Murine Spinal Cord Transcriptome Analysis Following Reduction of Prevalent Myelin cDNA Sequences
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Abstract
From 1,000 randomly selected colonies from cDNA libraries derived from murine spinal cord subtracted against white matter by means of suppression subtractive hybridization, 220 clones were identified as differentially expressed by dot blot analysis. Sequence analysis by the BLAST programming identified 140 unique genes. (1) The percentage of known sequences from myelin and other glial sources was reduced by approximately 75% over previous, similar subtractions employing visual cortex as the driver. (2) Differentially expressed genes tended to reflect existing expectations concerning structure and function of the spinal cord. (3) About 35% of all genes differentially expressed in the spinal cord in this study are also known to be differentially expressed for this structure as tabulated in the UniGene database. (4) About 33% of all genes differentially expressed in the present study are recorded as not present when measured in the spinal cord according to the UniGene database indicating that present techniques are not recording about a third of differentially expressed genes in this structure. (5) About 15% of all differentially expressed genes are for unknown, putative or hypothetical protein products. (6) About 4% of all differentially expressed genes are novel expressed sequence tags for the mouse. The current study demonstrates the importance of reducing the presence of glial associated sequences when comparing brain regions. It is concluded that the persistence of some myelin sequences in the spinal cord when white matter is used as the driver indicates that myelination is more active in this structure than for those areas represented by white matter and corpus callosum.
Keywords
Spinal cord transcriptome Suppression subtractive hybridization Glia White matter Motor neuron diseaseNotes
Acknowledgments
This work is supported by a bequest from the estate of Frances and Wesley Johnson and the generous donations of numerous individuals to Solomon Park Research Institute. We would like to thank the members of the board of directors of Solomon Park Research Institute for their tireless dedication to the Institute and the ongoing research effort. We are especially appreciative of the many helpful remarks and suggestions from Dr. Susan A. Martinis concerning the role of leucyl-tRNA synthetase.
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