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Neurodegeneration in Autoimmune Optic Neuritis Is Associated with Altered APP Cleavage in Neurons and Up-Regulation of p53.

Herold S, Kumar P, Wichert SP, Kretzschmar B, Bähr M, Rossner MJ, Hein K - PLoS ONE (2015)

Bottom Line: Furthermore, genes that are highly specific for autoimmune-driven neurodegeneration were compared to those regulated in RGCs after optic nerve axotomy at corresponding time points.Furthermore, using ingenuity pathway analysis (IPA), we identified amyloid precursor protein (APP) as a potential upstream regulator of changes in gene expression in the preclinical EAE but neither in clinical EAE, nor at any time point after optic nerve transection.Therefore, the gene pathway analysis lead to the hypothesis that altered cleavage of APP in neurons in the preclinical phase of EAE leads to the enhanced production of APP intracellular domain (AICD), which in turn acts as a transcriptional regulator and thereby initiates an apoptotic signaling cascade via up-regulation of the target gene p53.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, University Medicine Göttingen, Göttingen, Germany.

ABSTRACT
Multiple Sclerosis (MS) is a chronic autoimmune inflammatory disease of the central nervous system (CNS). Histopathological and radiological analysis revealed that neurodegeneration occurs early in the disease course. However, the pathological mechanisms involved in neurodegeneration are poorly understood. Myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis (EAE) in Brown Norway rats (BN-rats) is a well-established animal model, especially of the neurodegenerative aspects of MS. Previous studies in this animal model indicated that loss of retinal ganglion cells (RGCs), the neurons that form the axons of the optic nerve, occurs in the preclinical phase of the disease and is in part independent of overt histopathological changes of the optic nerve. Therefore, the aim of this study was to identify genes which are involved in neuronal cell loss at different disease stages of EAE. Furthermore, genes that are highly specific for autoimmune-driven neurodegeneration were compared to those regulated in RGCs after optic nerve axotomy at corresponding time points. Using laser capture micro dissection we isolated RNA from unfixed RGCs and performed global transcriptome analysis of retinal neurons. In total, we detected 582 genes sequentially expressed in the preclinical phase and 1150 genes in the clinical manifest EAE (P < 0.05, fold-induction >1.5). Furthermore, using ingenuity pathway analysis (IPA), we identified amyloid precursor protein (APP) as a potential upstream regulator of changes in gene expression in the preclinical EAE but neither in clinical EAE, nor at any time point after optic nerve transection. Therefore, the gene pathway analysis lead to the hypothesis that altered cleavage of APP in neurons in the preclinical phase of EAE leads to the enhanced production of APP intracellular domain (AICD), which in turn acts as a transcriptional regulator and thereby initiates an apoptotic signaling cascade via up-regulation of the target gene p53.

No MeSH data available.


Related in: MedlinePlus

Workflow for data analysis.After sacrificing the animals, eyes were enucleated from the animals immunized with myelin oligodendrocyte glycoprotein (MOG) (A) and from the group of animals with optic nerve transection (ONT) (B), at early (day 7 post MOG-immunization (MOG_7dpi) or day 7 after ONT (ONT_7)) and late time points (day 1 of experimental autoimmune encephalomyelitis (d1EAE) or day 14 after ONT (ONT_14)). Retinal ganglion cells (RGCs) were isolated using laser capture microdissection (LCM) and a global transcriptome analysis was performed. Selection of relevant pathways and network specific for MOG-EAE was detected using Ingenuity Pathway Analysis (IPA).
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pone.0138852.g001: Workflow for data analysis.After sacrificing the animals, eyes were enucleated from the animals immunized with myelin oligodendrocyte glycoprotein (MOG) (A) and from the group of animals with optic nerve transection (ONT) (B), at early (day 7 post MOG-immunization (MOG_7dpi) or day 7 after ONT (ONT_7)) and late time points (day 1 of experimental autoimmune encephalomyelitis (d1EAE) or day 14 after ONT (ONT_14)). Retinal ganglion cells (RGCs) were isolated using laser capture microdissection (LCM) and a global transcriptome analysis was performed. Selection of relevant pathways and network specific for MOG-EAE was detected using Ingenuity Pathway Analysis (IPA).

Mentions: We used 6 different experimental groups of animals which were as follow: 1. MOG-immunized animals analyzed at day 7 post MOG-immunization (MOG_7dpi) or 2. At the day of clinical manifestation of the disease (d1EAE). 3. Sham-immunized control group sacrificed at day 7 post immunization or 4. at day 14 post immunization (Sham). 5. Animals with optic nerve transection (ONT) analyzed at day 7 after axotomy (ONT_7) or 6. At day 14 after axotomy (ONT_14) (Fig 1). 6 eyes per group were used for the microarray and Western blots analysis, 8 eyes were used for quantitative real-time PCR (qPCR) analysis and 3 eyes for immunohistochemistry. Animals were sacrificed using an overdose of carbon dioxide.


Neurodegeneration in Autoimmune Optic Neuritis Is Associated with Altered APP Cleavage in Neurons and Up-Regulation of p53.

Herold S, Kumar P, Wichert SP, Kretzschmar B, Bähr M, Rossner MJ, Hein K - PLoS ONE (2015)

Workflow for data analysis.After sacrificing the animals, eyes were enucleated from the animals immunized with myelin oligodendrocyte glycoprotein (MOG) (A) and from the group of animals with optic nerve transection (ONT) (B), at early (day 7 post MOG-immunization (MOG_7dpi) or day 7 after ONT (ONT_7)) and late time points (day 1 of experimental autoimmune encephalomyelitis (d1EAE) or day 14 after ONT (ONT_14)). Retinal ganglion cells (RGCs) were isolated using laser capture microdissection (LCM) and a global transcriptome analysis was performed. Selection of relevant pathways and network specific for MOG-EAE was detected using Ingenuity Pathway Analysis (IPA).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591258&req=5

pone.0138852.g001: Workflow for data analysis.After sacrificing the animals, eyes were enucleated from the animals immunized with myelin oligodendrocyte glycoprotein (MOG) (A) and from the group of animals with optic nerve transection (ONT) (B), at early (day 7 post MOG-immunization (MOG_7dpi) or day 7 after ONT (ONT_7)) and late time points (day 1 of experimental autoimmune encephalomyelitis (d1EAE) or day 14 after ONT (ONT_14)). Retinal ganglion cells (RGCs) were isolated using laser capture microdissection (LCM) and a global transcriptome analysis was performed. Selection of relevant pathways and network specific for MOG-EAE was detected using Ingenuity Pathway Analysis (IPA).
Mentions: We used 6 different experimental groups of animals which were as follow: 1. MOG-immunized animals analyzed at day 7 post MOG-immunization (MOG_7dpi) or 2. At the day of clinical manifestation of the disease (d1EAE). 3. Sham-immunized control group sacrificed at day 7 post immunization or 4. at day 14 post immunization (Sham). 5. Animals with optic nerve transection (ONT) analyzed at day 7 after axotomy (ONT_7) or 6. At day 14 after axotomy (ONT_14) (Fig 1). 6 eyes per group were used for the microarray and Western blots analysis, 8 eyes were used for quantitative real-time PCR (qPCR) analysis and 3 eyes for immunohistochemistry. Animals were sacrificed using an overdose of carbon dioxide.

Bottom Line: Furthermore, genes that are highly specific for autoimmune-driven neurodegeneration were compared to those regulated in RGCs after optic nerve axotomy at corresponding time points.Furthermore, using ingenuity pathway analysis (IPA), we identified amyloid precursor protein (APP) as a potential upstream regulator of changes in gene expression in the preclinical EAE but neither in clinical EAE, nor at any time point after optic nerve transection.Therefore, the gene pathway analysis lead to the hypothesis that altered cleavage of APP in neurons in the preclinical phase of EAE leads to the enhanced production of APP intracellular domain (AICD), which in turn acts as a transcriptional regulator and thereby initiates an apoptotic signaling cascade via up-regulation of the target gene p53.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, University Medicine Göttingen, Göttingen, Germany.

ABSTRACT
Multiple Sclerosis (MS) is a chronic autoimmune inflammatory disease of the central nervous system (CNS). Histopathological and radiological analysis revealed that neurodegeneration occurs early in the disease course. However, the pathological mechanisms involved in neurodegeneration are poorly understood. Myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis (EAE) in Brown Norway rats (BN-rats) is a well-established animal model, especially of the neurodegenerative aspects of MS. Previous studies in this animal model indicated that loss of retinal ganglion cells (RGCs), the neurons that form the axons of the optic nerve, occurs in the preclinical phase of the disease and is in part independent of overt histopathological changes of the optic nerve. Therefore, the aim of this study was to identify genes which are involved in neuronal cell loss at different disease stages of EAE. Furthermore, genes that are highly specific for autoimmune-driven neurodegeneration were compared to those regulated in RGCs after optic nerve axotomy at corresponding time points. Using laser capture micro dissection we isolated RNA from unfixed RGCs and performed global transcriptome analysis of retinal neurons. In total, we detected 582 genes sequentially expressed in the preclinical phase and 1150 genes in the clinical manifest EAE (P < 0.05, fold-induction >1.5). Furthermore, using ingenuity pathway analysis (IPA), we identified amyloid precursor protein (APP) as a potential upstream regulator of changes in gene expression in the preclinical EAE but neither in clinical EAE, nor at any time point after optic nerve transection. Therefore, the gene pathway analysis lead to the hypothesis that altered cleavage of APP in neurons in the preclinical phase of EAE leads to the enhanced production of APP intracellular domain (AICD), which in turn acts as a transcriptional regulator and thereby initiates an apoptotic signaling cascade via up-regulation of the target gene p53.

No MeSH data available.


Related in: MedlinePlus