Shinya Tokuhiro

Caspase activation during apoptotic cell death induced by expanded polyglutamine in N2a cells.

Huntington disease (HD) is an autosomal dominant neurodegenerative disorder. To investigate the mechanism of neurodegeneration induced by mutant huntingtin, we developed a stable neuro2a cell line expressing truncated N-terminal huntingtin (tNhtt) with EGFP using the ecdysone-inducible system. The formation of aggregates and the cell death induced by expression of tNhtt with expanded polyglutamine was repeat length- and dose-dependent. Caspases were activated, and the death substrates of caspases, lamin B and ICAD (an inhibitor of caspase-activated DNase), were cleaved in this cell death process. The cleavage of lamin B was inhibited by caspase inhibitors. These findings suggest that the cell death induced by tNhtt with expanded polyglutamine is mediated by caspases.

Molecular Dissection of Domains in Mutant Presenilin 2 That Mediate Overproduction of Amyloidogenic Forms of Amyloid β Peptides INABILITY OF TRUNCATED FORMS OF PS2 WITH FAMILIAL ALZHEIMER’S DISEASE MUTATION TO INCREASE SECRETION OF Aβ42

Mutations in presenilin (PS) 1 or PS2 genes account for the majority of early-onset familial Alzheimer’s disease, and these mutations have been shown to increase production of species of amyloid β peptide (Aβ) ending at residue 42, i.e.the most amyloidogenic form of Aβ. To gain insight into the molecular mechanisms whereby mutant PS induces overproduction of Aβ42, we constructed cDNAs encoding mutant and/or truncated forms of PS2 and examined the secretion of Aβ42 from COS or neuro2a cells transfected with these genes. Cells expressing full-length PS2 harboring both N141I and M239V mutations in the same polypeptide induced overproduction of Aβ42, although the levels of Aβ42 were comparable with those in cells engineered to express PS2 with one or the other of these PS2 mutations. In contrast, cells engineered to express partially truncated PS2 (eliminating the COOH-terminal third of PS2 while retaining the endoproteolytic NH2-terminal fragment) and harboring a N141I mutation, as well as cells expressing COOH-terminal fragments of PS2, did not overproduce Aβ42, and the levels of Aβ42 were comparable with those in cells that expressed full-length, wild-type PS2 or fragments thereof. These data indicate that: (i) the Aβ42-promoting effects of mutant PS2 proteins reach the maximum level with a given single amino acid substitution (i.e. N141I or M239V); and (ii) the expression of full-length mutant PS2 is required for the overproduction of Aβ42. Hence, cooperative interactions of NH2- and COOH-terminal fragments generated from full-length mutant PS2 may be important for the overproduction of Aβ42 that may underlie familial Alzheimer’s disease.

β-Amyloid-specific upregulation of stearoyl coenzyme A desaturase-1 in macrophages

beta-Amyloid peptide (A beta), a major component of senile plaques, the formation of which is characteristic of Alzheimers disease (AD), is believed to induce inflammation of the brain mediated by microglia, leading to neuronal cell loss. In this study, we performed an oligonucleotide microarray analysis to investigate the molecular events underlying the A beta-induced activation of macrophages and its specific suppression by the A beta-specific-macrophage-activation inhibitor, RS-1178. Of the approximately 36,000 genes and expressed sequence tags analyzed, eight genes were specifically and significantly upregulated by a treatment with interferon gamma (IFN gamma) and A beta compared to a treatment with IFN gamma alone (p<0.002). We found that the gene for a well-characterized lipogenetic enzyme, stearoyl coenzyme A desaturase-1 (SCD-1), was specifically upregulated by A beta treatment and was suppressed to basal levels by RS-1178. Although the underlying mechanisms remain unknown, our results suggest the presence of a link between AD and SCD-1.