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Disease-Modifying Effect of Adiponectin in Model of α-Synucleinopathies.

Sekiyama K, Waragai M, Akatsu H, Sugama S, Takenouchi T, Takamatsu Y, Fujita M, Sekigawa A, Rockenstein E, Inoue S, La Spada AR, Masliah E, Hashimoto M - Ann Clin Transl Neurol (2014)

Bottom Line: In neuronal cells expressing α-synuclein (αS), aggregation of αS was suppressed by treatment with recombinant APN in an AdipoRI-AMP kinase pathway-dependent manner.Concomitantly, phosphorylation and release of αS were significantly decreased by APN, suggesting that APN may be antineurodegenerative.Taken together, APN may suppress neurodegeneration through modification of the metabolic pathway, and could possess a therapeutic potential against α-synucleinopathies.

View Article: PubMed Central - PubMed

Affiliation: Tokyo Metropolitan Institute of Medical Sciences, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo 156-0057, Japan.

ABSTRACT

Objective: Growing evidence suggests that neurodegenerative diseases are associated with metabolic disorders, but the mechanisms are still unclear. Better comprehension of this issue might provide a new strategy for treatment of neurodegenerative diseases. We investigated possible roles of adiponectin (APN), the anti-diabetes protein, in the pathogenesis of α-synucleinopathies.

Methods: Using biochemical and histological methods, we investigated autopsy brain of α-synucleinopathies including Parkinson's disease (PD) and dementia with Lewy bodies (DLB), and analyzed the effects of APN in cellular and in mouse models of α-synucleinopathies.

Results: We observed that APN is localized in Lewy bodies derived from α-synucleinopathies such as Parkinson's disease and dementia with Lewy bodies. In neuronal cells expressing α-synuclein (αS), aggregation of αS was suppressed by treatment with recombinant APN in an AdipoRI-AMP kinase pathway-dependent manner. Concomitantly, phosphorylation and release of αS were significantly decreased by APN, suggesting that APN may be antineurodegenerative. In transgenic mice expressing αS, both histopathology and movement disorder were significantly improved by intranasal treatment with globular APN when the treatment was initiated in the early stage of the disease. In a mouse model, reduced levels of guanosine- and inosine- monophosphates, both of which are potential stimulators of aggregation of αS, might partly contribute to suppression of aggregation of αS by APN.

Interpretation: Taken together, APN may suppress neurodegeneration through modification of the metabolic pathway, and could possess a therapeutic potential against α-synucleinopathies.

No MeSH data available.


Related in: MedlinePlus

APN ameliorates neurodegeneration in a mouse model of α-synucleinopathies. gAPN (0.1 mg/mL in 10 μL PBS) or PBS alone (10 μL) was injected into the nasal cavities of αS tg mice (male, 3-month-old) or wild-type littermates every 3 days for 2 months. Body weight was measured (A) and motor performances were evaluated by rotarod test (B) and beam test (C) (mean ± SEM, n = 8–9, *P < 0.05, **P < 0.01, ***P < 0.001). Mice brains were then analyzed histologically and biochemically. (D) Representative immunohistochemical images: the cortex and olfactory bulb were stained with anti-pαS, and the thalamus was probed with anti-αS. Insets are shown to show at higher magnification for the cortex. (E) Representative images of immunoblotting (cortex). The intensities of αS in the FA fraction were quantified (mean ± SEM, n = 6, **P < 0.01). FA, formic acid; APN, adiponectin.
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fig03: APN ameliorates neurodegeneration in a mouse model of α-synucleinopathies. gAPN (0.1 mg/mL in 10 μL PBS) or PBS alone (10 μL) was injected into the nasal cavities of αS tg mice (male, 3-month-old) or wild-type littermates every 3 days for 2 months. Body weight was measured (A) and motor performances were evaluated by rotarod test (B) and beam test (C) (mean ± SEM, n = 8–9, *P < 0.05, **P < 0.01, ***P < 0.001). Mice brains were then analyzed histologically and biochemically. (D) Representative immunohistochemical images: the cortex and olfactory bulb were stained with anti-pαS, and the thalamus was probed with anti-αS. Insets are shown to show at higher magnification for the cortex. (E) Representative images of immunoblotting (cortex). The intensities of αS in the FA fraction were quantified (mean ± SEM, n = 6, **P < 0.01). FA, formic acid; APN, adiponectin.

Mentions: On the basis of the findings in the autopsy brains and cell-based study, we wished to investigate the effect of APN on α-synucleinopathies in vivo. For this purpose, recombinant globular domain of APN (gAPN), a physiological product of C-terminal APN,11 was administered intranasally to tg mice expressing αS (Fig.3). We used gAPN not simply because gAPN is shorter than full length of APN but because suppressive effect of gAPN on the aggregation of αS in cell cultures was comparable to that of full length of APN (Fig. S3A). Furthermore, intranasally injected FLAG-tagged gAPN was shown to reach various areas of the brain in mice (Fig. S3B and C). Indeed, such a noninvasive delivery of therapeutics directly to the brain has been considered for many reagents, including insulin treatment for dementia.31 With this background, the APN nasal treatment was initiated in the αS tg mice at 3 months of age without severe movement disorder (Fig.3, male, n = 8–9). Compared to the mice that did not receive gAPN, the mice at 5 months of age that received gAPN had significantly improved weight loss (Fig.3A) and retarded the progression of movement disability, as assessed by a rotarod test (Fig.3B), followed by an examination by beam walking assay (Fig.3C). In addition to the behavior analyses, αS pathology was remarkably ameliorated in the mice treated with gAPN compared to the mice that received vehicle injection as evaluated by immunohistochemistry; accumulation of pαS was reduced in both cortex and olfactory bulb, while αS-positive globule formation in thalamus was remarkably suppressed (Fig.3D). Furthermore, these results were confirmed by immunoblot analysis showing that accumulation of αS in the FA-extractable fractions of the cortex of the αS tg mice brain was remarkably suppressed by treatment with gAPN (Fig.3E). During the experimental periods, the activation of glial cells, including astrocytes and microglia, was not extensive as to be quantified (not shown). In contrast to the current protocol of experiment, movement disorder was little improved when the intranasal treatment of APN was initiated in the αS tg mice at 5 months of age which had severer movement disorders compared to the mice at 3 months of age (not shown). Taken together, these results suggest that intranasal gAPN protects against neurodegeneration in the early stage in the αS tg mouse.


Disease-Modifying Effect of Adiponectin in Model of α-Synucleinopathies.

Sekiyama K, Waragai M, Akatsu H, Sugama S, Takenouchi T, Takamatsu Y, Fujita M, Sekigawa A, Rockenstein E, Inoue S, La Spada AR, Masliah E, Hashimoto M - Ann Clin Transl Neurol (2014)

APN ameliorates neurodegeneration in a mouse model of α-synucleinopathies. gAPN (0.1 mg/mL in 10 μL PBS) or PBS alone (10 μL) was injected into the nasal cavities of αS tg mice (male, 3-month-old) or wild-type littermates every 3 days for 2 months. Body weight was measured (A) and motor performances were evaluated by rotarod test (B) and beam test (C) (mean ± SEM, n = 8–9, *P < 0.05, **P < 0.01, ***P < 0.001). Mice brains were then analyzed histologically and biochemically. (D) Representative immunohistochemical images: the cortex and olfactory bulb were stained with anti-pαS, and the thalamus was probed with anti-αS. Insets are shown to show at higher magnification for the cortex. (E) Representative images of immunoblotting (cortex). The intensities of αS in the FA fraction were quantified (mean ± SEM, n = 6, **P < 0.01). FA, formic acid; APN, adiponectin.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4128281&req=5

fig03: APN ameliorates neurodegeneration in a mouse model of α-synucleinopathies. gAPN (0.1 mg/mL in 10 μL PBS) or PBS alone (10 μL) was injected into the nasal cavities of αS tg mice (male, 3-month-old) or wild-type littermates every 3 days for 2 months. Body weight was measured (A) and motor performances were evaluated by rotarod test (B) and beam test (C) (mean ± SEM, n = 8–9, *P < 0.05, **P < 0.01, ***P < 0.001). Mice brains were then analyzed histologically and biochemically. (D) Representative immunohistochemical images: the cortex and olfactory bulb were stained with anti-pαS, and the thalamus was probed with anti-αS. Insets are shown to show at higher magnification for the cortex. (E) Representative images of immunoblotting (cortex). The intensities of αS in the FA fraction were quantified (mean ± SEM, n = 6, **P < 0.01). FA, formic acid; APN, adiponectin.
Mentions: On the basis of the findings in the autopsy brains and cell-based study, we wished to investigate the effect of APN on α-synucleinopathies in vivo. For this purpose, recombinant globular domain of APN (gAPN), a physiological product of C-terminal APN,11 was administered intranasally to tg mice expressing αS (Fig.3). We used gAPN not simply because gAPN is shorter than full length of APN but because suppressive effect of gAPN on the aggregation of αS in cell cultures was comparable to that of full length of APN (Fig. S3A). Furthermore, intranasally injected FLAG-tagged gAPN was shown to reach various areas of the brain in mice (Fig. S3B and C). Indeed, such a noninvasive delivery of therapeutics directly to the brain has been considered for many reagents, including insulin treatment for dementia.31 With this background, the APN nasal treatment was initiated in the αS tg mice at 3 months of age without severe movement disorder (Fig.3, male, n = 8–9). Compared to the mice that did not receive gAPN, the mice at 5 months of age that received gAPN had significantly improved weight loss (Fig.3A) and retarded the progression of movement disability, as assessed by a rotarod test (Fig.3B), followed by an examination by beam walking assay (Fig.3C). In addition to the behavior analyses, αS pathology was remarkably ameliorated in the mice treated with gAPN compared to the mice that received vehicle injection as evaluated by immunohistochemistry; accumulation of pαS was reduced in both cortex and olfactory bulb, while αS-positive globule formation in thalamus was remarkably suppressed (Fig.3D). Furthermore, these results were confirmed by immunoblot analysis showing that accumulation of αS in the FA-extractable fractions of the cortex of the αS tg mice brain was remarkably suppressed by treatment with gAPN (Fig.3E). During the experimental periods, the activation of glial cells, including astrocytes and microglia, was not extensive as to be quantified (not shown). In contrast to the current protocol of experiment, movement disorder was little improved when the intranasal treatment of APN was initiated in the αS tg mice at 5 months of age which had severer movement disorders compared to the mice at 3 months of age (not shown). Taken together, these results suggest that intranasal gAPN protects against neurodegeneration in the early stage in the αS tg mouse.

Bottom Line: In neuronal cells expressing α-synuclein (αS), aggregation of αS was suppressed by treatment with recombinant APN in an AdipoRI-AMP kinase pathway-dependent manner.Concomitantly, phosphorylation and release of αS were significantly decreased by APN, suggesting that APN may be antineurodegenerative.Taken together, APN may suppress neurodegeneration through modification of the metabolic pathway, and could possess a therapeutic potential against α-synucleinopathies.

View Article: PubMed Central - PubMed

Affiliation: Tokyo Metropolitan Institute of Medical Sciences, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo 156-0057, Japan.

ABSTRACT

Objective: Growing evidence suggests that neurodegenerative diseases are associated with metabolic disorders, but the mechanisms are still unclear. Better comprehension of this issue might provide a new strategy for treatment of neurodegenerative diseases. We investigated possible roles of adiponectin (APN), the anti-diabetes protein, in the pathogenesis of α-synucleinopathies.

Methods: Using biochemical and histological methods, we investigated autopsy brain of α-synucleinopathies including Parkinson's disease (PD) and dementia with Lewy bodies (DLB), and analyzed the effects of APN in cellular and in mouse models of α-synucleinopathies.

Results: We observed that APN is localized in Lewy bodies derived from α-synucleinopathies such as Parkinson's disease and dementia with Lewy bodies. In neuronal cells expressing α-synuclein (αS), aggregation of αS was suppressed by treatment with recombinant APN in an AdipoRI-AMP kinase pathway-dependent manner. Concomitantly, phosphorylation and release of αS were significantly decreased by APN, suggesting that APN may be antineurodegenerative. In transgenic mice expressing αS, both histopathology and movement disorder were significantly improved by intranasal treatment with globular APN when the treatment was initiated in the early stage of the disease. In a mouse model, reduced levels of guanosine- and inosine- monophosphates, both of which are potential stimulators of aggregation of αS, might partly contribute to suppression of aggregation of αS by APN.

Interpretation: Taken together, APN may suppress neurodegeneration through modification of the metabolic pathway, and could possess a therapeutic potential against α-synucleinopathies.

No MeSH data available.


Related in: MedlinePlus