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减少常见髓鞘 cDNA 序列后对鼠脊髓转录组的分析。

Murine spinal cord transcriptome analysis following reduction of prevalent myelin cDNA sequences.

机构信息

Solomon Park Research Institute, Kirkland, WA 98034, USA.

出版信息

Cell Mol Neurobiol. 2009 Dec;29(8):1109-19. doi: 10.1007/s10571-009-9472-5.

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.

摘要

从通过抑制性消减杂交从鼠脊髓减去白质的 cDNA 文库中随机选择的 1000 个克隆中,通过点印迹分析鉴定出 220 个差异表达的克隆。通过 BLAST 程序进行的序列分析鉴定出 140 个独特的基因。(1) 与以前使用视皮层作为驱动的类似消减相比,来自髓磷脂和其他神经胶质来源的已知序列的百分比降低了约 75%。(2) 差异表达的基因倾向于反映脊髓结构和功能的现有预期。(3) 在本研究中,脊髓中差异表达的所有基因的约 35%也已知在 UniGene 数据库中为该结构差异表达。(4) 在本研究中差异表达的所有基因的约 33%在根据 UniGene 数据库测量脊髓时记录为不存在,表明目前的技术没有记录该结构中约三分之一的差异表达基因。(5) 所有差异表达基因的约 15%是未知的、假定的或假设的蛋白质产物。(6) 所有差异表达基因的约 4%是用于小鼠的新表达序列标签。本研究表明,在比较大脑区域时,减少与神经胶质相关序列的存在非常重要。结论是,当使用白质作为驱动时,脊髓中存在一些髓磷脂序列表明,与白质和胼胝体代表的区域相比,该结构中的髓鞘形成更加活跃。

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