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Y Tsujimoto

Publications and source records attributed to Y Tsujimoto.

At least 109 records · Page 6Linked to original sources
PubMed1996

Localization of bcl-2, bax, and bcl-x mRNAs in the developing inner ear of the mouse.

In situ hybridization was employed to study the expression of bcl-2 mRNA and its family members, bax and bcl-x mRNAs, in the developing inner ear. We found that in the cochlear structure, sensory epithelial cells, the spiral ganglion and stria vascularis expressed these mRNAs in postnatal period in a temporally similar manner, but in embryos, neither bax nor bcl-x mRNA were expressed in the sensory epithelium from embryonic day (E) 13 to 19. In contrast to these patterns, bcl-2 mRNA was expressed by E15 to E19, and the expression at E13 was below the lower limit of detection. Non-neuronal tissue (stria vascularis) also expressed these three transcripts during development. These results suggest that bcl-2 family members may be differentially involved in the differentiation of sensory epithelial cells, spiral ganglia, and stria vascularis. In particular, the differential expression patterns in the cochleovestibular neurons suggest that proliferating and differentiating neurons utilize distinct members of the bcl-2 family.

Animals↗
PubMed1996

Bcl-2 blocks loss of mitochondrial membrane potential while ICE inhibitors act at a different step during inhibition of death induced by respiratory chain inhibitors.

Bcl-2, Bcl-xL, CrmA and tetrapeptide ICE inhibitor reduce the extent of necrotic cell death induced by cyanide, which primarily damages mitochondria. Although none of them affects the drastic decrease in ATP levels induced by cyanide, Bcl-2 and Bcl-xL but not CrmA or ICE inhibitor inhibit the cyanide-induced decrease in mitochondrial membrane potential. A similar blocking effect is observed on necrotic cell death induced by other respiration inhibitors, rotenone and antimycin A, and on apoptotic cell death induced by etoposide or calcium ionophore. These results indicate that Bc1-2 and Bcl-xL protect mitochondria against the loss of function during both apoptosis and at least some forms of necrotic cell death. The ICE family proteases act at a different step other than the loss of mitochondrial membrane potential.

Adenosine Triphosphate↗
PubMed1996

Bcl-2 expression prevents activation of the ICE protease cascade.

The Bcl-2 family and the ICE family of cysteine proteases play important roles in regulating cell death. We show here that induction of cell death by a Ca2+ ionophore or hypoxia results in increased levels and activity of active ICE(-like) proteases and the subsequent activation of CPP32/Yama(-like) proteases, and that inhibition of these protease activities reduces the extent of cell death. Overexpression of the anti-apoptotic proteins Bcl-2 or Bcl-xL inhibits the cell death and the activation of ICE(-like) and CPP32/Yama(-like) proteases, indicating that Bcl-2 and Bcl-xL act upstream of these proteases. We also show that specific inhibition of ICE(-like) proteases in vivo prevents activation of CPP32/Yama(-like) proteases, whereas inhibition of CPP32/Yama(-like) proteases does not prevent activation of ICE(-like) proteases, suggesting the existence of a protease cascade in vivo that requires ICE(-like) proteases for activation of CPP32/Yama(-like) proteases. Induction of necrotic cell death by KCN also induces activation of ICE(-like) proteases but not of CPP32/Yama(-like) proteases, and Bcl-2 and Bcl-xL inhibit the activation and the cell death, suggesting that the functional site of Bcl-2 and Bcl-xL is also upstream of ICE(-like) proteases in at least some forms of necrosis.

Adrenal Gland Neoplasms↗
PubMed1996

Retardation of chemical hypoxia-induced necrotic cell death by Bcl-2 and ICE inhibitors: possible involvement of common mediators in apoptotic and necrotic signal transductions.

Inhibition of the respiratory chain reaction by cyanide, rotenone or antimycin A (chemical hypoxia) induces necrotic cell death characterized by apparently intact chromatin, remarkable mitochondrial swelling with loss of crista structure, and loss of plasma membrane integrity. The treatments induce no apoptotic cell death, as defined by fragmented nuclei with condensed chromatin, fragmented or condensed cytoplasm. The anti-apoptotic proteins Bcl-2 and Bcl-xL effectively retard the chemical hypoxia-induced necrotic cell death. The necrotic cell death is also retarded by inhibitors of ICE(-like) proteases, including interleukin-1beta converting enzyme (ICE), which are common mediators of apoptosis. These results indicate that Bcl-2/Bcl-xL and ICE(-like) proteases modulate apoptotic and at least some forms of necrotic cell death. Both cell death pathways appear to involve some common mediators; however necrotic or apoptotic cell death signals might be transduced through multiple pathways, because Bcl-2/ Bcl-xL or inhibitors of ICE(-like) proteases are relatively less potent in blocking necrotic cell death than in preventing apoptosis.

Amino Acid Sequence↗
PubMed1996

Induction of apoptosis as well as necrosis by hypoxia and predominant prevention of apoptosis by Bcl-2 and Bcl-XL.

The molecular mechanism of cell death due to hypoxia has not been elucidated. Our recent observations that overexpression of the anti-apoptotic proto-oncogene bcl-2 and a bcl-2-related gene, bcl-x, prevents hypoxic cell death suggest that hypoxia induces apoptosis. Using electron microscopy and confocal and nonconfocal fluorescence microscopy, we show here that hypoxia does, in fact, induce both necrosis and apoptosis, and that the proportion of these two modes is highly dependent on the cell type. Overexpression of Bcl-2 or Bcl-Xl blocks hypoxia-induced apoptosis in a dose-dependent manner.

Animals↗
PubMed1996

Involvement of CPP32/Yama(-like) proteases in Fas-mediated apoptosis.

Fas (Apo-1/CD95) belongs to the tumor necrosis factor/nerve growth factor receptor family and transmits apoptotic signals by binding to its ligand. Interleukin-1beta-converting enzyme (ICE), which shows substantial homology to the product of the cell death gene, ced-3, of Caenorhabditis elegans, is reported to be involved in Fas-mediated apoptosis. Using two human carcinoma-derived cell lines with undetectable levels of ICE, we found that an agonistic antihuman Fas antibody induces the activation of CPP32/Yama(-like) proteases that are ICE(-like) protease family members, and that a tetrapeptide inhibitor of CPP32/Yama protease, DEVD-CHO, inhibits the Fas-mediated activation of the proteases, Fas-mediated apoptosis, and CPP32/Yama(-like) proteolytic activities in vitro. Fas-mediated apoptosis is inhibited by the CPP32/Yama inhibitor DEVD-CHO, but not by the ICE inhibitor YVAD-CHO, suggesting a dominant role for the CPP32/Yama(-like) proteases and not ICE itself in Fas-mediated apoptosis of the human carcinoma cell lines.

Amino Acid Sequence↗
PubMed1996

Involvement of Fas in regression of vaginal epithelia after ovariectomy and during an estrous cycle.

Fas, also called APO-1, belongs to the tumor necrosis factor/nerve growth factor receptor family and transmits an apoptotic signal within the cell by binding to the Fas ligand. Fas has been implicated in the activation-induced suicide of T cells and cytotoxic T cell activity in the immune system. Non-immune cells such as those in liver, lung and ovary also express Fas, but its role in these cells remains unclear. Ovariectomy has been used to study homeostasis of female reproductive organs, which is regulated by sex hormones. Here we analyzed Fas function in the ovariectomy-induced regression of mouse vaginal epithelial cells. Fas expression was detected in vagina and was elevated after ovariectomy. Fas-deficient lpr and lpr(cg) mice did not exhibit ovariectomy-induced regression of vaginal epithelia, whereas uterine regression induced by ovariectomy was not affected in these mice. The vaginas of lpr and lpr(cg) mice were in a persistent estrous stage with cornification of vaginal epithelia, as judged from the cell types in the vaginal fluid. Thus, Fas appears to be involved directly in the regression of vaginal epithelia induced by ovariectomy and during the estrous cycle, suggesting that the physiological role of this receptor extends beyond that exerted on immune cells. This is the first evidence of a role for Fas inducing physiological apoptosis in non-immune cells.

Animals↗
PubMed1996

bcl-2: antidote for cell death.

The bcl-2 proto-oncogene, originally identified through the study of the t(14;18) translocation present in human B-cell follicular lymphomas, is unique among oncogenes in its ability to enhance cell survival by interfering with apoptotic cell death. This finding provided the important notion to the cancer research field that inhibition of cell death might be a critical step in tumorigenesis. This idea was supported by the demonstration that several cancer genes, including oncogenes and anti-oncogenes, have activities to modulate the apoptotic process. bcl-2 exerts a death-sparing activity against apoptosis induced by a wide variety of stimuli and, therefore, appears to function at a critical step in a common process in which several different apoptotic signals converge, although the mechanism of bcl-2 function remains unknown. bcl-2 has recently been recognized as a member of a family through the discovery of many structurally related genes, some of which function like bcl-2 while others inhibit the death-sparing function of bcl-2 or other members. Detailed analysis of the bcl-2 family members will provide important clues toward an understanding of the molecular basis of apoptotic cell death. Here, current information on the bcl-2 gene and other members of this family is reviewed.

Animals↗
PubMed1996

Cytokine-induced apoptotic cell death in a mouse pancreatic beta-cell line: inhibition by Bcl-2.

Cytokines are thought to contribute to the induction of pancreatic beta-cell destruction in insulin-dependent diabetes mellitus. The molecular mechanisms that underlie beta-cell death were investigated by studying cytokine-induced cell death in beta-cell lines. A combination of three cytokines (interleukin-1 beta, tumour necrosis factor-alpha, and interferon-gamma) induced apoptotic cell death in the mouse pancreatic beta-cell line beta TC1, as judged from the appearance of cells with hypodiploid nuclei and oligonucleosomal DNA fragmentation. The same treatment also induced apoptosis in the mouse pancreatic alpha-cell line alpha TC1 and the NOD/Lt mouse beta-cell line NIT-1, although to a lesser extent than in beta TC1 cells. The abundance of endogenous Bcl-2 in beta TC1 cells was lower than that in the other two cell lines. Overexpression of human Bcl-2 in beta TC1 cells partially protected them from cytokine-induced cell death. These results suggest that apoptosis may be responsible, at least in part, for cytokine-induced beta-cell destruction and that Bcl-2 prevents apoptosis in pancreatic islet cells.

Animals↗
PubMed1996

Molecular cloning of the breakpoint of t(11;22) (q23;q11) chromosome translocation in an adult acute myelomonocytic leukaemia.

Southern blot analysis with a cDNA probe of MLL indicated that the breakpoint is in a BamHI 8.3 kb fragment which carries the exon 5-11 of MLL gene in DNA from an adult acute myelomonocytic leukaemia with a t(11;22) (q23;q11) translocation. The structural analysis of the rearranged MLL locus demonstrated that the breakpoint is localized between exon 8 and 9 of MLL locus. The normal counterpart fused to the MLL locus was proved to be derived from chromosome 22q11(AF-22) by somatic cell hybrids analysis and FISH. By FISH, AF-22 was localized to the region more centromeric to the BCR gene.

Adult↗
PubMed1996

Involvement of ICE family proteases in apoptosis induced by reoxygenation of hypoxic hepatocytes.

Cell death due to reoxygenation after hypoxia was characterized in primary cultured hepatocytes. Fluorescence and electron microscopic analyses of reoxygenated hepatocytes revealed morphological characteristics of apoptosis, including chromatin condensation, nuclear fragmentation, and formation of apoptotic bodies. Few necrotic hepatocytes, defined by loss of plasma membrane integrity, mitochondrial swelling, and formation of large vacuoles, were observed. Activation of interleukin-1 beta-converting enzyme (ICE)-like and CPP32/Yama-like proteases, which are known to drive apoptosis, was observed during reoxygenation, and addition of their respective inhibitors inhibited the induction of apoptosis, indicating the involvement of ICE family proteases in apoptosis by reoxygenation. Production of oxygen radicals was enhanced by reoxygenation of hypoxic cells, and reoxygenation-induced apoptosis was inhibited by oxygen radical scavengers, suggesting a role for reactive oxygen species as a triggering factor in cell death. Electrophoretic analysis revealed the presence of 50-kb DNA fragments but not oligonucleosomal DNA fragments in reoxygenation-induced apoptotic hepatocytes.

Animals↗
PubMed1996

Chromosomal assignment of the Bcl2-related genes, Bcl2l and Bax, in the mouse and rat.

The Bcl2-related genes, Bax and Bcl2l (formerly called Bclx), are involved in the regulation of apoptotic cell death, and the chromosome location of these two genes was determined in the mouse and rat by fluorescence in situ hybridization. Bcl2l was localized to mouse chromosome 2H1 and rat chromosome 3q41.2, and Bax was localized to mouse chromosome 7B5 and rat chromosome 1q31.2. Both genes mapped in a region of conserved linkage homology between the two the species. Molecular linkage analysis using interspecific backcross mice revealed the murine Bcl2l locus at 0.7 cM terminal to D2Mit22, and the murine Bax locus at 0.8 cM terminal to D7Nds5.

Animals↗
PubMed1996

Nitric oxide induces upregulation of Fas and apoptosis in vascular smooth muscle.

Interleukin-1 induced a time-dependent release of high levels of nitric oxide from rat vascular smooth muscle cells up to 96 hours. A time-dependent release of lactate dehydrogenase was also induced by Interleukin-1 from 72 to 96 hours after its stimulation. In situ nick end-labeling assay revealed that incubation for 48 hours with interleukin-1 induced a positive staining of fragmented nuclei. However, NG-monomethyl-L-arginine, an inhibitor of nitric oxide synthase, inhibited both lactate dehydrogenase release and DNA fragmentation induced by interleukin-1. Furthermore, sodium nitroprusside, a nitric oxide donor, also induced lactate dehydrogenase release and DNA fragmentation. Fluorescent staining of DNA revealed patches of irregularly dispersed, brightly staining, and condensed chromatin in rat vascular smooth muscle cells treated with sodium nitroprusside. Flow cytometric analysis with monoclonal antibody against human Fas revealed that expression of Fas was upregulated by sodium nitroprusside in human vascular smooth muscle cells. Methylene blue, an inhibitor of soluble guanylate cyclase, did not affect sodium nitroprusside-induced upregulation of Fas. Furthermore, 8-bromo-guanosine 3':5'-cyclic monophosphate, an analogue of cGMP, did not upregulate Fas expression. These findings indicate that nitric oxide released from vascular smooth muscle cells may induce apoptosis in vascular smooth muscle cells themselves and also induced upregulation of Fas via a cGMP-independent mechanism. Thus, nitric oxide could trigger the remodeling of atherosclerotic plaques.

Animals↗
PubMed1996

Oxidative stress response in yeast: purification and characterization of glutathione reductase from Hansenula mrakii.

Glutathione reductase was purified from a yeast. Hansenula mrakii IFO 0895, to approximately 3500-fold with 59% activity yield. The enzyme was homogeneous on polyacrylamide gel electrophoresis. The molecular weight of the enzyme was estimated to be 56 kDa by SDS-polyacrylamide gel electrophoresis, and 123 kDa by gel filtration using a calibrated Sephadex G-150 column. The Km values for glutathione disulfide and NADPH were 21.3 microM and 14.3 microM, respectively. The enzyme was most active at pH 7.5, 55 degrees C. The enzyme was stable up to 40 degrees C, and between pHs 4 and 10. The enzyme was inhibited by p-chloromercuribenzoate and metal ions such as Fe3+, Cd2+, Cu2+, and Zn2+.

Chloromercuribenzoates↗
PubMed1996

[Prevention of cell death by Bcl-2 family genes].

Apoptosis is a tightly regulated and evolutionally conserved suicide mechanism. Proteases encoded by ced-3 and ice gene family appear to play a key role in driving apoptosis. On the other hand, Bcl-2 and its relatives have the ability to inhibit apoptosis induced by a variety of stimuli, indicating that Bcl-2 family negatively regulates a key event in a common pathway of apoptosis in which different apoptotic signals finally converge, although the biochemical basis of the function remains unknown. Here, I overview Bcl-2 gene family, and discuss the molecular basis of anti-apoptotic activity of Bcl-2, focusing on our recent findings on Bcl-2 function in mitochondria.

Animals↗
PubMed1996

[Inhibition of apoptosis by a baculovirus p35 gene].

The baculovirus gene p35 inhibits virus-induced apoptosis in insect cells. p35 can also inhibit developmentally programmed cell death in Caenorhabditis elegans and Drosophila, mammalian neuronal cell death induced by serum or NGF deprivation, and Fas- and tumor necrosis factor (TNF)-induced apoptosis in mammalian cells, indicating that p35 may interrupt an evolutionally conserved component of the death machinery. Recently it has been shown that p35 protein functions as an inhibitor of ICE/CED-3 cysteine protease family that seem to play an important role in an apoptotic pathway. This observation indicates that p35 may inhibit apoptosis by directly blocking the activities of these cysteine proteases in diverse animals.

Amino Acid Sequence↗
PubMed1995

Oxidative stress response in yeast: glutathione peroxidase of Hansenula mrakii is bound to the membrane of both mitochondria and cytoplasm.

The yeast Hansenula mrakii IFO 0895 induces glutathione peroxidase (GPx) when the cells are exposed to the oxidative stress such as lipid hydroperoxide, superoxide- and hydroxy radical-generating conditions. To clarify the localization of GPx in H. mrakii cell, distribution of the enzyme was investigated. After centrifugation of the yeast protoplast homogenates at 2500 x g for 10 min, 67% of total GPx activity was recovered from the supernatant (Sup. 1) and 33% was from the pellet (Pellet 1). When the Sup. 1 was fractionated by sucrose density gradient ultracentrifugation, GPx activity was essentially recovered from the mitochondria fraction. Submitochondrial localization of the enzyme showed that 95% and 2.5% of the enzyme was recovered from the inner and outer membrane, respectively. No GPx activity was detected neither in intermembrane space nor in matrix of mitochondria. On the other hand, at least 12% of total GPx activity was recovered from the purified plasma membrane which was obtained from the Pellet 1 by successive sucrose density gradient centrifugation. Thus, the GPx of H. mrakii is present in the inner and outer membrane of mitochondria as well as the plasma membrane.

Cell Membrane↗