Wise Young
10-01-2001, 11:29 PM
Journal of Neurochemistry, Vol. 76, No. 3, 2001 797-805
© 2001 International Society for Neurochemistry
Expression of splice variants of the NR1 subunit of the N-methyl-D-aspartate receptor in the normal and injured rat spinal cord
K. L. Prybylowski*, S. D. Grossman, J. R. Wrathall and B. B. Wolfe*
* Department of Pharmacology, Georgetown University, Washington DC, USA
Department of Cell Biology, Georgetown University, Washington DC, USA
Department of Neuroscience, Georgetown University, Washington DC, USA
Address correspondence and reprint requests to Dr Barry Wolfe, Department of Pharmacology, 3900 Reservoir Road, Washington DC 20007, USA. E-mail: bwolfe01@gusun.georgetown.edu
Quantitative western blot analysis in laminectomy control spinal cords of adult rats was used to provide the first report of the normal expression patterns of the N1, C1, C2 and C2' cassettes in the cervical, thoracic and lumbar regions of the spinal cord as a percent of total NR1 subunit protein. In all regions studied, the C1 and C2 cassettes were usually contained in less than 10% of total NR1 protein. In contrast, approximately 90% of total NR1 protein contained the C2' cassette. A significant proportion of total NR1 protein (approximately 30%) also contained the N1 cassette. These data are consistent with expression of NR1000 (NR1-4a) and NR1100 (NR1-4b) as the dominant splice forms in the spinal cord. Splice variant expression was also studied following incomplete, contusive spinal cord injury (SCI) to the thoracic level 8 (T8) region. This injury did not change expression of the C1 or C2 cassette in any region of the spinal cord acutely at 24 h or chronically at 1 month. There was an increase in expression of the N1 cassette in the lumbar regions 1 month after injury (p < 0.05). These data indicate that SCI induces distal changes in NR1 splice variant expression, which may play a role in the adaptive response of neurons in the chronically injured spinal cord.
Key Words: glutamate receptor - quantitative western blot - spinal cord injury
Abbreviations used: PKC, protein kinase C; PVDF, polyvinylidene difluoride; SCI, spinal cord injury; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, T-thoracic level
© 2001 International Society for Neurochemistry
Expression of splice variants of the NR1 subunit of the N-methyl-D-aspartate receptor in the normal and injured rat spinal cord
K. L. Prybylowski*, S. D. Grossman, J. R. Wrathall and B. B. Wolfe*
* Department of Pharmacology, Georgetown University, Washington DC, USA
Department of Cell Biology, Georgetown University, Washington DC, USA
Department of Neuroscience, Georgetown University, Washington DC, USA
Address correspondence and reprint requests to Dr Barry Wolfe, Department of Pharmacology, 3900 Reservoir Road, Washington DC 20007, USA. E-mail: bwolfe01@gusun.georgetown.edu
Quantitative western blot analysis in laminectomy control spinal cords of adult rats was used to provide the first report of the normal expression patterns of the N1, C1, C2 and C2' cassettes in the cervical, thoracic and lumbar regions of the spinal cord as a percent of total NR1 subunit protein. In all regions studied, the C1 and C2 cassettes were usually contained in less than 10% of total NR1 protein. In contrast, approximately 90% of total NR1 protein contained the C2' cassette. A significant proportion of total NR1 protein (approximately 30%) also contained the N1 cassette. These data are consistent with expression of NR1000 (NR1-4a) and NR1100 (NR1-4b) as the dominant splice forms in the spinal cord. Splice variant expression was also studied following incomplete, contusive spinal cord injury (SCI) to the thoracic level 8 (T8) region. This injury did not change expression of the C1 or C2 cassette in any region of the spinal cord acutely at 24 h or chronically at 1 month. There was an increase in expression of the N1 cassette in the lumbar regions 1 month after injury (p < 0.05). These data indicate that SCI induces distal changes in NR1 splice variant expression, which may play a role in the adaptive response of neurons in the chronically injured spinal cord.
Key Words: glutamate receptor - quantitative western blot - spinal cord injury
Abbreviations used: PKC, protein kinase C; PVDF, polyvinylidene difluoride; SCI, spinal cord injury; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, T-thoracic level