Endogenous Bax Translocation in SH‐SY5Y Human Neuroblastoma Cells and Cerebellar Granule Neurons Undergoing Apoptosis

McGinnis, Kim M.; Gnegy, Margaret E.; Wang, Kevin

doi:10.1046/j.1471-4159.1999.0721899.x

Cited by 57 publications

(14 citation statements)

References 62 publications

(68 reference statements)

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“…In agreement with previous studies (Miller et al, 1997), treatment of CGNs with low K ϩ did not significantly modify the amount of total BAX measured by Western blot when compared with untreated cells (results not shown). Furthermore, consistent with previous results (McGinnis et al, 1999), biochemical experiments measuring the amount of total BAX in subcellular fractions showed that treatment of CGNs with low K ϩ increases the amount of BAX in mitochondria after 2 h of treatment (137 Ϯ 12% control) (Fig. 7D) but not after 1 h of treat- , and homozygote Bax-deficient (Bax Ϫ/Ϫ ) mice were treated with low K ϩ (K Ϫ ) or with control high K ϩ (K ϩ ) at DIV8.…”

Section: Np1 Regulates the Accumulation Of Bax In Mitochondriasupporting

confidence: 93%

NP1 Regulates Neuronal Activity-Dependent Accumulation of BAX in Mitochondria and Mitochondrial Dynamics

Clayton¹,

Podlesniy²,

Figueiro-Silva³

et al. 2012

J. Neurosci.

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In cultured cerebellar granule neurons, low neuronal activity triggers the intrinsic program of apoptosis, which requires protein synthesis-dependent BAX translocation to mitochondria, a process that may underlie neuronal damage in neurodegeneration. However, the mechanisms that link neuronal activity with the induction of the mitochondrial program of apoptosis remain unclear. Neuronal pentraxin 1 (NP1) is a pro-apoptotic protein induced by low neuronal activity that is increased in damaged neurites in Alzheimer's disease-affected brains. Here we report that NP1 facilitates the accumulation of BAX in mitochondria and regulates mitochondrial dynamics during apoptosis in rat and mouse cerebellar granule neurons in culture. Reduction of neuronal activity increases NP1 protein levels in mitochondria and contributes to mitochondrial fragmentation in a Bax-dependent manner. In addition, NP1 is involved in mitochondrial transport in healthy neurons. These results show that NP1 is targeted to mitochondria acting upstream of BAX and uncover a novel function for NP1 in the regulation of mitochondrial dynamics and trafficking during apoptotic neurodegeneration.

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Section: Np1 Regulates the Accumulation Of Bax In Mitochondriasupporting

confidence: 93%

NP1 Regulates Neuronal Activity-Dependent Accumulation of BAX in Mitochondria and Mitochondrial Dynamics

Clayton¹,

Podlesniy²,

Figueiro-Silva³

et al. 2012

J. Neurosci.

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“…Interestingly, the levels of µ-and m-calpain increased from DIV10 ( Figure 3B). These findings agree with the observed decrease in Bax and with the early cleavage of PARP, which preceded caspase-3 processing, because Bax, caspase-3, and PARP are proteolytically processed by calpains (Kobayashi et al, 1990;Shah et al, 1996;Wood et al, 1998;McGinnis et al, 1999b). Thus, we examined the levels of µ-and m-calpain.…”

Section: Patterns Of Apoptotic Molecules During Cultivationsupporting

confidence: 86%

Neuronal loss in primary long-term cortical culture involves neurodegeneration-like cell death via calpain and p35 processing, but not developmental apoptosis or aging

Kim

Oh²,

Park³

et al. 2007

Exp Mol Med

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Primary neuronal culture is a powerful tool to study neuronal development, aging, and degeneration. However, cultured neurons show signs of cell death after 2 or 3 weeks. Although the mechanism underlying this phenomenon has not been elucidated, several preventive methods have been identified. Here we show that the neuronal loss in primary cortical culture involves calpain activation and subsequent neuronal cell death. Neuronal loss during cultivation showed destruction of neurites and synapses, and a decrease in neuron numbers. µ-Calpain and m-calpain were initially activated and accumulated by increased RNA expression. This neuronal death exhibited neurodegenerative features, such as conversion of p35 to p25, which is important in the developmental process and in the pathogenesis of Alzheimer's disease. But, postnatal and aged rat cortex did not show calpain activation and prolonged processing of p35 to p25, in contrast to the long-term culture of cortical neurons. In addition, the inhibition of calpains by ALLM or ALLN blocked the conversion of p35 to p25, indicating that the calpain activity is essential for the neurodegenerative features of cell death. Taken together, this study shows that the neuronal loss in primary cortical cultures involves neurodegeneration-like cell death through the activation of calpains and the subsequent processing of p35 to p25, but not developmental apoptosis or aging. Our results suggest that the long term primary culture of cortical neurons represent a valuable model of neurodegeneration, such as Alzheimer's disease.

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“…In our studies, staurosporine induced transient hyperpolarization prior to cytochrome c release and caspase activation (caspase-3 and caspase-9) in SH-SY5Y cells expressing adenoviral constructs. This is in accordance with the previous studies and the finding that mitochondrial depolarization is not required for cytochrome c release to occur [Krohn et al, 1999].MOMP is also postulated to result from the formation of protein-permeable pores in the mitochondrial outer membrane, specifically by the Bcl-2 pro-apoptotic family member Bax [Liu et al, 1996;McGinnis et al, 1999;Pavlov et al, 2001;Wei et al, 2001;De Giorgi et al, 2002;Degli Esposti and Dive, 2003;Scorrano and Korsmeyer, 2003;Guo et al, 2004]. Cells treated with staurosporine displayed an increase in monomeric Bax and produced Bax dimers at mitochondria, suggesting that the mechanism for MOMP induced by staurosporine in SH-SY5Y cells is via protein-permeable pores.…”

supporting

confidence: 92%

“…MOMP is also postulated to result from the formation of protein-permeable pores in the mitochondrial outer membrane, specifically by the Bcl-2 pro-apoptotic family member Bax [Liu et al, 1996;McGinnis et al, 1999;Pavlov et al, 2001;Wei et al, 2001;De Giorgi et al, 2002;Degli Esposti and Dive, 2003;Scorrano and Korsmeyer, 2003;Guo et al, 2004]. Cells treated with staurosporine displayed an increase in monomeric Bax and produced Bax dimers at mitochondria, suggesting that the mechanism for MOMP induced by staurosporine in SH-SY5Y cells is via protein-permeable pores.…”

Section: Discussionmentioning

confidence: 99%

“…The Bcl-2 proapoptotic family member Bax which has been shown to oligomerize in the outer membrane in response to mitochondrial apoptotic stimuli has been postulated to form the pores [McGinnis et al, 1999;Degli Esposti and Dive, 2003;Scorrano and Korsmeyer, 2003]. Bax translocation and oligomerization in mitochondria in response to staurosporine treatment was evaluated.…”

Section: Mitochondrial-targeted Active Akt Decreases Bax Oligomerizatmentioning

confidence: 99%

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Mitochondrial‐targeted active Akt protects SH‐SY5Y neuroblastoma cells from staurosporine‐induced apoptotic cell death

Mookherjee

Quintanilla

Roh

et al. 2007

J of Cellular Biochemistry

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Akt is a serine/threonine protein kinase that plays a vital role in promoting cellular survival. Predominantly cytosolic, upon stimulation with growth-factors or stress, active Akt translocates into mitochondria, but the functions of Akt in mitochondria are not yet fully understood. Mitochondria play a central role in apoptotic pathways and given Akt's functions in the cytoplasm, Akt in mitochondria may help preserve mitochondrial integrity during cellular stress. To test if the translocation of Akt into mitochondria is neuroprotective, adenoviral vectors expressing a constitutively active Akt, Ad-HA-Akt (DD), and a constitutively active Akt with a mitochondrial targeting signal, Ad-Mito-HA-Akt (DD), were generated. Human SH-SY5Y neuroblastoma cells expressing the adenoviral constructs were treated with staurosporine to initiate intrinsic apoptotic cell death and several aspects of the mitochondrial apoptotic pathway were evaluated. Expression of active Akt targeted to mitochondria was found to be sufficient to significantly reduce staurosporineinduced activation of caspase-3 and caspase-9, the release of cytochrome c from mitochondria, and Bax oligomerization at mitochondria. These findings demonstrate that intramitochondrial active Akt results in efficient protection against apoptotic signaling. KeywordsAkt; mitochondria; apoptosis; caspase; Bax; Δψ m Akt (also known as protein kinase B) is a serine/threonine protein kinase that belongs to the AGC protein kinase subfamily that includes protein kinase A (PKA) and protein kinase C (PKC) [Peterson and Schreiber, 1999]. Akt is a predominant pro-survival protein that is regulated by phosphorylation. The phosphorylation of Akt is increased in response to activation of phosphoinositol 3-kinase (PI3K)-dependent pathways [Datta et al., 1996;Bellacosa et al., 1998]. Recruitment and activation of PI3K in response to specific ligands results in the generation of 3′-phosphoinositides at the plasma membrane and recruitment of Akt to the plasma membrane via its pleckstrin homology domain [Franke et al., 1997b]. Akt undergoes a conformational change and is subsequently activated by phosphorylation at either Thr 308 or Ser 473 , with maximal activity occurring when both sites are phosphorylated [Kohn et al., 1995;Alessi et al., 1996;Scheid et al., 2002a,b].There is unequivocal evidence that Akt is neuroprotective. For example, in experiments where cerebellar granule neurons were deprived of either serum or growth-factors, expression of wild- NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript type Akt enhanced survival while the expression of a dominant negative Akt mutant increased apoptosis . Similar studies conducted in sympathetic neurons also showed that Akt was necessary and sufficient for promoting cellular survival [Crowder and Freeman, 1998]. These and other studies demonstrate that activation of Akt protects against neuronal cell death in response to many different stressors [Goswami et al., 1999;Salinas et al., 2001;Yamaguchi et al., 2...

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Endogenous Bax Translocation in SH‐SY5Y Human Neuroblastoma Cells and Cerebellar Granule Neurons Undergoing Apoptosis

Cited by 57 publications

References 62 publications

NP1 Regulates Neuronal Activity-Dependent Accumulation of BAX in Mitochondria and Mitochondrial Dynamics

NP1 Regulates Neuronal Activity-Dependent Accumulation of BAX in Mitochondria and Mitochondrial Dynamics

Neuronal loss in primary long-term cortical culture involves neurodegeneration-like cell death via calpain and p35 processing, but not developmental apoptosis or aging

Mitochondrial‐targeted active Akt protects SH‐SY5Y neuroblastoma cells from staurosporine‐induced apoptotic cell death

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