HUGO ID Detailed Result 19986

HUGO ID 19986
Symbol CYCS
Name cytochrome c, somatic
#Occurrence 131
#Paper 20

 

PMID Match String Actual String Score Flanking text Edited by Edit
8899665cytochrome ccytochrome c1.0cytochrome c group|free radicals|hemostatics|reactive oxygen species|thiobarbituric acid reactive substances|vitamin k|superoxide dismutase|  
10593879cytochrome ccytochrome c1.0acsf was first pumped through the system at 15 microl/min for 1 h after placement of the perfusing loop then cytochrome c 50 microm in acsf was administered through the loop.  
10593879cytochrome ccytochrome c1.0the perfusates were centrifuged and reduced cytochrome c in the supernatant was measured by a spectrophotometer at a wavelength of 550 nm.  
10593879cytochrome ccytochrome c1.0to determine the levels of o 2 in the spinal cord tissue 500 microm cytochrome c was infused into the intrathecal space by the loop through the holes made in the wall at a flow rate of 1 microl/min for 30 min.  
10593879cytochrome ccytochrome c1.0the tissue was dropped into a vial containing 500 microm cytochrome c in acsf then homogenized and centrifuged; the supernatant was passed through a 30 000 molecular weight ultrafiltration membrane.  
10593879cytochrome ccytochrome c1.0a the absorbance of reduced cytochrome c measured in the perfusates collected from msod1 n =6 sod1 n =3 and nc mice n =6 .  
10593879cytochrome ccytochrome c1.0b the absorbance of reduced cytochrome c measured in spinal cord tissue in msod1 n =3 sod1 n =3 and nc mice n =4 .  
10593879cytochrome ccytochrome c1.0the levels of o 2 in the terminal cistern were sampled by a microcannula and determined by measuring reduced cytochrome c in the perfusates using our unique method 40 .  
10593879cytochrome ccytochrome c1.0after the initial acsf perfusion the perfusing fluid was changed to cytochrome c 50 microm in acsf solution and equilibrated for 30 min.  
10593879cytochrome ccytochrome c1.0reduced cytochrome c in the supernatant was measured by a spectrophotometer at a wavelength of 550 nm.  
10593879cytochrome ccytochrome c1.0to determine the level of o 2 in the spinal cord tissue 500 microm cytochrome c was infused into the intrathecal space by the loop through the holes made on the wall at a flow rate of 1 microl/min for 30 min.  
10593879cytochrome ccytochrome c1.0the tissue was removed and transferred into a vial containing 500 microm cytochrome c in acsf homogenized centrifuged at 13 000 x g for 15 min and the supernatant was passed through a 30 000 molecular weight ultrafiltration membrane micron separation inc. westborough mass. .  
10593879cytochrome ccytochrome c1.0the average absorbance of reduced cytochrome c mean_amp_#177; sd is 0.060 _amp_#177; 0.011 in msod1 mice n =6 0.037 _amp_#177; 0.004 in sod1 mice n =3 and 0.130 _amp_#177; 0.030 in nc mice n =6 .  
10593879cytochrome ccytochrome c1.0the average absorbance of reduced cytochrome c mean_amp_#177; sd absorbance/g wet weight tissue is 158 _amp_#177; 10 in msod1 mice n =3 138 _amp_#177; 10 in sod1 mice n =3 and 186 _amp_#177; 12 in nc mice n =4 .  
10671549cytochrome ccytochrome c1.0cytochrome c release from mitochondria and caspase 3 activation also occurred.  
10671549cytochrome ccytochrome c1.0among these the protein product of bcl 2 protooncogene plays a pivotal role in apoptosis by regulating cytochrome c efflux from mitochondria which in turn activates cysteine proteases caspases ultimately leading to specific dna fragmentation.  
10671549cytochrome ccytochrome c1.0the results obtained showed that gsno mediated apoptosis in these cells is associated with a canonical sequence of events including bcl 2 p53 and p21 modulation cytochrome c release in the cytosol and caspase 3 activation.  
10671549cytochrome ccytochrome c1.0obenzenesulfonamide phenylmethylsulfonyl fluoride n 1 naphthyl ethylenediamine dihydrochloride bathocuproinedisulfonic acid monoclonal anti bcl 2 clone bcl 2 100 monoclonal anti p53 clone bp53 12 and cytochrome c horse heart were obtained from sigma.  
10671549cytochrome ccytochrome c1.0anti cytochrome c monoclonal antibody clone 7h8.2c12 was from pharmingen san diego ca .  
10671549cytochrome ccytochrome c1.0cytosolic cytochrome c determination cells detached and attached were washed with pbs and collected by centrifugation at 700 _amp_#215; g for 5 min at 4 degreec.  
10671549cytochrome ccytochrome c1.0purified cytochrome c from horse heart was used as standard.  
10671549cytochrome ccytochrome c1.0purified mouse anti cytochrome c monoclonal antibody was used as primary antibody 1:5000 .  
10671549cytochrome ccytochrome c1.0the activity was measured spectrophotometrically monitoring the oxidation of cytochrome c horse heart which had previously been reduced by treatment with excess ascorbate followed by passage on sephadex g 25 resin amersham pharmacia biotech .  
10671549cytochrome ccytochrome c1.0activity was expressed as micromoles of cytochrome c oxidized per min mg protein using an extinction coefficient of 27.6 m m cm .  
10671549cytochrome ccytochrome c1.0nuclear apoptosis is preceded by the disruption of the mitochondrial transmembrane potential which in turn can facilitate the release of pro apoptotic proteins such as cytochrome c through the opening of the mitochondrial permeability transition pore 54 .  
10671549cytochrome ccytochrome c1.0in a previous report regarding another cell model system undergoing apoptosis 56 we demonstrated that cytochrome c release from mitochondria followed oxidative stress consequent to glutathione depletion.  
10671549cytochrome ccytochrome c1.0furthermore this phenomenon could be observed in healthy cells as well suggesting that cytochrome c release is part of a more general mechanism related to redox unbalance.  
10671549cytochrome ccytochrome c1.0in contrast to this hypothesis in the present report gsno treatment led to cytochrome c release with the maintenance of high intracellular glutathione concentration.  
10671549cytochrome ccytochrome c1.0moreover in the g93a cells a small level of released cytochrome c was also detected in the untreated cells indicating that these cells in culture may spontaneously die by apoptosis.  
10671549cytochrome ccytochrome c1.0taken together these results point to a mechanism for cytochrome c release different from that related to glutathione depletion.  
10671549cytochrome ccytochrome c1.0a cytochrome c released in the cytosol was detected by western blotting using a monoclonal antibody as described under "experimental procedures." 50 microg of cytosolic protein were loaded on each lane.  
10671549cytochrome ccytochrome c1.0lane 1 sh sy5y cells; lane 2 gsno treated; lane 3 g93a cells; lane 4 gsno treated; lane 5 wt cells; lane 6 gsno treated; lane 7 purified cytochrome c.  
10671549cytochrome ccytochrome c1.0apoptotic markers: cytochrome c release caspase activation p53 accumulation p21 increase and bcl2 down regulation to examine the sequence of events occurring upon gsno toxicity we measured some of the molecular markers of apoptotic 
10671549cytochrome ccytochrome c1.0cytosolic extracts were prepared under conditions that keep mitochondria intact and cytosolic cytochrome c protein levels were measured by immunoblot analysis.  
10671549cytochrome ccytochrome c1.0the cytosol from untreated sh sy5y and wt cells contained no detectable amounts of cytochrome c whereas untreated g93a cells had a small level of released cytochrome c probably as result of spontaneous apoptotic death fig 3 a .  
10671549cytochrome ccytochrome c1.0cytosolic cytochrome c accumulated after gsno treatment; the release was highest for g93a cells and almost at the limit of detection for wt cells.  
10671549cytochrome ccytochrome c1.0caspase 3 activation followed the same trend as cytochrome c release with higher activity in the cytosolic extracts of g93a cells than in sh sy5y and wt cells fig 3 b .  
10671549cytochrome ccytochrome c1.0cytochrome c group|nitroso compounds|proto oncogene proteins c bcl 2|tumor suppressor protein p53|nitric oxide|s nitrosoglutathione|glutathione|superoxide dismutase|caspases|oncogene protein p2|  
11050436cytochrome ccytochrome c1.0complex iii ubiquinol cytochrome c oxidoreductase is responsible for taking reducing equivalents which are generated in complexes i and ii and contained in ubiquinol and transfer ring them through reactions with cytochrome b the riesk 
11050436cytochrome ccytochrome c1.0ble for taking reducing equivalents which are generated in complexes i and ii and contained in ubiquinol and transfer ring them through reactions with cytochrome b the rieske iron sulphur protein and cytochrome c 1 to the final electron acceptor cytochrome c .  
11050436cytochrome ccytochrome c1.0 1 to the final electron acceptor cytochrome c .  
11679167cytochrome ccytochrome c1.0in agreement s. cerevisiae mutants deficient in antioxidant defences such as catalase superoxide dismutase and cytochrome c peroxidase are sensitive to a lethal heat shock and the overexpression of genes encoding catalase and superoxide dismutase increase the resistance to the severe heat shock davidson et al. 1996 .  
11679167cytochrome ccytochrome c1.0the activation of these factors requires a functional mitochondria and the activity of cytochrome c peroxidase ccp1p .  
11679167cytochrome ccytochrome c1.0it has been suggested that the cytochrome c released into the cytosol as a consequence of pore opening has a pro apoptotic activity manon et al. 1997 ; minn et al. 1999 .  
11679167cytochrome ccytochrome c1.0however ant is not essential for bax induced cytochrome c release from the mitochondria and contradictory results have been described regarding the role of vdac priault and priault ; shimizu et al. 2000 .  
11679167cytochrome ccytochrome c1.0it should be noted that the importance of cytochrome c release in bax lethality was recently questioned as bax induces cell death in yeast cells expressing a functional cytochrome c_amp_#x2013;gfp fusion that is not relocalised into the cytosol roucou et 
11905995cytochrome ccytochrome c1.0ll culture system we demonstrated that infection of mouse nsc 34 motor neuron like cells with adenovirus containing mutant g93a sod1 gene increased cellular oxidative stress mitochondrial dysfunction cytochrome c release and motor neuron cell death.  
12368231cytochrome ccytochrome c1.0overexpression of sod1 protects vulnerable motor neurons after spinal cord injury by attenuating mitochondrial cytochrome c release.  
12368231cytochrome ccytochrome c1.0superoxide production mitochondrial release of cytochrome c and activation of caspase 9 were examined and apoptotic dna injury was also characterized.  
12368231cytochrome ccytochrome c1.0in the wild type animals increased superoxide production mitochondrial release of cytochrome c and cleaved caspase 9 were observed exclusively in vmn after sci.  
12368231cytochrome ccytochrome c1.0transgenic animals showed less superoxide production mitochondrial cytochrome c release and caspase 9 activation resulting in death of only 45% of the vmn.  
12368231cytochrome ccytochrome c1.0cytochrome c group|superoxides|superoxide dismutase 1|superoxide dismutase|  
14698606cytochrome ccytochrome c1.0they are also compatible with possible apoptotic contributions to motor neuron loss in als [ 58 ] because mitochondria which contain cytochrome c and other apoptotic mediators are crucial regulators of apoptosis.  
14739060cytochrome ccytochrome c1.0_amp_#x2022; cytochrome c a member of the mitochondrial electron chain required for the generation of atp when it is released from the mitochondria into cytoplasm trigger the caspase chain _amp_#x2022; smac/diablo binds to in 
14739060cytochrome ccytochrome c1.0this results in the collapse of trans membrane electrochemical gradient the loss of matrix solutes the swelling of mitochondria causing the release of cytochrome c procapases 2 3 and 9 apoptosis initiating factor and caspase activated dnase.  
14739060cytochrome ccytochrome c1.0cytochrome c and the cytosolic factor apaf1 activate the caspases while apoptosis initiating factor and caspase activated dnase move to the nucleus initiating apoptosis or programmed cell death [ 27 ].  
15896810cytochrome ccytochrome c1.0other heme protein targets for no are catalase cytochrome c hemoglobin and peroxidase.  
15964487cytochrome ccytochrome c1.0opening of a permeability transition pore ptp leads to mitochondrial swelling and the release of intramitochondrial proteins to the cytoplasm including cytochrome c apaf 1 and caspase family members which participate in apoptosis pathways van gurp et al 2003 joza et al 2003 and galindo et al 2003 .  
15964487cytochrome ccytochrome c1.0the activity of complex ii_amp_#x2013;iii succinate cytochrome c reductase was determined following the method of king 1967 .  
15964487cytochrome ccytochrome c1.0indeed calcium induces the release of mitochondrial cytochrome c by different mechanisms selective for brain versus liver andreyev and fiskum 1999 .  
16026864cytochrome ccytochrome c1.0most interestingly recent work provided evidence that mutant sod1 might disrupt association of complex iv cytochrome c with the inner mitochondrial membrane and by this interfere with mitochondrial respiration [48] .  
16046141cytochrome ccytochrome c1.0vation of the caspases cascade is also effective in preventing both the mitochondrial damage and the increase in the production of reactive oxygen species induced by fals sod1 even in the presence of cytochrome c release.  
16046141cytochrome ccytochrome c1.0mitochondria alterations membrane depolarization decreased activity of respiratory complexes cytochrome c release and oxidative stress increased ros flux oxidatively modified proteins seem likely candidates to explain many facets of als because of their early occurrence in experimental models and in pati 
16046141cytochrome ccytochrome c1.0of apoptosis by oxidative stress is a feature observed in several experimental paradigms cai and jones 1998 and luetjens et al. 2000 and oxidative stress mediated mitochondrial dysfunction leading to cytochrome c release plays an important role in the initiation of motor neuron death by mutant sod1.  
16046141cytochrome ccytochrome c1.0in spinal cord specimens of both als patients and in the mice model for fals evidence of prominent recruitment of the mitochondrial apoptotic pathway has been documented by the observation of cytochrome c release and by the observation that prevention of cytochrome c release prolongs the lifespan of transgenic sod1 g93a mice zhu et al. 2002 .  
16046141cytochrome ccytochrome c1.0 release and by the observation that prevention of cytochrome c release prolongs the lifespan of transgenic sod1 g93a mice zhu et al. 2002 .  
16046141cytochrome ccytochrome c1.0upon release from mitochondria cytochrome c becomes part of the apoptosome a complex in which apaf1 serves as a scaffold protein also for the binding of pro caspase 9 the apoptosome recruits and processes caspase 9 to form a holoenzyme complex 
16046141cytochrome ccytochrome c1.0our data demonstrate that removal of apaf1 prevents cell death and mitochondrial damage by intercepting activation of the caspases cascade even in the presence of cytochrome c release.  
16046141cytochrome ccytochrome c1.0for cytochrome c and aif release experiments etna cells were harvested in hypotonic buffer 2 mm mgcl 2 10 mm kcl 10 mm tris_amp_#x2013;hcl ph 7.6 supplemented with protease inhibitor cocktail sigma and incubated for  
16046141cytochrome ccytochrome c1.0mouse monoclonal anti cytochrome c antibody was purchased from bd bioscience.  
16046141cytochrome ccytochrome c1.0an anti cytochrome c monoclonal antibody clone 6h2.b4 pharmingen an anti cytochrome c polyclonal antibody santa cruz an anti aif monoclonal antibody clone e1 santa cruz and an anti endog polyclonal antibody prosci incorporated were used.  
16046141cytochrome ccytochrome c1.0cytochrome c release induced by fals sod1 is independent of apaf1 expression  
16046141cytochrome ccytochrome c1.0mitochondrial damage and cytochrome c release are pro apoptotic features observed in several experimental paradigms for fals see introduction .  
16046141cytochrome ccytochrome c1.0to compare cytochrome c release in cells expressing either fals sod1s or wild type sod1 etna cells were transfected with cdna coding for gfp sod1s fusion proteins.  
16046141cytochrome ccytochrome c1.0furthermore while no cytochrome c release is observed in cells expressing gfp or the wild type enzyme both etna_amp_#x2212;/_amp_#x2212; and etna+/+ cells expressing different fals sod1s display cytochrome c release independently from the apaf1 genotype figs 4 b_amp_#x2013;d .  
16046141cytochrome ccytochrome c1.0mitochondrial release of pro apoptotic factors induced by fals sod1 is specific for cytochrome c ; indeed we have observed that other mitochondrial factors involved in apoptotic pathways such as aif fig 4 and fig 5 and endog not shown are not affected by the expression of mutant sod1s in this sy 
16046141cytochrome ccytochrome c1.0mitochondrial membrane depolarization decreased activity of respiratory complexes and cytochrome c release occur at the asymptomatic stage in fals sod1 transgenic mice bendotti et al. 2001 and jung et al. 2002 and in cultured cells expressing the mutant protein carr_amp_#xec; et al. 1997 beal 2000 
16046141cytochrome ccytochrome c1.0respiratory chain leading to bioenergetic failure in the motor neurons bendotti et al. 2001 and jung et al. 2002 and to alteration in the mitochondrial permeability transition pore causing leakage of cytochrome c and apoptosis inducing factor aif friedlander 2003 were suggested to play a role in the mitochondrial dependent motor neuron death through the activation of a caspase mediated cascade leading to prog 
16046141cytochrome ccytochrome c1.01 is a key factor in the noxious function of fals sod1: while overexpression of apaf1 exacerbates the induction of apoptosis fig 2 its removal prevents activation of caspase 3 even in the presence of cytochrome c release fig 3 and fig 4 .  
16046141cytochrome ccytochrome c1.0either exert some aberrant chemistry specifically inside or on the surface of mitochondria or interfere with cytoplasmic functions modulating mitochondria functionality: both could lead to release of cytochrome c activation of the caspase cascade and impairment in the respiratory chain atp depletion and changes in the membrane permeability.  
16046141cytochrome ccytochrome c1.0in a recent paper kirkinezos et al. have reported that cytochrome c association with the inner mitochondrial membrane is impaired in the cns of g93a sod1 mice kirkinezos et al. 2005 .  
16046141cytochrome ccytochrome c1.0gfp green cytochrome c red and merged patterns of the same representative fields are shown.  
16046141cytochrome ccytochrome c1.0 c the proportion of gfp positive cells releasing cytochrome c was scored n _amp_#xa0;=_amp_#xa0;3 .  
16046141cytochrome ccytochrome c1.0 d cytosolic fractions of etna cells transfected for 24 h with wtsod1 or g93a g37r g85r and i113t sod1 mutants were assessed for the presence of cytochrome c and aif by western blot.  
16046141cytochrome ccytochrome c1.0extracts from the mitochondrial fraction of untransfected etna cells were used as a positive control for aif and cytochrome c expression.  
16046141cytochrome ccytochrome c1.0double labeling confocal immunofluorescence microscopy staining of cytochrome c green and aif red in etna cells 48 h after transfection with wtsod1 gfp g93a sod1 gfp g85r sod1 gfp and a4v sod1 gfp.  
16046141cytochrome ccytochrome c1.0white arrowheads point to cells with released cytochrome c .  
16050975cytochrome ccytochrome c1.0in spinal motor neurons of g93a transgenic mice cytochrome c is released from mitochondria leading to caspase 9 activation guegan et al. 2001 .  
16050975cytochrome ccytochrome c1.0one may envision a scenario where mitochondrial dysfunction results in quantal releases of pro apoptotic factors such as cytochrome c apoptosis inducing factor aif and endog from individual mitochondria perhaps in response to local calcium mediated toxicity for example under excitatory synapses.  
16050975cytochrome ccytochrome c1.0higgins and colleagues have observed that sod1 and cytochrome c a resident protein of the intermembrane space colocalize at the early stages of mitochondrial vacuolization in mutant sod1 transgenic mice.  
16050975cytochrome ccytochrome c1.0the expanded outer membrane could become porous and allow for leakage of cytochrome c and other pro apoptotic molecules into the cytosol potentially triggering the apoptotic cascade higgins et al. 2003 and xu et al. 2004 .  
16242643cytochrome ccytochrome c1.0this blockade in turn inhibits the release of the proapoptotic factors cytochrome c smac diablo and apoptosis inducing factor from the mitochondria wang et al. 2003 and zhu et al. 2002 .  
16242643cytochrome ccytochrome c1.0cardiolipin is a phospholipid located in the inner mitochondrial membrane whose oxidation can be the consequence of the release of cytochrome c from the mitochondria to the cytoplasm tuominen et al. 2002 a release that we have previously observed to be increased in sh sy5y cells challenged with malonate fern_amp_#xe1;ndez g_amp_#xf3;mez et a 
16242643cytochrome ccytochrome c1.0as a consequence of permeability transition pore formation cytochrome c is released to the cytoplasm turning on the apoptotic machinery.  
17105868cytochrome ccytochrome c1.0altered expression and dysfunction of bcl 2 may contribute to the activation of mitochondrial apoptosis machinery such as caspase 9 caspase 3 and cytochrome c in spinal motor neurons of als transgenic mice and humans with als guegan et al. 2001 ; inoue et al. 2003 .  
17150307cytochrome ccytochrome c1.0furthermore depletion of gsh decreased mitochondrial function facilitated apoptosis inducing factor aif translocation cytochrome c release and caspase 3 activation and consequently led to motor neuron like cell apoptosis.  
17150307cytochrome ccytochrome c1.0after protein concentration measurement western blotting was performed to analyze cytochrome c release.  
17150307cytochrome ccytochrome c1.0western analysis was performed to analyze cytochrome c release and aif translocation.  
17150307cytochrome ccytochrome c1.0after permeabilization with 0.2% triton x 100 in pbs and blocking with 10% goat serum cells were incubated with specific antibodies overnight at 4 _amp_#x000b0;c aif cytochrome c and active caspase 3 antibodies were all at 1:300 dilution; chemicon inc. .  
17150307cytochrome ccytochrome c1.0results gsh depletion promoted aif redistribution cytochrome c release caspase 3 activation and apoptotic cell death a biochemical assay and an immunohistochemical analysis were used to identify the molecular pathways of cell apoptosis by gsh depletion.  
17150307cytochrome ccytochrome c1.0similarly mitochondrial and cytosolic fractionation coupled with western blotting analysis and immunohistochemistry showed that cytochrome c release was involved in nsc34 cell apoptosis caused by gsh depletion figure 6c 6d .  
17150307cytochrome ccytochrome c1.0cytochrome c mediated caspase 3 activation elicited by gsh reduction also contributed to motor neuron cell death apoptosis .  
17150307cytochrome ccytochrome c1.0the dramatic increase in cytochrome c release to cytosol detected by the fractionation assay and immunohistochemical analysis figure 6b and 6d most likely reflects the increased activation of caspase 3 and consequently contributes to the 
17150307cytochrome ccytochrome c1.0cytochrome c mediated caspase activation has also been shown to lead motor neuron degeneration in als like mouse and human als patients guegan et al. 2001 ; li et al. 2000 .  
17496232cytochrome ccytochrome c1.0increased no and onoo result in dna damage p53 accumulation increased bax/bcl 2 cytochrome c cyt c release and caspase activation.  
17496232cytochrome ccytochrome c1.0the proapoptotic effects depend on translocation of bax to mitochondria and generation of ros and ultimately on the release of cytochrome c but in a caspase independent manner.  
17496232cytochrome ccytochrome c1.0in the intrinsic program mitochondrial damage results in the release of cytochrome c triggering the assembly of the apoptosome complex with apoptotic protease activating factor 1 as central scaffold protein that directly recruits caspase 9 which in turn elicits caspase 3 effects 31 . 
17496232cytochrome ccytochrome c1.0no can directly induce cytochrome c release through mitochondrial membrane potential loss 46 or by tyrosine nitration of cytochrome c 67 .  
17496232cytochrome ccytochrome c1.0no mediated p53 accumulation induces cell cycle arrest by p21 upregulation or apoptosis by increase in bax/bcl xl cytochrome c release and caspase activation 100 .  
17496232cytochrome ccytochrome c1.0on the other hand no has antiapoptotic effects that can be associated with cgmp production which suppresses mitochondrial cytochrome c release ceramide generation and caspase activation 46 77 .  
17634371cytochrome ccytochrome c1.0in the present study we show that p75 mediated apoptosis in motor neurons involved neutral sphingomyelinase activation increased mitochondrial superoxide production and cytochrome c release to the cytosol.  
17634371cytochrome ccytochrome c1.0the death pathway is not completely elucidated but is associated with activation of c jun n terminal kinase jnk release of cytochrome c from mitochondria and subsequent activation of caspases 9 6 and 3 casaccia bonnefil et al. 1996 ; yoon et al. 1998 ; wang et al. 2001 ; bhakar et al. 2003 .  
17634371cytochrome ccytochrome c1.0ceramide could be a mediator of apoptosis by acting directly or indirectly on mitochondria where it can dissipate the membrane potential promote cytochrome c release and induce reactive oxygen species ros production garcia ruiz et al. 1997 ; gudz et al. 1997 ; quillet mary et al. 1997 ; mansat de mas et al. 1999 ; birbes et al. 2002 .  
17634371cytochrome ccytochrome c1.0after 12 h cultures were fixed as described above and processed for cytochrome c immunolabeling using alexa fluor 488 cytochrome c apoptosis detection kit from invitrogen.  
17634371cytochrome ccytochrome c1.0in the present study we found that the p75 apoptotic pathway in motor neurons involved increased ceramide production and cytochrome c release into the cytosol as observed in other cell types casaccia bonnefil et al. 1996 ; kuner and hertel 1998 ; brann et al. 2002 ; bhakar et al. 2003 . p75 induced ceramide generation in motor neur 
17634371cytochrome ccytochrome c1.0it is noteworthy that the increased superoxide production by mitochondria does not require nitric oxide whereas cytochrome c release does require the presence of an external source of nitric oxide such as the addition of deta nonoate.  
17634371cytochrome ccytochrome c1.0these results suggest that peroxynitrite a strong oxidant formed from the reaction of superoxide and nitric oxide disrupts mitochondrial function to promote cytochrome c release beckman and koppenol 1996 ; radi et al. 2002 .  
17634371cytochrome ccytochrome c1.0ngf induced apoptosis in motor neurons involves nsmase activation and triggers cytochrome c release from mitochondria.  
17634371cytochrome ccytochrome c1.0b fluorescence microphotographs showing cytochrome c immunoreactivity in motor neurons maintained with gdnf control or exposed to ngf plus deta nonoate ngf+no .  
17634371cytochrome ccytochrome c1.0in the absence of ngf motor neurons showed a punctuate mitochondrial labeling of cytochrome c whereas 12 h after treatment with ngf+no motor neurons showed a diffuse cytoplasmic labeling.  
17634371cytochrome ccytochrome c1.0c gw4869 100 n m ; gw prevented cytochrome c release induced by ngf+no.  
17634371cytochrome ccytochrome c1.0gw alone did not affect cytochrome c labeling data not shown . * p _lt_ 0.05 significantly different from control.  
17634371cytochrome ccytochrome c1.0c fluorescence microphotographs showing cytochrome c immunoreactivity in sod1 motor neurons maintained with gdnf and exposed to vehicle control or ngf 100 ng/ml .  
17634371cytochrome ccytochrome c1.0in the absence of ngf motor neurons showed a punctuate labeling of cytochrome c and 12 h after treatment with ngf a diffuse labeling was observed in affected motor neurons.  
17634371cytochrome ccytochrome c1.0immunofluorescence studies revealed that nsmase activation was followed by cytochrome c release from mitochondria fig 1 b .  
17634371cytochrome ccytochrome c1.0in the absence of ngf motor neurons showed a punctate pattern of cytochrome c immunoreactivity indicative of mitochondrial localization.  
17634371cytochrome ccytochrome c1.0ngf 100 ng/ml or deta nonoate 10 micro m treatment alone did not affect cytochrome c localization fig 1 c .  
17634371cytochrome ccytochrome c1.0in contrast after 12 h of ngf treatment in the presence of deta nonoate ~27% of motor neurons displayed a diffuse pattern of cytochrome c immunoreactivity indicating release into the cytoplasm fig 1 b c .  
17634371cytochrome ccytochrome c1.0cytochrome c release induced by ngf in the presence of nitric oxide was prevented by the addition of the nsmase inhibitor gw4869 fig 1 c indicating that cytochrome c release required ceramide production.  
17634371cytochrome ccytochrome c1.0moreover p75 mediated apoptosis in sod1 motor neurons was prevented by nsmase inhibitors fig 3 b and involved mitochondrial production and cytochrome c release fig 3 c d .  
18210200cytochrome ccytochrome c1.0for example a recent study reported a nanotube produced by a layer based assembly of cytochrome c poly sodium styrene sulfonate and poly ethylenimine pei .