Profiling of gene expression related neuronal death by intracellular Aβ1-42

细胞内 Aβ1-42 相关神经元死亡的基因表达谱分析

• 批准号: 16500233
• 负责人: UCHIDA Yoko
• 金额: $ 2.5万
• 依托单位: Tokyo Metropolitan Institute of Gerontology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16500233/
• 关键词: β-amyloid; intracellular Aβ; neuronal death; microarray; AICD; 細胞内Ass; 細胞死関連遺伝子

The purpose of this study is to clarify the molecular mechanisms of neuronal death induced by intracellular Aβ by using gene expression profiling. Results are as follows. (1) First we determined the effect of intracellular Aβ on neuronal death by using transfection experiments of APP-CTFβ(C99)-myc-IRES-hrGFP cDNA into cultured cortical neurons. C99 expressed in neurons really induced cell death: however different detection procedures for C99-expressing cells resulted in different time course and different degree of neuronal death. Larger proportion of dead C99-expressing neurons were detected by anti-myc antibodies than that by hrGFP fluorescence, indicating that intracellular deposition of C99 may induce neuronal death. (2) Then we analyzed the gene expression profiles related to cell death caused by the accumulation of C99 by using microarray technology. Forty-six genes were identified as the genes altered their expressions by the intracellular accumulation of C99. To confirm the altered expressions of 10 genes, we performed QRT-PCR by comparing the gene expressions in myc-expressing or △126MAP1B-expressing cells, which did not show cell death. Although there was no significant difference in gene expressions between C99-expressing cell and myc-expressing cells, the expressions of several genes were altered in C99-expressing cell compared with △126MAP1B-expressing cells. (3) Now we are examining these gene expressions in C99-expressing cell compared with APP-CTF alpha or with C99-D644A by using QRT-PCR.

这项研究的目的是通过使用基因表达分析来阐明由细胞内Aβ诱导的神经元死亡的分子机制。结果如下。 （1）首先，我们通过使用APP-CTFβ（C99）-MYC-IRES-HRGFP cDNA对培养的皮质神经元的转化实验来确定细胞内Aβ对神经元死亡的影响。在神经元中表达的C99确实诱导了细胞死亡：但是表达C99细胞的不同检测程序导致不同的时间过程和不同程度的神经元死亡。与HRGFP荧光相比，通过抗MYC抗体检测到的表达C99神经元的比例更大，这表明C99的细胞内沉积可能诱导神经元死亡。 （2）然后，我们分析了使用微阵列技术的C99积累引起的与细胞死亡有关的基因表达谱。四十六个基因被鉴定为基因通过C99的细胞内积累改变其表达。为了确认10个基因的表达变化，我们通过比较表达MYC表达或表达MYC 126MAP1B的细胞中的基因表达来进行QRT-PCR，该细胞没有显示细胞死亡。尽管表达C99的细胞和表达MYC的细胞之间的基因表达没有显着差异，但与△126MAP1B的细胞相比，表达C99的细胞中几种基因的表达发生了变化。 （3）现在，我们使用QRT-PCR检查了与APP-CTF alpha或C99-D644A相比，我们正在检查表达C99的细胞中的这些基因表达。