Effect of polyamine depletion on caspase activation: a study with spermine synthase-deficient cells

2001 ◽  
Vol 355 (1) ◽  
pp. 199-206 ◽  
Author(s):  
Claudio STEFANELLI ◽  
Carla PIGNATTI ◽  
Benedetta TANTINI ◽  
Monia FATTORI ◽  
Ivana STANIC ◽  
...  
Keyword(s):  
Uv Irradiation ◽  
Normal Male ◽  
Caspase Activity ◽  
Caspase Activation ◽  
Embryonic Fibroblasts ◽  
Polyamine Synthesis ◽  
Spermine Synthase ◽  
Cell Processes ◽  
Mutant Male

Activation of the caspase proteases represents a central point in apoptosis. The requirement for spermine for the processes leading to caspase activation has been studied in transformed embryonic fibroblasts obtained from gyro (Gy) mutant male mice. These cells lack spermine synthase activity and thus provide a valuable model to study the role of spermine in cell processes. Gy fibroblasts do not contain spermine and have a higher spermidine content. However, when compared with fibroblasts obtained from normal male littermates (N cells), Gy fibroblasts were observed to grow normally. The lack of spermine did not affect the expression of Bcl-2, and caspases 3 and 9 were activated by etoposide in both N and Gy cells, indicating that spermine is dispensable for caspase activation. Spermine deficiency did not significantly influence caspase activity in cells treated with etoposide, cycloheximide or staurosporine, but sensitized the cells to UV irradiation, which triggered significantly higher caspase activity in Gy cells compared with N cells. α-Difluoromethylornithine (DFMO), an inhibitor of polyamine synthesis that is able to deplete cells of putrescine and spermidine, but usually does not influence spermine content, was able to produce a more complete polyamine depletion in Gy cells. This depletion, which included spermine deficiency, dramatically increased caspase activation and cell death in Gy fibroblasts exposed to UV irradiation. On the other hand, in either N or Gy cells, DFMO treatment did not influence caspase activity triggered by staurosporine, but inhibited it when the inducers were cycloheximide or etoposide. In Gy cells depleted of polyamines by DFMO, polyamine replenishment with either spermidine or spermine was sufficient to restore caspase activity induced by etoposide, indicating that, in this model, polyamines have an interchangeable role in supporting caspase activation. Therefore, spermine is not required for such activation, and the effect and specificity of polyamine depletion on caspase activity may be very different, depending on the role of polyamines in the specific death pathways engaged by different stimuli. Some inducers of apoptosis, for example etoposide, absolutely require polyamines for caspase activation, yet the lack of polyamines, particularly spermine, strongly increases caspase activation when induced by UV irradiation.

scholarly journals Effect of spermine synthase deficiency on polyamine biosynthesis and content in mice and embryonic fibroblasts, and the sensitivity of fibroblasts to 1,3-bis-(2-chloroethyl)-N-nitrosourea

2000 ◽  
Vol 351 (2) ◽  
pp. 439-447 ◽  
Author(s):  
Caroline A. MACKINTOSH ◽  
Anthony E. PEGG
Keyword(s):  
The Body ◽  
Normal Male ◽  
Male Mice ◽  
Embryonic Fibroblasts ◽  
Primary Fibroblast ◽  
Polyamine Biosynthesis ◽  
Spermine Synthase ◽  
Fibroblast Cultures ◽  
Phex Gene ◽  
Weaning Age

Mutant Gy male mice, which have previously been described as having disruption of the phosphate-regulating Phex gene and a spermine synthase gene [Meyer, Henley, Meyer, Morgan, McDonald, Mills and Price (1998) Genomics, 48, 289–295; Lorenz, Francis, Gempel, Böddrich, Josten, Schmahl and Schmidt (1998) Hum. Mol. Genet. 7, 541–547], as well as mutant Hyp male mice, which have disruption of the Phex gene only, were examined along with their respective normal male littermates. Biochemical analyses of extracts of brains, hearts and livers of 5-week-old mice showed that Gy males lacked any significant spermine synthase activity as well as spermine content. Organs of Gy males had a higher spermidine content. This was caused not only by the lack of conversion of spermidine into spermine, but also because of compensatory increases in the activities of other polyamine biosynthetic enzymes. Gy males were half the body weight of their normal male littermates at weaning age. Hyp males, however, were no different in size when compared with their controls. High mortality of Gy males occurs by weaning age and this mortality was shown to be largely post-natal. Embryonic fibroblasts were isolated from Gy males and their normal male littermates and were similarly shown to lack any significant spermine synthase activity as well as spermine content. The lack of spermine, however, had no significant effect on the growth of immortalized fibroblasts or of primary fibroblast cultures. Similarly, there was no difference in the time of senescence of primary fibroblast cultures from Gy males compared with cultures derived from normal male littermates. However, the lack of spermine did increase the sensitivity of immortalized fibroblasts to killing by the chloroethylating agent 1,3-bis(2-chloroethyl)-N-nitrosourea. Therefore both the Gy male mice and derived embryonic fibroblasts provide valuable models to study the importance of spermine and spermine synthase, without the use of inhibitors which may have additional side effects.

scholarly journals A Functional NADPH Oxidase Prevents Caspase Involvement in the Clearance of Phagocytic Neutrophils

2007 ◽  
Vol 75 (7) ◽  
pp. 3256-3263 ◽  
Author(s):  
Rachel P. Wilkie ◽  
Margret C. M. Vissers ◽  
Mike Dragunow ◽  
Mark B. Hampton
Keyword(s):  
Nadph Oxidase ◽  
Caspase 3 ◽  
Caspase Activity ◽  
Wild Type ◽  
Caspase Activation ◽  
Effector Caspases ◽  
Nadph Oxidase Complex ◽  
Normal Human ◽  
Active Caspase

ABSTRACT Neutrophils play a prominent role in host defense. Phagocytosis of bacteria leads to the formation of an active NADPH oxidase complex that generates reactive oxygen species for bactericidal purposes. A critical step in the resolution of inflammation is the uptake of neutrophils by macrophages; however, there are conflicting reports on the mechanisms leading to the apoptosis of phagocytic neutrophils. The aim of this study was to clarify the role of effector caspases in these processes. Caspase activity was measured by DEVDase activity assays or immunofluorescence detection of active caspase-3. With normal human and wild-type murine neutrophils there was no caspase activation following phagocytosis of Staphylococcus aureus. However, caspase activity was observed in phagocytic neutrophils with a defective NADPH oxidase, including neutrophils isolated from X-linked gp91phox knockout chronic granulomatous disease mice. These results indicate that a functional NADPH oxidase and the generation of oxidants in the neutrophil phagosome prevent the activation of the cytoplasmic caspase cascade.

scholarly journals Studies of non-metabolizable polyamines that support growth of SV-3T3 cells depleted of natural polyamines by exposure to α-difluoromethylornithine

1988 ◽  
Vol 254 (2) ◽  
pp. 373-378 ◽  
Author(s):  
S Nagarajan ◽  
B Ganem ◽  
A E Pegg
Keyword(s):  
Cell Growth ◽  
Physiological Role ◽  
3T3 Cells ◽  
Polyamine Synthesis ◽  
Spermine Synthase ◽  
Support Cell ◽  
Polyamine Derivatives ◽  
Metabolic Interconversion

A number of synthetic polyamine derivatives that included five achiral gem-dimethylspermidines and two analogous tetramethylated spermines were tested for their abilities to serve as substrates for enzymes metabolizing polyamines and for their capacities to substitute for the natural polyamines in cell growth. It was found that none of the compounds were effective substrates for spermine synthase, and only one, namely 8,8-dimethylspermidine, was a substrate for spermidine/spermine N1-acetyltransferase. However, all of the spermidine derivatives and 1,1,12,12-tetramethylspermine were able to support the growth of SV-3T3 cells in which endogenous polyamine synthesis was prevented by the addition of alpha-difluoromethylornithine. These results suggest that either spermidine or spermine can support cell growth without the need for metabolic interconversion. In contrast with the result with 1,1,12,12-tetramethylspermine, 3,3,10,10-tetramethylspermine did not restore growth of polyamine-depleted SV-3T3 cells. Comparison of the properties of these derivatives may prove valuable in understanding the physiological role of polyamines.

scholarly journals Effect of spermine synthase on the sensitivity of cells to anti-tumour agents

2003 ◽  
Vol 373 (3) ◽  
pp. 885-892 ◽  
Author(s):  
Yoshihiko IKEGUCHI ◽  
Caroline A. MACKINTOSH ◽  
Diane E. McCLOSKEY ◽  
Anthony E. PEGG
Keyword(s):  
Cell Growth Rate ◽  
Mouse Embryonic Fibroblasts ◽  
Embryonic Fibroblasts ◽  
Polyamine Analogues ◽  
Spermine Synthase ◽  
Site Of Action ◽  
Cross Links ◽  
Sites Of Action ◽  
Paired Cell

The role of spermine in the sensitivity of cells to various established and experimental anti-tumour agents was examined, using paired cell lines that possess or lack spermine synthase. All spermine-synthase-deficient cells had no detectable spermine, and elevated spermidine, content. Spermine content did not alter the cell growth rate. There was little or no difference in sensitivity of immortalized mouse embryonic fibroblasts to doxorubicin, etoposide, cisplatin, methylglyoxal bis(guanylhydrazone) or H2O2 and only a slight increase in sensitivity to vinblastine and nocodazole. However, the absence of spermine clearly increased the sensitivity to 1,3-bis(2-chloroethyl)-N-nitrosourea, suggesting that depletion of spermine may be a useful way to increase the anti-neoplastic effects of anti-tumour agents that form chloroethyl-mediated interstrand DNA cross-links. The effects of spermine on the response to polyamine analogues (which have been proposed to be useful anti-neoplastic agents) were complex, and depended on the compound examined and on the cells tested. Sensitivity to CHENSpm {N1-ethyl-N11-[(cycloheptyl)methyl]-4,8-diazaundecane} was substantially greater in immortalized fibroblasts that lack spermine. In contrast, BE-3-4-3 [N1,N12-bis(ethyl)spermine] and BE-3-3-3 [N1,N11-bis(ethyl)norspermine] were more active against cells that contained spermine. The presence of spermine correlated with a greater induction of spermidine/spermine-N1-acetyltransferase by BE-3-3-3, which is consistent with suggestions that this induction is important for the response to this drug. These findings support the concepts that different polyamine analogues have different sites of action and that CHENSpm has a different site of action from BE-3-3-3.

scholarly journals Conversion of Methanol on Rutile TiO2 (110) and Tungsten Oxide Clusters: 2. The Role of Defects and Electron Transfer in Bifunctional Oxidic Photocatalysts

2021 ◽  
Author(s):  
Lars Mohrhusen ◽  
Jessica Kräuter ◽  
Katharina Al-Shamery
Keyword(s):  
Electron Transfer ◽  
Transition Metal ◽  
Metal Oxide ◽  
Tungsten Oxide ◽  
Organic Compounds ◽  
Uv Irradiation ◽  
Organic P ◽  
Rutile Tio2 ◽  
Metal Oxide Surfaces

The photochemical conversion of organic compounds on tailored transition metal oxide surfaces by (UV) irradiation has found wide applications ranging from the production of chemicals to the degradation of organic...

scholarly journals Perturbation of spermine synthase Gene Expression and Transcript Profiling Provide New Insights on the Role of the Tetraamine Spermine in Arabidopsis Defense against Pseudomonas viridiflava

2011 ◽  
Vol 156 (4) ◽  
pp. 2266-2277 ◽  
Author(s):  
María Elisa Gonzalez ◽  
Francisco Marco ◽  
Eugenio Gómez Minguet ◽  
Pedro Carrasco-Sorli ◽  
Miguel Angel Blázquez ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcript Profiling ◽  
Spermine Synthase ◽  
Pseudomonas Viridiflava ◽  
Arabidopsis Defense

scholarly journals Critical Role of PDE4D in β2-Adrenoceptor-dependent cAMP Signaling in Mouse Embryonic Fibroblasts

2008 ◽  
Vol 283 (33) ◽  
pp. 22430-22442 ◽  
Author(s):  
Matthew D. Bruss ◽  
Wito Richter ◽  
Kathleen Horner ◽  
S.-L. Catherine Jin ◽  
Marco Conti
Keyword(s):  
Critical Role ◽  
Camp Signaling ◽  
Mouse Embryonic Fibroblasts ◽  
Embryonic Fibroblasts

Selected Contribution: Differential role of nitric oxide synthase isoforms in fever of different etiologies: studies using Nosgene-deficient mice

2003 ◽  
Vol 94 (6) ◽  
pp. 2534-2544 ◽  
Author(s):  
Wieslaw Kozak ◽  
Anna Kozak
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Corn Oil ◽  
Normal Male ◽  
Wild Type ◽  
Oxide Synthase ◽  
And Control ◽  
Nos Isoforms ◽  
Deficient Mice

Male C57BL/6J mice deficient in nitric oxide synthase (NOS) genes (knockout) and control (wild-type) mice were implanted intra-abdominally with battery-operated miniature biotelemeters (model VMFH MiniMitter, Sunriver, OR) to monitor changes in body temperature. Intravenous injection of lipopolysaccharide (LPS; 50 μg/kg) was used to trigger fever in response to systemic inflammation in mice. To induce a febrile response to localized inflammation, the mice were injected subcutaneously with pure turpentine oil (30 μl/animal) into the left hindlimb. Oral administration (gavage) of N G-monomethyl-l-arginine (l-NMMA) for 3 days (80 mg · kg−1 · day−1in corn oil) before injection of pyrogens was used to inhibit all three NOSs ( N G-monomethyl-d-arginine acetate salt and corn oil were used as control). In normal male C57BL/6J mice, l-NMMA inhibited the LPS-induced fever by ∼60%, whereas it augmented fever by ∼65% in mice injected with turpentine. Challenging the respective NOS knockout mice with LPS and with l-NMMA revealed that inducible NOS and neuronal NOS isoforms are responsible for the induction of fever to LPS, whereas endothelial NOS (eNOS) is not involved. In contrast, none of the NOS isoforms appeared to trigger fever to turpentine. Inhibition of eNOS, however, exacerbates fever in mice treated with l-NMMA and turpentine, indicating that eNOS participates in the antipyretic mechanism. These data support the hypothesis that nitric oxide is a regulator of fever. Its action differs, however, depending on the pyrogen used and the NOS isoform.

Effect of UV irradiation on stabilized collagen: Role of chromium(III)

2008 ◽  
Vol 62 (1) ◽  
pp. 11-16 ◽  
Author(s):  
Nishtar Nishad Fathima ◽  
Raju Suresh ◽  
Jonnalagadda Raghava Rao ◽  
Balachandran Unni Nair ◽  
Thirumalachari Ramasami
Keyword(s):  
Uv Irradiation
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