Developmental expression. compartmentalization, and possible role in excitotoxicity of a putative NMDA receptor protein in cultured hippocampal-neurons
Abstract
The mechanisms regulating the expression and localization of excitatory amino acid (EAA) neurotransmitter receptors in neurons of the developing mammalian brain, and roles for these receptors in the plasticity and degeneration of neural circuits are not well understood. We previously isolated and characterized a 71 kDa glutamate binding protein (GBP) from rat brain, and have recently obtained evidence that this GBP is a component of a functional N-methyl-D-aspartate (NMDA) receptor-ion channel complex. We have now used antibodies to this putative NMDA receptor protein to examine its expression and localization, and consequences of its activation in cultured embryonic (18 day) rat hippocampal neurons. Immunocytochemistry and Western blots using monoclonal antibodies to the GBP demonstrated an increase in GBP-positive neurons and their staining intensity with time in culture. GBP was localized to the somata and dendrites of pyramidal-like neurons and was sparse or absent in the axons. The expression and compartmentalization of GBP occurred in isolated neurons indicating that direct cell interactions were not required for these processes. Cell surface staining for GBP occurred in patches on the soma and dendrites. The developmental expression of GBP immunoreactivity closely paralleled the expression of sensitivity to NMDA neurotoxicity. There was a direct relationship between GBP immunoreactivity and neuronal vulnerability to glutamate-induced degeneration; vulnerable neurons st…
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Cultures that showed a purity of at least 90% were grown further until 14 DIV.
“Those cultures that showed a purity of at least 90% were grown further until 14 DIV when neurons reach their mature form and express functional glutamatergic receptors.25”
From Blocking the Interaction between Apolipoprotein E and Aβ Reduces Intraneuronal Accumulation of Aβ and Inhibits Synaptic Degeneration (2013) · cites this paper as [25]
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Glutamate in the Pathogenesis of Gliomas
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Glutamate in the Pathogenesis of Gliomas
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Glutamate in the Pathogenesis of Gliomas
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- Authors
- Mattson, M. P., Wang, H., & Michaelis, E. K.
- Affiliations
- University of Kentucky (US)
- University of Kansas (US)
- Corresponding author
- Mattson, Mark P.
- Venue
- Brain Research
- Publisher
- Elsevier BV
- Published
- 1991-11
- Volume
- 565
- Issue
- 1
- Pages
- 94-108
- ISSN
- 0006-8993 (ISSN-L)
- OpenAlex
- W2023298754
- Cited by
- 98
- References
- 48
- Citations by year
- 2022: 2 · 2020: 1 · 2019: 1 · 2015: 2 · 2014: 1 · 2013: 3 · 2012: 2
- FWCI
- 3.27
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Brain Research (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r01ns029001-07)
- National Institute on Alcohol Abuse and Alcoholism (AA 04732)
- National Institute on Aging (P50AG005144, AG-05144)
- National Institute of Neurological Disorders and Stroke (NS29001)
- Topics
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Amino Acid Enzymes and Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nicotinic Acetylcholine Receptors Study
- Keywords
- NMDA receptor, excitotoxicity, hippocampal neurons, embryonic development, subcellular localization, dendrites, pyramidal neurons, neuronal vulnerability, NMDA neurotoxicity, kainate neurotoxicity, glutamate receptor expression
- MeSH terms
- Animals, Antibodies, Antibodies, Monoclonal, Cell Membrane, Cell Survival, Cells, Cultured, Electrophoresis, Polyacrylamide Gel, Embryo, Mammalian, Hippocampus, Immunohistochemistry, Ion Channels, Kinetics, Molecular Weight, Neurons, Pyramidal Tracts, Rats, Inbred Strains, Time Factors, Blotting, Western, Receptors, N-Methyl-D-Aspartate, N-Methylaspartate, Rats
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