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Semi-quantitative analysis of alpha-synuclein in subcellular pools of rat brain neurons: an immunogold electron microscopic study using a C-terminal specific monoclonal antibody.
Semi-quantitative analysis of alpha-synuclein in subcellular pools of rat brain neurons: an immunogold electron microscopic study using a C-terminal specific monoclonal antibody.
- Source :
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Brain research [Brain Res] 2008 Dec 09; Vol. 1244, pp. 40-52. Date of Electronic Publication: 2008 Sep 04. - Publication Year :
- 2008
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Abstract
- Alpha-synuclein (alpha-Syn) is a brain-enriched protein of 140 amino acids. Despite of strong evidence showing the implication of the protein in the pathogenesis of several neurodegenerative diseases, its physiological function remains poorly understood. To study the physiological function of alpha-Syn, a depiction of its precise subcellular localization is necessary. Although alpha-Syn expression in the brain has been extensively investigated using several different antibodies, its precise subcellular localization in neurons remains elusive. In this study, immunogold electron microscopy with a newly produced 3D5 monoclonal antibody recognizing the C-terminal 115-121 amino acids of alpha-Syn was used to examine its subcellular localization in rat brain neurons. In addition, the relative amount of the protein in different subcellular pools of the neurons in several brain regions was evaluated and compared. The results showed that alpha-Syn-positive gold particles were unevenly distributed in axons, presynaptic terminals, cytoplasm and nucleus in the neuron, with the density of gold particles being greater in presynaptic terminals and nucleus than in other subcellular pools. In the cytoplasmic region, relatively dense gold particles were seen in some mitochondria. In the same subcellular pools, the density of gold particles was varied among the neurons from different brain regions. Although the cortical neurons showed much higher density of gold particles in the presynaptic terminals and nuclei than in striatal, hippocampal and substantia nigral neurons, the density of gold particles in their mitochondria was much lower compared with the mitochondria of striatal, hippocampal and substantia nigral neurons. The relative high level of mitochondrial alpha-Syn in hippocampus, striatum and substantia nigral neurons may have special pathophysiological significance, which deserves further investigation.
- Subjects :
- Analysis of Variance
Animals
Antibodies, Monoclonal immunology
Antibodies, Monoclonal metabolism
Antibody Specificity immunology
Axons metabolism
Axons ultrastructure
Blotting, Western
Brain cytology
Cell Nucleus metabolism
Cell Nucleus ultrastructure
Cerebral Cortex cytology
Cerebral Cortex metabolism
Corpus Striatum cytology
Corpus Striatum metabolism
Cytoplasm metabolism
Cytoplasm ultrastructure
Hippocampus cytology
Hippocampus metabolism
Immunohistochemistry methods
Microscopy, Immunoelectron
Mitochondria metabolism
Mitochondria ultrastructure
Neurons ultrastructure
Presynaptic Terminals metabolism
Presynaptic Terminals ultrastructure
Rats
Rats, Sprague-Dawley
Substantia Nigra cytology
Substantia Nigra metabolism
alpha-Synuclein immunology
Brain metabolism
Neurons metabolism
alpha-Synuclein metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1872-6240
- Volume :
- 1244
- Database :
- MEDLINE
- Journal :
- Brain research
- Publication Type :
- Academic Journal
- Accession number :
- 18817762
- Full Text :
- https://doi.org/10.1016/j.brainres.2008.08.067