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Role of glial 14-3-3 gamma protein in autoimmune demyelination.

Lee DH, Steinacker P, Seubert S, Turnescu T, Melms A, Manzel A, Otto M, Linker RA - J Neuroinflammation (2015)

Bottom Line: At the same time, deficiency in 14-3-3 γ protein did not influence the immune response.Further histological studies revealed an increased susceptibility towards apoptosis in 14-3-3 γ-deficient OL in the inflamed spinal cord.These data argue for a pivotal role of 14-3-3 γ-mediated signalling pathways for OL protection in neuroinflammation.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Friedrich-Alexander-University Erlangen-Nuremberg, Schwabachanlage 6, D-91054, Erlangen, Germany.

ABSTRACT

Background: The family of 14-3-3 proteins plays an important role in the regulation of cell survival and death. Here, we investigate the role of the 14-3-3 gamma (14-3-3 γ) subunit for glial responses in autoimmune demyelination.

Methods: Expression of 14-3-3 γ in glial cell culture was investigated by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry. 14-3-3 γ knockout mice were subjected to murine myelin oligodendrocyte-induced experimental autoimmune encephalomyelitis (MOG-EAE), an animal model mimicking inflammatory features and neurodegenerative aspects of multiple sclerosis (MS).

Results: Expression studies in cell culture confined expression of 14-3-3 γ to both, oligodendrocytes (OL) and astrocytes. RT-PCR analysis revealed an increased expression of 14-3-3 γ mRNA in the spinal cord during the late chronic phase of MOG-EAE. At that stage, EAE was more severe in 14-3-3 γ knockout mice as compared to age- and gender-matched controls. Histopathological analyses on day 56 post immunization (p.i.) revealed significantly enhanced myelin damage as well as OL injury and secondary, an increase in axonal injury and gliosis in 14-3-3 γ -/- mice. At the same time, deficiency in 14-3-3 γ protein did not influence the immune response. Further histological studies revealed an increased susceptibility towards apoptosis in 14-3-3 γ-deficient OL in the inflamed spinal cord.

Conclusion: These data argue for a pivotal role of 14-3-3 γ-mediated signalling pathways for OL protection in neuroinflammation.

No MeSH data available.


Related in: MedlinePlus

Expression of 14-3-3 γ in glial cells. a RT-PCR analyses of spinal cord for the presence of 14-3-3 γ mRNA (n = 4 mice per time point). The relative expression in the spinal cord of naïve mice is set to 1. Note the increase of 14-3-3 γ expression in the spinal cord during EAE, particularly at later time points (*p < 0.05 at day 78 of EAE). b RT-PCR analysis of 14-3-3 γ mRNA in glial cells as compared to naïve spinal cord (n = 4 for spinal cord versus n = 9 independent cell preparations for astrocytes and n = 3 independent preparations for OPC). Note the significant 14-3-3 γ mRNA expression in glial cells as compared to total spinal cord (**p < 0.01). c, d Representative confocal laser scanning images after immunolabeling for 14-3-3 γ with GFAP in primary astrocyte culture c or Olig2 in primary OPC culture d shows expression of 14-3-3 γ on the protein level in the cytoplasm of some, but not all cells (arrows depict double labelled cells, bar = 20 μm)
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Fig1: Expression of 14-3-3 γ in glial cells. a RT-PCR analyses of spinal cord for the presence of 14-3-3 γ mRNA (n = 4 mice per time point). The relative expression in the spinal cord of naïve mice is set to 1. Note the increase of 14-3-3 γ expression in the spinal cord during EAE, particularly at later time points (*p < 0.05 at day 78 of EAE). b RT-PCR analysis of 14-3-3 γ mRNA in glial cells as compared to naïve spinal cord (n = 4 for spinal cord versus n = 9 independent cell preparations for astrocytes and n = 3 independent preparations for OPC). Note the significant 14-3-3 γ mRNA expression in glial cells as compared to total spinal cord (**p < 0.01). c, d Representative confocal laser scanning images after immunolabeling for 14-3-3 γ with GFAP in primary astrocyte culture c or Olig2 in primary OPC culture d shows expression of 14-3-3 γ on the protein level in the cytoplasm of some, but not all cells (arrows depict double labelled cells, bar = 20 μm)

Mentions: First, we were interested whether 14-3-3 γ is expressed in EAE. To this end, we compared 14-3-3 γ expression in naïve spinal cord to inflamed spinal cord at different time points of EAE (maximum of disease, day 18 or chronic phase of EAE, day 50 and 78, respectively). RT-PCR analyses of spinal cord revealed an sevenfold increase of 14-3-3 γ expression in the spinal cord during EAE, particularly at later time points (* p < 0.05 at day 78 of EAE; Fig. 1a).Fig. 1


Role of glial 14-3-3 gamma protein in autoimmune demyelination.

Lee DH, Steinacker P, Seubert S, Turnescu T, Melms A, Manzel A, Otto M, Linker RA - J Neuroinflammation (2015)

Expression of 14-3-3 γ in glial cells. a RT-PCR analyses of spinal cord for the presence of 14-3-3 γ mRNA (n = 4 mice per time point). The relative expression in the spinal cord of naïve mice is set to 1. Note the increase of 14-3-3 γ expression in the spinal cord during EAE, particularly at later time points (*p < 0.05 at day 78 of EAE). b RT-PCR analysis of 14-3-3 γ mRNA in glial cells as compared to naïve spinal cord (n = 4 for spinal cord versus n = 9 independent cell preparations for astrocytes and n = 3 independent preparations for OPC). Note the significant 14-3-3 γ mRNA expression in glial cells as compared to total spinal cord (**p < 0.01). c, d Representative confocal laser scanning images after immunolabeling for 14-3-3 γ with GFAP in primary astrocyte culture c or Olig2 in primary OPC culture d shows expression of 14-3-3 γ on the protein level in the cytoplasm of some, but not all cells (arrows depict double labelled cells, bar = 20 μm)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4595275&req=5

Fig1: Expression of 14-3-3 γ in glial cells. a RT-PCR analyses of spinal cord for the presence of 14-3-3 γ mRNA (n = 4 mice per time point). The relative expression in the spinal cord of naïve mice is set to 1. Note the increase of 14-3-3 γ expression in the spinal cord during EAE, particularly at later time points (*p < 0.05 at day 78 of EAE). b RT-PCR analysis of 14-3-3 γ mRNA in glial cells as compared to naïve spinal cord (n = 4 for spinal cord versus n = 9 independent cell preparations for astrocytes and n = 3 independent preparations for OPC). Note the significant 14-3-3 γ mRNA expression in glial cells as compared to total spinal cord (**p < 0.01). c, d Representative confocal laser scanning images after immunolabeling for 14-3-3 γ with GFAP in primary astrocyte culture c or Olig2 in primary OPC culture d shows expression of 14-3-3 γ on the protein level in the cytoplasm of some, but not all cells (arrows depict double labelled cells, bar = 20 μm)
Mentions: First, we were interested whether 14-3-3 γ is expressed in EAE. To this end, we compared 14-3-3 γ expression in naïve spinal cord to inflamed spinal cord at different time points of EAE (maximum of disease, day 18 or chronic phase of EAE, day 50 and 78, respectively). RT-PCR analyses of spinal cord revealed an sevenfold increase of 14-3-3 γ expression in the spinal cord during EAE, particularly at later time points (* p < 0.05 at day 78 of EAE; Fig. 1a).Fig. 1

Bottom Line: At the same time, deficiency in 14-3-3 γ protein did not influence the immune response.Further histological studies revealed an increased susceptibility towards apoptosis in 14-3-3 γ-deficient OL in the inflamed spinal cord.These data argue for a pivotal role of 14-3-3 γ-mediated signalling pathways for OL protection in neuroinflammation.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Friedrich-Alexander-University Erlangen-Nuremberg, Schwabachanlage 6, D-91054, Erlangen, Germany.

ABSTRACT

Background: The family of 14-3-3 proteins plays an important role in the regulation of cell survival and death. Here, we investigate the role of the 14-3-3 gamma (14-3-3 γ) subunit for glial responses in autoimmune demyelination.

Methods: Expression of 14-3-3 γ in glial cell culture was investigated by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry. 14-3-3 γ knockout mice were subjected to murine myelin oligodendrocyte-induced experimental autoimmune encephalomyelitis (MOG-EAE), an animal model mimicking inflammatory features and neurodegenerative aspects of multiple sclerosis (MS).

Results: Expression studies in cell culture confined expression of 14-3-3 γ to both, oligodendrocytes (OL) and astrocytes. RT-PCR analysis revealed an increased expression of 14-3-3 γ mRNA in the spinal cord during the late chronic phase of MOG-EAE. At that stage, EAE was more severe in 14-3-3 γ knockout mice as compared to age- and gender-matched controls. Histopathological analyses on day 56 post immunization (p.i.) revealed significantly enhanced myelin damage as well as OL injury and secondary, an increase in axonal injury and gliosis in 14-3-3 γ -/- mice. At the same time, deficiency in 14-3-3 γ protein did not influence the immune response. Further histological studies revealed an increased susceptibility towards apoptosis in 14-3-3 γ-deficient OL in the inflamed spinal cord.

Conclusion: These data argue for a pivotal role of 14-3-3 γ-mediated signalling pathways for OL protection in neuroinflammation.

No MeSH data available.


Related in: MedlinePlus