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Phosphorylated TDP-43 in frontotemporal lobar degeneration and amyotrophic lateral sclerosis
- Source :
- Annals of Neurology. 64:60-70
- Publication Year :
- 2008
- Publisher :
- Wiley, 2008.
-
Abstract
- Objective TAR DNA-binding protein of 43kDa (TDP-43) is deposited as cytoplasmic and intranuclear inclusions in brains of patients with frontotemporal lobar degeneration with ubiquitinated inclusions (FTLD-U) and amyotrophic lateral sclerosis (ALS). Previous studies reported that abnormal phosphorylation takes place in deposited TDP-43. The aim of this study was to identify the phosphorylation sites and responsible kinases, and to clarify the pathological significance of phosphorylation of TDP-43. Methods We generated multiple antibodies specific to phosphorylated TDP-43 by immunizing phosphopeptides of TDP-43, and analyzed FTLD-U and ALS brains by immunohistochemistry, immunoelectron microscopy, and immunoblots. In addition, we performed investigations aimed at identifying the responsible kinases, and we assessed the effects of phosphorylation on TDP-43 oligomerization and fibrillization. Results We identified multiple phosphorylation sites in carboxyl-terminal regions of deposited TDP-43. Phosphorylation-specific antibodies stained more inclusions than antibodies to ubiquitin and, unlike existing commercially available anti–TDP-43 antibodies, did not stain normal nuclei. Ultrastructurally, these antibodies labeled abnormal fibers of 15nm diameter and on immunoblots recognized hyperphosphorylated TDP-43 at 45kDa, with additional 18 to 26kDa fragments in sarkosyl-insoluble fractions from FTLD-U and ALS brains. The phosphorylated epitopes were generated by casein kinase-1 and -2, and phosphorylation led to increased oligomerization and fibrillization of TDP-43. Interpretation These results suggest that phosphorylated TDP-43 is a major component of the inclusions, and that abnormal phosphorylation of TDP-43 is a critical step in the pathogenesis of FTLD-U and ALS. Phosphorylation-specific antibodies will be powerful tools for the investigation of these disorders. Ann Neurol 2008
- Subjects :
- Male
Pathology
medicine.medical_specialty
Immunoelectron microscopy
macromolecular substances
Biology
Antibodies
Article
Epitope
Inclusion bodies
Epitopes
Antibody Specificity
mental disorders
medicine
Humans
Phosphorylation
Amyotrophic lateral sclerosis
Aged
Aged, 80 and over
Inclusion Bodies
Casein Kinase I
Kinase
Amyotrophic Lateral Sclerosis
Brain
nutritional and metabolic diseases
Neurofibrillary Tangles
Frontotemporal lobar degeneration
Middle Aged
medicine.disease
Molecular biology
Peptide Fragments
Protein Structure, Tertiary
nervous system diseases
DNA-Binding Proteins
Neurology
bacteria
Immunohistochemistry
Dementia
Female
Neurology (clinical)
Subjects
Details
- ISSN :
- 15318249 and 03645134
- Volume :
- 64
- Database :
- OpenAIRE
- Journal :
- Annals of Neurology
- Accession number :
- edsair.doi.dedup.....b22033ae5fecae1d3a57ac1a342fa0f1