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

2006 Fiscal Year Final Research Report Summary

• Project/Area Number: 16500233
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Nerve anatomy/Neuropathology
• Research Institution: Tokyo Metropolitan Institute of Gerontology
• Principal Investigator: UCHIDA Yoko Tokyo Metropolitan Institute of Gerontology, Tokyo Metropolitan Institute of Gerontology, Senior Research Scientist (60133633)
• Co-Investigator(Kenkyū-buntansha): GOMI Fujiya Tokyo Metropolitan Foundation for Research on Aging and Promotion of Human, Research Scientist (40205620)
• Project Period (FY): 2004 – 2006
• Keywords: β-amyloid / intracellular Aβ / neuronal death / microarray / AICD

Research Abstract

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.