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Published on: April 15, 2016
Measurement of NMDA receptor protein subunits in discrete hippocampal regions of kindled animals
1Department of Neurobiology, Duke University Medical Center, 401 Bryan Research Building, Durham, NC, 27710, USA.
Abstract:
Kindling refers to a phenomenon in which repeated application of initially subconvulsive electrical stimulations produces limbic and clonic motor seizures of progressively increasing severity. Once established, the increased excitability is lifelong. A diversity of studies demonstrate that kindling results in long lasting (28 days) alterations of the functional and pharmacologic properties of NMDA receptors, indicating that kindling may cause changes intrinsic to the NMDA receptor itself. Our previous studies disclosed no differences in NMDA receptor subunit gene or splice isoform mRNA expression between control and kindled animals 28 days after the last kindled seizure. Here, we extend those earlier studies by measuring levels of subunit protein for NMDAR1, NR2A, and NR2B in the hippocampus of control and kindled animals, 28 days after the last kindled seizure. We report that kindling does not effect long-lasting changes in the levels of NMDA receptor subunit protein. Together these findings support the idea that alterations in NMDA receptor protein expression do not contribute to the novel properties of NMDA receptors induced by kindling.
Insights
Kindling, a process causing lifelong brain hyperexcitability, does not alter NMDA receptor protein levels long-term. This suggests changes in NMDA receptor function, not expression, underlie kindling-induced brain alterations.
Area of Science:
- Neuroscience
- Epilepsy Research
- Molecular Biology
Background:
- Kindling is a model for epilepsy characterized by progressively severe seizures.
- Kindling induces long-lasting changes in NMDA receptor function.
- Previous studies found no changes in NMDA receptor mRNA expression post-kindling.
Purpose of the Study:
- To investigate if kindling affects NMDA receptor subunit protein levels.
- To determine if protein expression changes contribute to altered NMDA receptor function after kindling.
Main Methods:
- Kindling model in animals.
- Western blot analysis of hippocampal NMDA receptor subunits (NMDAR1, NR2A, NR2B).
- Comparison of protein levels between control and kindled animals 28 days post-seizure.
Main Results:
- No significant long-lasting changes in NMDAR1, NR2A, or NR2B protein levels were observed in kindled animals.
- These findings were consistent 28 days after the last seizure.
Conclusions:
- Kindling does not induce persistent alterations in the expression levels of key NMDA receptor subunits.
- Altered NMDA receptor protein expression is unlikely to be the mechanism behind kindling-induced functional changes.

