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Colocalization of 14-3-3 proteins with SOD1 in Lewy body-like hyaline inclusions in familial amyotrophic lateral sclerosis cases and the animal model.

Okamoto Y, Shirakashi Y, Ihara M, Urushitani M, Oono M, Kawamoto Y, Yamashita H, Shimohama S, Kato S, Hirano A, Tomimoto H, Ito H, Takahashi R - PLoS ONE (2011)

Bottom Line: Cu/Zn superoxide dismutase (SOD1) is a major component of Lewy body-like hyaline inclusion (LBHI) found in the postmortem tissue of SOD1-linked familial amyotrophic lateral sclerosis (FALS) patients.In our recent studies, 14-3-3 proteins have been found in the ubiquitinated inclusions inside the anterior horn cells of spinal cords with sporadic amyotrophic lateral sclerosis (ALS).We examined the postmortem brains and the spinal cords of three FALS cases (A4V SOD1 mutant).

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Kyoto University Graduate School of Medicine, Kyoto, Japan.

ABSTRACT

Background and purpose: Cu/Zn superoxide dismutase (SOD1) is a major component of Lewy body-like hyaline inclusion (LBHI) found in the postmortem tissue of SOD1-linked familial amyotrophic lateral sclerosis (FALS) patients. In our recent studies, 14-3-3 proteins have been found in the ubiquitinated inclusions inside the anterior horn cells of spinal cords with sporadic amyotrophic lateral sclerosis (ALS). To further investigate the role of 14-3-3 proteins in ALS, we performed immunohistochemical analysis of 14-3-3 proteins and compared their distributions with those of SOD1 in FALS patients and SOD1-overexpressing mice.

Methods: We examined the postmortem brains and the spinal cords of three FALS cases (A4V SOD1 mutant). Transgenic mice expressing the G93A mutant human SOD1 (mutant SOD1-Tg mice), transgenic mice expressing the wild-type human SOD1 (wild-type SOD1-Tg mice), and non-Tg wild-type mice were also subjected to the immunohistochemical analysis.

Results: In all the FALS patients, LBHIs were observed in the cytoplasm of the anterior horn cells, and these inclusions were immunopositive intensely for pan 14-3-3, 14-3-3β, and 14-3-3γ. In the mutant SOD1-Tg mice, a high degree of immunoreactivity for misfolded SOD1 (C4F6) was observed in the cytoplasm, with an even greater degree of immunoreactivity present in the cytoplasmic aggregates of the anterior horn cells in the lumbar spinal cord. Furthermore, we have found increased 14-3-3β and 14-3-3γ immunoreactivities in the mutant SOD1-Tg mice. Double immunofluorescent staining showed that C4F6 and 14-3-3 proteins were partially co-localized in the spinal cord with FALS and the mutant SOD1-Tg mice. In comparison, the wild-type SOD1-Tg and non-Tg wild-type mice showed no or faint immunoreactivity for C4F6 and 14-3-3 proteins (pan 14-3-3, 14-3-3β, and 14-3-3γ) in any neuronal compartments.

Discussion: These results suggest that 14-3-3 proteins may be associated with the formation of SOD1-containing inclusions, in FALS patients and the mutant SOD1-Tg mice.

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Related in: MedlinePlus

LBHIs immunopositive for 14-3-3 proteins in FALS patients.A, B, and C are the same sections as D, E, and F, respectively. The upper panels (A–C) are stained with H&E, and the lower panels (D–F) are immunostained with the anti-pan 14-3-3 antibody. LBHIs observed on H&E in the anterior horn cells are intensely immunopositive for pan 14-3-3. Bar indicates 100 µm in (A, D), and 50 µm in (B, C, E, F).
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pone-0020427-g001: LBHIs immunopositive for 14-3-3 proteins in FALS patients.A, B, and C are the same sections as D, E, and F, respectively. The upper panels (A–C) are stained with H&E, and the lower panels (D–F) are immunostained with the anti-pan 14-3-3 antibody. LBHIs observed on H&E in the anterior horn cells are intensely immunopositive for pan 14-3-3. Bar indicates 100 µm in (A, D), and 50 µm in (B, C, E, F).

Mentions: In all the three FALS cases, LBHIs were observed inside the anterior horn cells (Figure 1A–C). All the LBHIs observed on H&E showed strong pan 14-3-3 immunoreactivity (Figure 1D–F). Using 14-3-3 isoform-specific antibodies, all the LBHIs detected by H&E (Figure 2A, B) were intensely immunopositive both for 14-3-3β (Figure 2C) and 14-3-3γ (Figure 2D).


Colocalization of 14-3-3 proteins with SOD1 in Lewy body-like hyaline inclusions in familial amyotrophic lateral sclerosis cases and the animal model.

Okamoto Y, Shirakashi Y, Ihara M, Urushitani M, Oono M, Kawamoto Y, Yamashita H, Shimohama S, Kato S, Hirano A, Tomimoto H, Ito H, Takahashi R - PLoS ONE (2011)

LBHIs immunopositive for 14-3-3 proteins in FALS patients.A, B, and C are the same sections as D, E, and F, respectively. The upper panels (A–C) are stained with H&E, and the lower panels (D–F) are immunostained with the anti-pan 14-3-3 antibody. LBHIs observed on H&E in the anterior horn cells are intensely immunopositive for pan 14-3-3. Bar indicates 100 µm in (A, D), and 50 µm in (B, C, E, F).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3105059&req=5

pone-0020427-g001: LBHIs immunopositive for 14-3-3 proteins in FALS patients.A, B, and C are the same sections as D, E, and F, respectively. The upper panels (A–C) are stained with H&E, and the lower panels (D–F) are immunostained with the anti-pan 14-3-3 antibody. LBHIs observed on H&E in the anterior horn cells are intensely immunopositive for pan 14-3-3. Bar indicates 100 µm in (A, D), and 50 µm in (B, C, E, F).
Mentions: In all the three FALS cases, LBHIs were observed inside the anterior horn cells (Figure 1A–C). All the LBHIs observed on H&E showed strong pan 14-3-3 immunoreactivity (Figure 1D–F). Using 14-3-3 isoform-specific antibodies, all the LBHIs detected by H&E (Figure 2A, B) were intensely immunopositive both for 14-3-3β (Figure 2C) and 14-3-3γ (Figure 2D).

Bottom Line: Cu/Zn superoxide dismutase (SOD1) is a major component of Lewy body-like hyaline inclusion (LBHI) found in the postmortem tissue of SOD1-linked familial amyotrophic lateral sclerosis (FALS) patients.In our recent studies, 14-3-3 proteins have been found in the ubiquitinated inclusions inside the anterior horn cells of spinal cords with sporadic amyotrophic lateral sclerosis (ALS).We examined the postmortem brains and the spinal cords of three FALS cases (A4V SOD1 mutant).

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Kyoto University Graduate School of Medicine, Kyoto, Japan.

ABSTRACT

Background and purpose: Cu/Zn superoxide dismutase (SOD1) is a major component of Lewy body-like hyaline inclusion (LBHI) found in the postmortem tissue of SOD1-linked familial amyotrophic lateral sclerosis (FALS) patients. In our recent studies, 14-3-3 proteins have been found in the ubiquitinated inclusions inside the anterior horn cells of spinal cords with sporadic amyotrophic lateral sclerosis (ALS). To further investigate the role of 14-3-3 proteins in ALS, we performed immunohistochemical analysis of 14-3-3 proteins and compared their distributions with those of SOD1 in FALS patients and SOD1-overexpressing mice.

Methods: We examined the postmortem brains and the spinal cords of three FALS cases (A4V SOD1 mutant). Transgenic mice expressing the G93A mutant human SOD1 (mutant SOD1-Tg mice), transgenic mice expressing the wild-type human SOD1 (wild-type SOD1-Tg mice), and non-Tg wild-type mice were also subjected to the immunohistochemical analysis.

Results: In all the FALS patients, LBHIs were observed in the cytoplasm of the anterior horn cells, and these inclusions were immunopositive intensely for pan 14-3-3, 14-3-3β, and 14-3-3γ. In the mutant SOD1-Tg mice, a high degree of immunoreactivity for misfolded SOD1 (C4F6) was observed in the cytoplasm, with an even greater degree of immunoreactivity present in the cytoplasmic aggregates of the anterior horn cells in the lumbar spinal cord. Furthermore, we have found increased 14-3-3β and 14-3-3γ immunoreactivities in the mutant SOD1-Tg mice. Double immunofluorescent staining showed that C4F6 and 14-3-3 proteins were partially co-localized in the spinal cord with FALS and the mutant SOD1-Tg mice. In comparison, the wild-type SOD1-Tg and non-Tg wild-type mice showed no or faint immunoreactivity for C4F6 and 14-3-3 proteins (pan 14-3-3, 14-3-3β, and 14-3-3γ) in any neuronal compartments.

Discussion: These results suggest that 14-3-3 proteins may be associated with the formation of SOD1-containing inclusions, in FALS patients and the mutant SOD1-Tg mice.

Show MeSH
Related in: MedlinePlus