The Experts below are selected from a list of 28881 Experts worldwide ranked by ideXlab platform
Kazuo Ohuchi - One of the best experts on this subject based on the ideXlab platform.
-
Participation of various kinases in Staurosporine induced apoptosis of RAW 264.7 cells.
Journal of Pharmacy and Pharmacology, 2002Co-Authors: Kouya Yamaki, Jangja Hong, Kenji Hiraizumi, Kazuo OhuchiAbstract:: Staurosporine induced apoptosis of RAW 264.7 cells, a mouse macrophage-like cell line, as determined by DNA fragmentation, the increase of annexin V-stained cells, and the cleavage of poly(ADP-ribose)polymerase (PARP), a substrate of caspase. Analysis of the increase in the percentage of sub-G(1) cells revealed that the DNA fragmentation occurred in a time- and concentration-dependent manner at 0.021-2.1 microM of Staurosporine. Staurosporine induced phosphorylation of p38 mitogen-activated protein kinase (MAPK) but suppressed spontaneous phosphorylation of p44/42 MAPK. The p38 MAPK inhibitor SB203580, the MAPK/extracellular signal-regulated kinase kinase inhibitor PD98059 and the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 potentiated the Staurosporine-induced PARP cleavage and DNA fragmentation. The protein kinase A (PKA) inhibitor H-89 potentiated the Staurosporine-induced DNA fragmentation without potentiating the PARP cleavage. In contrast, the protein kinase C (PKC) inhibitor Ro-31-8425 suppressed the PARP cleavage and DNA fragmentation. These findings suggested that Staurosporine induces apoptosis via the caspase cascade in RAW 264.7 cells. The Staurosporine-induced apoptosis is positively regulated by PKC, negatively regulated by p38 MAPK, p44/42 MAPK and PI3K via the caspase cascade, and negatively regulated by PKA without regulation of caspase activation.
-
Induction of neutrophil chemotactic factor production by Staurosporine in rat peritoneal neutrophils
British Journal of Pharmacology, 1997Co-Authors: Takeo Edamatsu, Yi-qun Xiao, Jun-ichi Tanabe, Kazuo OhuchiAbstract:1 Incubation of rat peritoneal neutrophils in medium containing various concentrations of Staurosporine (6.4–64 nM) increased the neutrophil chemotactic activity in the conditioned medium in a time- and concentration-dependent manner. 2 Separation of the neutrophil chemotactic activity in the conditioned medium by isoelectric focusing revealed that Staurosporine (64 nM) stimulated the production of basic (pH>8) neutrophil chemotactic factors, while TPA (12-O-tetradecanoylphorbol 13-acetate, 49 nM) stimulated the production of both basic (pH>8) and acidic (pH 5) neutrophil chemotactic factors. 3 Determination by immunoassay of cytokine-induced neutrophil chemoattractant (CINC)-1, -2α, -2β and -3 in the conditioned medium at 4 h revealed that Staurosporine (64 nM) and TPA (49 nM) strongly stimulated the production of CINC-3 (Staurosporine, 133.0±3.8; TPA, 26.7±1.0; control, 0.32±0.01 ng ml−1, means±s.e.mean from four samples) compared to CINC-1 (Staurosporine, 55.0±1.2; TPA, 12.2±0.3; control, 0.56±0.01 ng ml−1), and CINC-2α (Staurosporine, 1.09±0.03; TPA, 0.90±0.02; control,
-
Stimulation of prostaglandin E_2 production and induction of specific protein synthesis in rat peritoneal macrophages by a tumor promoter Staurosporine
Journal of Cancer Research and Clinical Oncology, 1993Co-Authors: Toshiya Tamura, Noriyasu Hirasawa, Masako Watanabe, Kazuo OhuchiAbstract:Staurosporine is a microbial anti-fungal alkaloid having potent inhibitory activity on protein kinase C and is a non 12- O -tetradecanoylphorbol-13-acetate-type tumor promoter in two-stage carcinogenesis experiments in mouse skin. Effects of Staurosporine and its structurally related compounds K-252a, KT5720 and KT5822 on prostaglandin E_2 production, release of arachidonic acid from membrane phospholipids, and uptake of [^35S]methionine into intracellular proteins were examined in rat peritoneal macrophages. Among the four compounds, only Staurosporine stimulated the production of prostaglandin E_2 and release of arachidonic acid at concentrations of 1 ng/ml and 10 ng/ml. The uptake of [^35S]methionine into cellular proteins, estimated to be 120 kDa and 125 kDa molecular mass, was also stimulated by Staurosporine treatment, and the uptake was increased in parallel with the increase in prostaglandin E_2 production. At higher concentrations (100 ng/ml and 1000 ng/ml), Staurosporine inhibited prostaglandin E_2 production and did not induce the specific protein synthesis. Other compounds neither stimulated prostaglandin E_2 production nor induced specific protein synthesis. K-252a inhibited prostaglandin E_2 production at concentrations above 10 ng/ml. These results suggest that the Staurosporine-induced proteins might participate in the tumor promotion or at least in the Staurosporine-induced stimulation of prostaglandin E_2 production.
-
Dual effects of Staurosporine on arachidonic acid metabolism in rat peritoneal macrophages
Biochimica et Biophysica Acta, 1990Co-Authors: Masako Watanabe, Susumu Tsurufuji, Toshiya Tamura, Mariko Ohashi, Noriyasu Hirasawa, Tsuneo Ozeki, Hirota Fujiki, Kazuo OhuchiAbstract:Abstract Staurosporine is a microbial anti-fungal alkaloid having a most potent inhibitory activity on protein kinase C and is recently found as a non-12-O-tetradecanoylphorbol-13-acetate (non-TPA)-type tumor promoter of mouse skin, although tumor promotion induced by a TPA-type tumor promoter teleocidin is suppressed by Staurosporine. When rat peritoneal macrophages were incubated in the medium containing various concentrations of Staurosporine, prostaglandin E2 production and release of radioactivity from [3H]arachidonic acid-labeled macrophages were stimulated at concentrations of 1 and 10 ng/ml. But higher concentrations of Staurosporine such as 100 and 1000 ng/ml showed no stimulative effect on prostaglandin E2 production although cytoplasmic free calcium levels were increased in a dose-dependent manner. Staurosporine-induced stimulation of prostaglandin E2 production was inhibited by treatment with cycloheximide, suggesting that a certain protein synthesis is prerequisite for the stimulation of arahcidonic acid metabolism. At higher concentrations (100 and 1000 ng/ml), Staurosporine inhibited TPA-type tumor promoter (TPA, teleocidin and aplysiatoxin)-induced stimulation of arachidonic acid metabolism probably due to the inhibition of protein kinases. Tumor promotion activity and anti-tumor promotion activity of Staurosporine might be explained by the fact that the lower concentrations of Staurosporine stimulate arachidonic acid metabolism and the higher concentrations of Staurosporine inhibit the tumor promoter-induced arachidonic acid metabolism, respectively.
Jochen H M Prehn - One of the best experts on this subject based on the ideXlab platform.
-
ca2 and reactive oxygen species in Staurosporine induced neuronal apoptosis
Journal of Neurochemistry, 2002Co-Authors: Jochen H M Prehn, Joaquin Jordan, Ghanashyam D Ghadge, Elke Preis, Maria F Galindo, Raymond P Roos, Josef Krieglstein, Richard J MillerAbstract:: Staurosporine (0.03-0.5 microM) induced a dose-dependent, apoptotic degeneration in cultured rat hippocampal neurons that was sensitive to 24-h pretreatments with the protein synthesis inhibitor cycloheximide (1 microM) or the cell cycle inhibitor mimosine (100 microM). To investigate the role of Ca2+ and reactive oxygen species in Staurosporine-induced neuronal apoptosis, we overexpressed calbindin D28K, a Ca2+ binding protein, and Cu/ Zn superoxide dismutase, an antioxidative enzyme, in the hippocampal neurons using adenovirus-mediated gene transfer. Infection of the cultures with the recombinant adenoviruses (100 multiplicity of infection) resulted in a stable expression of the respective proteins assessed 48 h later. Overexpression of both calbindin D28K and Cu/Zn superoxide dismutase significantly reduced Staurosporine neurotoxicity compared with control cultures infected with a beta-galactosidase overexpressing adenovirus. Staurosporine-induced neuronal apoptosis was also significantly reduced when the culture medium was supplemented with 10 or 30 mM K+, suggesting that Ca2+ influx via voltage-sensitive Ca2+ channels reduces this apoptotic cell death. In contrast, neither the glutamate receptor agonist NMDA (1-10 microM) nor the NMDA receptor antagonist dizocilpine (MK-801; 1 microM) was able to reduce Staurosporine neurotoxicity. Cultures treated with the antioxidants U-74500A (1-10 microM) and N-acetylcysteine (100 microM) also demonstrated reduced Staurosporine neurotoxicity. These results suggest a fundamental role for both Ca2+ and reactive oxygen species in staurosprine-induced neuronal apoptosis.
-
Staurosporine induced apoptosis of cultured rat hippocampal neurons involves caspase 1 like proteases as upstream initiators and increased production of superoxide as a main downstream effector
The Journal of Neuroscience, 1998Co-Authors: Aaron J Krohn, Elke Preis, Jochen H M PrehnAbstract:We induced apoptosis in cultured rat hippocampal neurons by exposure to the protein kinase inhibitor Staurosporine (30 nm, 24 hr). Treatment with the antioxidant (±)-α-tocopherol (100 μm) or the superoxide dismutase-mimetic manganese tetrakis (4-benzoyl acid) porphyrin (1 μm) significantly reduced Staurosporine-induced cell death. Using hydroethidine-based digital videomicroscopy, we observed a significant increase in intracellular superoxide production that peaked 6–8 hr into the Staurosporine exposure. This increase occurred in the absence of gross mitochondrial depolarization monitored with the voltage-sensitive probe tetramethylrhodamine ethyl ester. We then prepared extracts from Staurosporine-treated hippocampal neurons and monitored cleavage of acetyl-Tyr-Val-Ala-Asp-aminomethyl-coumarin and acetyl-Asp-Glu-Val-Asp-AMC, fluorogenic substrates for caspase-1-like and caspase-3-like proteases, respectively. Staurosporine caused a significant increase in caspase-1-like activity that preceded intracellular superoxide production and reached a maximum after 30 min. Caspase-3-like activity paralleled intracellular superoxide production, with peak activity seen after 8 hr. Treatment with the corresponding caspase-3-like protease inhibitor acetyl-Asp-Glu-Val-Asp-aldehyde (10 μm) prevented the increase in caspase-3-like activity and Staurosporine-induced nuclear fragmentation, but failed to prevent the rise in superoxide production and subsequent cell death. In contrast, treatment with caspase-1-like protease inhibitors reduced both superoxide production and cell death. Of note, antioxidants prevented superoxide production, caspase-3-like protease activity, and cell death even when added 4 hr after the onset of the Staurosporine exposure. These results suggest a scenario of an early, caspase-1-like activity followed by a delayed intracellular superoxide production that mediates Staurosporine-induced cell death of cultured rat hippocampal neurons.
Arnaldo Videira - One of the best experts on this subject based on the ideXlab platform.
-
modulation of fungal sensitivity to Staurosporine by targeting proteins identified by transcriptional profiling
Fungal Genetics and Biology, 2011Co-Authors: Andreia S Fernandes, Pedro A Goncalves, Ana Catarina Vieira De Castro, Telma Lopes, Rui Gardner, Louise N Glass, Arnaldo VideiraAbstract:An analysis of the time-dependent genetic response to the death-inducer Staurosporine was performed in Neurospora crassa by transcriptional profiling. Staurosporine induced two major genes encoding an ABC transporter and a protein with similarity to regulatory subunits of potassium channels. The transcriptional response is dependent on the activity of a novel transcription factor. Deletion mutants in differentially expressed genes displayed altered sensitivity to Staurosporine, underscoring significant proteins involved in the response to the drug. A null-mutant of the ABC transporter (abc3) is extremely sensitive to Staurosporine, accumulates more Staurosporine than the wild type strain and is defective in energy-dependent export of the drug, indicating that the ABC3 protein is the first described Staurosporine transporter. It was located in the plasma membrane by immunofluorescence microscopy. The combination of inhibitors of ABC transporters or of potassium channels with Staurosporine leads to an enhanced activity against N. crassa and pathogenic fungi paving the way to the development of more potent and specific antifungals. Our results highlight the general use of transcriptional profiling for the identification of novel proteins involved in cell death and their potential use as drug targets.
-
rotenone enhances the antifungal properties of Staurosporine
Eukaryotic Cell, 2010Co-Authors: Andreia S Fernandes, Ana Catarina Vieira De Castro, Arnaldo Videira, Catarina Lemos, A Falcao, N L GlassAbstract:We studied Staurosporine-induced cell death in the filamentous fungus Neurospora crassa. The generation of reactive oxygen species during the process appears to be an important signaling event, since addition of the antioxidant glutathione prevents the effects of Staurosporine on fungal growth. Selected mutants with mutations in respiratory chain complex I are extremely sensitive to the drug, stressing the involvement of complex I in programmed cell death. Following this finding, we determined that the complex I-specific inhibitor rotenone used in combination with Staurosporine results in a synergistic and specific antifungal activity, likely through a concerted action on intracellular glutathione depletion. Paradoxically, the synergistic antifungal activity of rotenone and Staurosporine is observed in N. crassa complex I mutants and in Saccharomyces cerevisiae, which lacks complex I. In addition, it is not observed when other complex I inhibitors are used instead of rotenone. These results indicate that the rotenone effect is independent of complex I inhibition. The combination of rotenone and Staurosporine is effective against N. crassa as well as against the common pathogens Aspergillus fumigatus and Candida albicans, pointing to its usefulness as an antifungal agent.
Richard J Miller - One of the best experts on this subject based on the ideXlab platform.
-
ca2 and reactive oxygen species in Staurosporine induced neuronal apoptosis
Journal of Neurochemistry, 2002Co-Authors: Jochen H M Prehn, Joaquin Jordan, Ghanashyam D Ghadge, Elke Preis, Maria F Galindo, Raymond P Roos, Josef Krieglstein, Richard J MillerAbstract:: Staurosporine (0.03-0.5 microM) induced a dose-dependent, apoptotic degeneration in cultured rat hippocampal neurons that was sensitive to 24-h pretreatments with the protein synthesis inhibitor cycloheximide (1 microM) or the cell cycle inhibitor mimosine (100 microM). To investigate the role of Ca2+ and reactive oxygen species in Staurosporine-induced neuronal apoptosis, we overexpressed calbindin D28K, a Ca2+ binding protein, and Cu/ Zn superoxide dismutase, an antioxidative enzyme, in the hippocampal neurons using adenovirus-mediated gene transfer. Infection of the cultures with the recombinant adenoviruses (100 multiplicity of infection) resulted in a stable expression of the respective proteins assessed 48 h later. Overexpression of both calbindin D28K and Cu/Zn superoxide dismutase significantly reduced Staurosporine neurotoxicity compared with control cultures infected with a beta-galactosidase overexpressing adenovirus. Staurosporine-induced neuronal apoptosis was also significantly reduced when the culture medium was supplemented with 10 or 30 mM K+, suggesting that Ca2+ influx via voltage-sensitive Ca2+ channels reduces this apoptotic cell death. In contrast, neither the glutamate receptor agonist NMDA (1-10 microM) nor the NMDA receptor antagonist dizocilpine (MK-801; 1 microM) was able to reduce Staurosporine neurotoxicity. Cultures treated with the antioxidants U-74500A (1-10 microM) and N-acetylcysteine (100 microM) also demonstrated reduced Staurosporine neurotoxicity. These results suggest a fundamental role for both Ca2+ and reactive oxygen species in staurosprine-induced neuronal apoptosis.
Mark P Mattson - One of the best experts on this subject based on the ideXlab platform.
-
calcium and reactive oxygen species mediate Staurosporine induced mitochondrial dysfunction and apoptosis in pc12 cells
Journal of Neuroscience Research, 1998Co-Authors: Inna I Kruman, Mark P MattsonAbstract:The bacterial alkaloid Staurosporine is widely employed as an inducer of apoptosis in many cell types including neurons. The intracellular cascades that mediate Staurosporine-induced apoptosis are largely unknown. Exposure of cultured PC12 cells to Staurosporine resulted in a rapid (min) and prolonged (1–6 hr) elevation of intracellular free calcium levels [Ca2+]i, accumulation of mitochondrial reactive oxygen species (ROS), and decreased mitochondrial 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction (1–4 hr). These early events were followed by membrane lipid peroxidation, loss of mitochondrial transmembrane potential, and nuclear apoptotic changes. Treatment of cells with serum or nerve growth factor within 1–2 hr of Staurosporine exposure resulted in recovery of [Ca2+]i and ROS levels, and rescued the cells from apoptosis. The increased [Ca2+]i and ROS production were required for Staurosporine-induced apoptosis because the intracellular calcium chelator BAPTA and uric acid (an agent that scavenges peroxynitrite) each protected cells against apoptosis. The caspase inhibitor zVAD-fmk and the anti-apoptotic gene product Bcl-2 prevented the sustained [Ca2+]i increase and ROS accumulation induced by Staurosporine indicating that caspases act very early in the apoptotic process. Our data indicate that a [Ca2+]i increase is an early and critical event in Staurosporine-induced apoptosis that engages a cell death pathway involving ROS production, oxidative stress, and mitochondrial dysfunction. J. Neurosci. Res. 51:293–308, 1998. © 1998 Wiley-Liss, Inc.
