No abstract

Missense mutations in the human presenilin-1 (PS1) gene, which is found on chromosome 14, cause early-onset familial Alzheimer's disease (FAD). FAD-linked PS1 variants alter proteolytic processing of the amyloid precursor protein and cause an increase in vulnerability to apoptosis induced by various cell stresses. However, the mechanisms responsible for these phenomena are not clear. Here we report that mutations in PS1 affect the unfolded-protein response (UPR), which responds to the increased amount of unfolded proteins that accumulate in the endoplasmic reticulum (ER) under conditions that cause ER stress. PS1 mutations also lead to decreased expression of GRP78/Bip, a molecular chaperone, present in the ER, that can enable protein folding. Interestingly, GRP78 levels are reduced in the brains of Alzheimer's disease patients. The downregulation of UPR signalling by PS1 mutations is caused by disturbed function of IRE1, which is the proximal sensor of conditions in the ER lumen. Overexpression of GRP78 in neuroblastoma cells bearing PS1 mutants almost completely restores resistance to ER stress to the level of cells expressing wild-type PS1. These results show that mutations in PS1 may increase vulnerability to ER stress by altering the UPR signalling pathway.

The potential use of circularly polarized luminescence for object identification in a sensor application is demonstrated. New luminescence probes using pyrene derivatives as sensor luminophores were developed. (R,R)-Im Py and (S,S)-Im Py contain two chiral imidazole moieties at 1,6-positions through ethynyl spacers (angle between spacers ca. 180°). The probe molecules spontaneously self-assemble into chiral stacks (P or M helicity) upon coordination to metal ions with tetrahedral coordination (Zn ). The chiral probes display neither circular dichroism (CD) nor circularly polarized luminescence (CPL) without metal ions. However, (R,R)-Im Py and (S,S)-Im Py exhibit intense chiroptical activity (CD and CPL) upon self-assembly with Zn ions. (R,R)-Im Py and (S,S)-Im Py with chemical stimuli-responsibility allow sensing using the CPL signal as detection output, enabling us to discriminate between a signal from the target analyte and that from non-target species.

Mouse embryonic stem (ES) cells can proliferate indefinitely in an undifferentiated state in the presence of leukemia inhibitory factor (LIF), or differentiate into all three germ layers upon removal of this factor. To determine cellular factors associated with self-renewal of undifferentiated ES cells, we used polymerase chain reaction-assisted cDNA subtraction to screen genes that are expressed in undifferentiated ES cells and down-regulated after incubating these cells in a differentiation medium without LIF for 48 h. The mRNA expression of a tetraspanin transmembrane protein, CD9, was high in undifferentiated ES cells and decreased shortly after cell differentiation. An immunohistochemical analysis confirmed that plasma membrane-associated CD9 was expressed in undifferentiated ES cells but low in the differentiated cells. Addition of LIF to differentiating ES cells reinduced mRNA expression of CD9, and CD9 expression was accompanied with a reappearance of undifferentiated ES cells. Furthermore, activation of STAT3 induced the expression of CD9, indicating the LIF/STAT3 pathway is critical for maintaining CD9 expression. Finally, addition of anti-CD9 antibody blocked ES cell colony formation and reduced cell viability. These results indicate that CD9 may play a role in LIF-mediated maintenance of undifferentiated ES cells. INTRODUCTIONMouse embryonic stem (ES) cells, which originally derived from inner cell mass of an early embryo named blastocyst, are able to sustain their pluripotency in in vitro cell culture (Evans and Kaufman, 1981;Martin, 1981). Undifferentiated mouse ES cells can be maintained for a long time in media containing the cytokine leukemia inhibitory factor (LIF) (Smith et al., 1988;Williams et al., 1988). Pluripotency of such cultured ES cells has been demonstrated in both in vivo and in vitro experiments. When injected into blastocysts, ES cells participate in embryonic development involving all three germ layers producing chimeric mice (Bradley et al., 1984). ES cells also form teratomas containing various mature tissues when injected into immunocompromised mice (Evans and Kaufman, 1981;Martin, 1981). In vitro, mouse ES cells start to differentiate to every possible lineage upon removal of LIF from the culture medium. The mechanism by which LIF maintains ES cells in undifferentiated state is not completely understood. The transcription factor STAT3 is a downstream target of LIF and its receptor interaction Matsuda et al., 1999). It has been shown that the activity of STAT3 is necessary and sufficient for LIF-induced self-renewal of mouse ES cells (Matsuda et al., 1999). It remains unclear, however, which genes are regulated by the STAT3 transcription factor in mouse ES cells and play actual roles in the maintenance of stem cells. Indeed, there is no generally accepted mechanism by which stem cells are maintained as undifferentiated cells. Human ES cell cultures have been recently established using mouse fibroblasts as feeder cells (Shamblott et al., 1998;Thomson et al., 1998). In contrast ...