cDNA微阵列技术比较胚胎大鼠脑和脊髓神经干细胞的基因差异表达  

COMPARISON OF GENES DIFFERENTIAL EXPRESSION BETWEEN NEURAL STEM CELLS DERIVED FROM EMBRYONIC RAT BRAIN AND SPINAL CORD USING cDNA MICROARRAY

作　　者：胡建国[1] 富赛里[1] 李莹[1] 尹岚[1] 金建强[1] 沈天伟[2] 陆佩华[1] 徐晓明[1] 

机构地区：[1]上海第二医科大学神经生物学实验室,上海200025 [2]上海第二医科大学免疫学研究所,上海200025

出　　处：《解剖学报》2005年第2期145-149,共5页Acta Anatomica Sinica

基　　金：国家重点基础研究发展计划(973)项目(2003CB515302);上海市科技发展基金(00JC14021)

摘　　要：目的　从分子水平探讨脑和脊髓来源的神经干细胞(NSCs)的生物学特性,并鉴定这两种来源的NSCs 的分子表达差异。　方法　应用cDNA微阵列技术对脑和脊髓来源的NSCs的基因表达情况进行检测和比较,在成 功分离、培养和鉴定脑和脊髓两种来源的NSCs的基础上,抽提细胞总RNA并纯化、定量,以32P逆转录标记合成 cDNA探针。AltasTMcDNAExpressionArray(ClontechCo)与探针杂交、洗膜后在磷屏上曝光并扫描成像,以Arrayvision 5.1软件分析杂交结果。　结果　cDNA阵列膜上覆盖的1176个基因中,绝大部分基因表达无明显差异,但也有14 个基因在两种来源的NSCs中的表达有显著差异,其中8个在脑来源的NSCs中表达较高,6个在脊髓来源的NSCs 中表达较显著。在两种来源的NSCs都明显表达的基因中,许多分子如P 选择素(P selectin)、钙黏素(cadherin)、乙酰 胆碱受体α、cyclins、某些转录因子和原癌基因等可能在NSCs的自我更新(增殖)、神经球形成和保持不分化状态中 起重要作用。　结论　两种来源的NSCs的生物学特性基本相同,但也存在一定的差异。Objective To compare differential gene expression between NSCs derived from brain and spinal cord and investigate biological feature of NSCs and difference between brain-derived NSCs and spinal cord-derived NSCs. Methods Brain-and spinal cord-derived stem cells(NSCs) were obtained from E16 rats and cultured in vitro in the presence of fibroblast growth factor-2(FGF-2) and epidermal growth factor(EGF). Total RNA of these cells was isolated, purified and quantified. Five μg of the total RNA was used to synthesize[α- 32P] dATP-radio-labelled cDNA probes by reverse transcriptase. Rat 1.2 Atlas cDNA expession arrays containing 1?176 genes (CLONTECH) were hybridized with cDNA probes. The arrays were washed and exposed to a phosphorimaging screen. The hybridization signals were scanned with a phosphorimager and analyzed with Arrayvision 5.1. Results Among 1?176 genes examined, 14 genes showed significant difference between the brain-and spinal cord-derived NSCs, while over 200 genes expressed equally between the two populations using a 2-fold change in signal intensity and P<0.05 criteria. Among genes expressed differentially, 8 genes were significantly higher and 6 genes were significantly lower in brain-derived NSCs than in spinal cord-derived NSCs. Among genes that showed same expressional changes between brain and spinal cord-derived NSCs, many molecules such as P-selectin, Cadherin, acetylcholine receptor alpha, cyclins, some transcription factors and oncogenes may play a role in self-renewal, proliferation and maintainance of NSCs.Conclusion Our findings suggest that patters of gene expression in the brain-and spinal cord-derived NSCs are similar. However, differences between the two also exist.

关 键 词：基因表达 神经干细胞 脑 脊髓 CDNA微阵列 大鼠 

分 类 号：Q512[生物学—生物化学]