls) (D). Western blot analysis of Mani protein in an AD mouse model (AD) and control wild-type (litter-mates) (C) total brain lysates confirmed that Mani was significantly decreased in AD. Right: Data represent mean ± S.D. (ratio of the densitometric scores for Mani and Gapdh products) of three independent experiments (*P < 0.05 compared with control) (E). Effect of Mani on App phosphorylation and processing. PC12 cells were grown and differentiated with Ngf, and lysates were prepared on the 12th day of induction, followed by Western blot analysis (c = control (mock-transfected), Mani = Mani transfected). Mani-transfected cells exhibited decreased phosphorylation of processed App (Ctfβ) and increased phosphorylation of full-length App, which in turn stabilizes App’s conformation and inhibits processing (F).
Mentions: Because we originally found the MANI mRNA to be down-regulated in AD brains, in a cDNA subtraction analysis [4], we reconfirmed this finding by RT-PCR and IHC in human patients and observed a significant reduction of MANI mRNA levels in AD patients. Protein expression analysis in human brains disclosed (though no obvious down-regulation) that MANI was present within (neuronal) axonal fibres and to be associated with myelin sheets (Fig. 1A–D; Fig. S2 [antibody specificity analysis] and Table S1). Because of our limited access to human tissue, we confirmed the down-regulation of Mani at protein levels in AD using an AD mouse model (Fig. 1E).

Bottom Line: To date, three myelin-associated proteins [Nogo or reticulon 4 (RTN4), myelin-associated glycoprotein (MAG) and oligodendrocyte myelin glycoprotein (OMG)] are known to inhibit axonal regeneration via activation of the neuronal glycosylphosphatidylinositol-anchored Nogo receptor [NgR, together with p75 neurotrophin receptor (p75NTR) and Lingo-1].We show that knockdown of Cdc27, a component of the anaphase-promoting complex (APC), leads to enhanced neurite outgrowth.Our finding describes the novel MANI-Cdc27-APC pathway as an important cascade that prevents neurons from extending axons, thus providing implications for the potential treatment of neurodegenerative diseases.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cell Biology, School of Biological Sciences, College of Science, Nanyang Technological University, Singapore.

Show MeSH
Related in: MedlinePlus