Regulation of X-Chromosome-Linked Inhibitor of Apoptosis Protein in Kainic Acid-Induced Neuronal Death in the Rat Hippocampus

doi:10.1006/mcne.2000.0935

Molecular and Cellular Neuroscience

Volume 17, Issue 2, February 2001, Pages 364-372

https://doi.org/10.1006/mcne.2000.0935 Get rights and content

Abstract

XIAP (X-chromosome-linked inhibitor of apoptosis protein) is an antiapoptotic protein which inhibits the activity of caspases and suppresses cell death. However, little is known about the presence and function of XIAP in the nervous system. Here we report that XIAP mRNA is expressed in developing and adult rat brain. Using a specific antibody, we observed XIAP-immunoreactive cells in different brain regions, among others, in the hippocampus and cerebral cortex. Kainic acid, which induces delayed cell death of specific neurons, increased the levels of XIAP in the CA3 region of hippocampus. XIAP was, however, largely absent in cells undergoing cell death, as shown by TUNEL labeling and staining for active caspase-3. In cultured hippocampal neurons, XIAP was initially upregulated by kainic acid and then degraded in a process blocked by the caspase-3 inhibitor DEVD. Similarly, recombinant XIAP is cleaved by active caspase-3 in vitro. The results show that there is biphasic regulation of XIAP in the hippocampus following kainic acid and that XIAP becomes a target for caspase-3 activated during cell death in the hippocampus. The degradation of XIAP by kainic acid contributes to neuronal cell death observed in vulnerable neurons of the hippocampus after caspase activation.

References (37)

P. Chomczynski et al.
Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction
Anal. Biochem.
(1987)
P. Ernfors et al.
Increased levels of messenger RNAs for neurotrophic factors in the brain during kindling epileptogenesis
Neuron
(1991)
C.J. Faherty et al.
Caspase-3-dependent neuronal death in the hippocampus following kainic acid treatment
Mol. Brain Res.
(1999)
B.A. Hay et al.
Drosophila homologs of baculovirus inhibitor of apoptosis proteins function to block cell death
Cell
(1995)
T.H. Murphy et al.
L-type voltage-sensitive calcium channels mediate synaptic activation of immediate early genes
Neuron
(1991)
J.W. Olney et al.
Kainic acid: a powerful neurotoxic analog of glutamate
Brain Res.
(1974)
H. Pollard et al.
Kainate-induced apoptotic cell death in hippocampal neurons
Neuroscience
(1994)
M. Rothe et al.
The TNFR2-TRAF signalling complex contains two novel proteins related to baculovirus inhibitor of apoptosis
Cell
(1995)
N. Roy et al.
The gene for neuronal apoptosis inhibitory protein is partially deleted in individuals with spinal muscular atrophy
Cell
(1995)
T. Salin et al.
Up-regulation of trkB mRNA expression in the rat striatum after seizures
Neurosci. Lett.
(1995)

N.A. Simonian et al.

Kainic acid induces apoptosis in neurons

Neuroscience

(1996)

Y. Skoglösa et al.

Regulation of pituitary adenylate cyclase activating polypeptide and its receptor type 1 after traumatic brain injury: comparison with brain-derived neurotrophic factor and the induction of neuronal cell death

Neuroscience

(1999)

R. Takahashi et al.

A single BIR domain of XIAP sufficient for inhibiting caspases

J. Biol. Chem.

(1998)

G. Ambrosini et al.

A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma

Nature Med.

(1997)

R.J. Clem et al.

Control of programmed cell death by the baculovirus genes p35 and iap

Mol. Cell. Biol.

(1994)

Q.L. Deveraux et al.

IAPs block apoptotic events induced by caspase-8 and cytochrome c by direct inhibition of distinct caspases

EMBO J.

(1998)

Q.L. Deveraux et al.

Cleavage of human inhibitor of apoptosis protein XIAP results in fragments with distinct specificities for caspases

EMBO J.

(1999)

C. Duckett et al.

A conserved family of cellular genes related to the baculovirus iap gene and encoding apoptosis inhibitors

EMBO J.

(1996)

Cited by (69)

Peroxisome proliferator-activated receptor-γ (PPARγ) agonist is neuroprotective and stimulates PGC-1α expression and CREB phosphorylation in human dopaminergic neurons
2016, Neuropharmacology
Mitochondrial dysfunction has been linked to several neurodegenerative diseases such as Parkinson's disease (PD). Peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) is a master gene for mitochondrial biogenesis and has been shown to be neuroprotective in models of PD. In this work we have studied the mechanisms by which peroxisome proliferator-activated receptor-γ (PPARγ) selective agonist N-(2-benzoylphenyl)-O-[2-(methyl-2-pyridinylamino)ethyl]-l-tyrosine hydrate (GW1929) acts on human dopaminergic neurons in culture. Data showed that GW1929 increased the viability of human dopaminergic neurons and protected them against oxidative stress induced by H₂O₂ and the mitochondrial toxin Rotenone. The enhanced resilience of the neurons was attributed to increased levels of mitochondrial antioxidants and of PGC-1α. GW1929 treatment further increased cell respiration, mitochondrial biogenesis and sirtuin-1 (SIRT1) expression in the human dopaminergic neurons. Phosphorylation of cAMP responsive element-binding protein (CREB) was also robustly increased in GW1929-treated cells. Together these results show that the PPARγ agonist GW1929 influences CREB signaling and PGC-1α activities in the human dopaminergic neurons contributing to an increased cell viability. This supports the view that drugs acting on the PPARγ-PGC-1α signaling in neurons may have beneficial effects in PD and possible also in other brain disorders.
Reciprocal regulation of very low density lipoprotein receptors (VLDLRs) in neurons by brain-derived neurotrophic factor (BDNF) and reelin: Involvement of the E3 LIGASE mylip/idol
2013, Journal of Biological Chemistry
BDNF positively influences various aspects of neuronal migration, maturation, and survival in the developing brain. Reelin in turn mediates inhibitory signals to migrating neuroblasts, which is crucial for brain development. The interplay between BDNF and Reelin signaling in neurodevelopment is not fully understood. We show here that BDNF increased the levels of the Reelin receptor (VLDL receptor (VLDLR)) in hippocampal neurons by increasing gene expression. In contrast, Reelin decreased VLDLRs, which was accompanied by an increase in the levels of the E3 ligase Mylip/Idol in neurons. Down-regulation of Mylip/Idol using shRNAs abrogated the decrease in VLDLRs induced by Reelin. These results show that VLDLRs are tightly regulated in hippocampal neurons by both transcriptional and post-transcriptional mechanisms. The regulation of VLDLR by BDNF and Reelin may affect the migration of neurons and contribute to neurodevelopmental disorders in the nervous system.
Background: VLDLR is a receptor for Reelin in brain neurons, but its mode of regulation is unclear.
Results: BDNF increases VLDLR gene expression in hippocampal neurons, whereas Reelin decreases the receptor post-transcriptionally.
Conclusion: Reelin down-regulates VLDLR via the E3 ubiquitin ligase Mylip/Idol.
Significance: The dynamic regulation of VLDLR by BDNF and Reelin may be important for neuronal migration and brain development.
Fibroblast growth factor-21 (FGF21) regulates low-density lipoprotein receptor (LDLR) levels in cells via the E3-ubiquitin ligase Mylip/Idol and the Canopy2 (Cnpy2)/Mylip-interacting saposin-like protein (Msap)
2012, Journal of Biological Chemistry
The LDLR is a critical factor in the regulation of blood cholesterol levels that are altered in different human diseases. The level of LDLR in the cell is regulated by both transcriptional and post-transcriptional events. The E3 ubiquitin ligase, myosin regulatory light chain-interacting protein (Mylip)/inducible degrader of the LDL-R (Idol) was shown to induce degradation of LDLR via protein ubiquitination. We have here studied novel factors and mechanisms that may regulate Mylip/Idol in human hepatocyte cells and in mouse macrophages. We observed that FGF21 that is present in serum in different conditions reduced Mylip/Idol at the RNA and protein level, and increased LDLR levels and stability in the cells. FGF21 also enhanced expression of Canopy2 (Cnpy2)/MIR-interacting Saposin-like protein (Msap) that is known to interact with Mylip/Idol. Overexpression of Cnpy2/Msap increased LDLRs, and knockdown experiments showed that Cnpy2/Msap is crucial for the FGF21 effect on LDLRs. Experiments using DiI-labeled LDL particles showed that FGF21 increased lipoprotein uptake and the effect of FGF21 was additive to that of statins. Our results are consistent with an important role of FGF21 and Cnpy2/Msap in the regulation of LDLRs in cultured cells, which warrants further studies using human samples.
Endoplasmic reticulum stress contributes to CRH-induced hippocampal neuron apoptosis
2012, Experimental Cell Research
Citation Excerpt :
Pregnant Sprague–Dawley rats were supplied by the Experimental Animal Center of Hebei Medical University (Shijiazhuang, Hebei, China). Hippocampal neurons prepared from embryonic day 18 rats were cultured for 8 d on poly-l-lysine (0.1 mg/ml)-coated dishes in neurobasal medium with 2% B27 supplement as described previously [12,13]. Cultures were maintained at 37 °C in a humidified atmosphere of 95% air and 5% CO2.
The hypothalamic–pituitary–adrenal (HPA) axis is critical to mediating the body's response to stress. Corticotropin releasing hormone (CRH) plays a central role in controlling the stress response and regulating the HPA axis. Recent findings support CRH participates in the stress-induced hippocampal neuron apoptosis, but the underlying mechanisms are not fully understood. Our present study demonstrates that CRH can independently decrease hippocampal neuron cell viability in vitro in a concentration- and time-dependent manner. CRH receptor 1 (CRHR1) is involved in CRH-induced neuron apoptosis. Endoplasmic reticulum (ER) stress response marker, glucose-regulated protein 78 (GRP78), either protein or mRNA, is significantly elevated after treatment of CRH, and decreased when co-treated with salubrinal, ER stress inhibitor. The ER stress associated proapoptotic transcription factor C/EBP homologous protein (CHOP) and cleavage of caspase-12 protein expression are also increased following CRH treatment. Furthermore, we investigate which ER stress cascades are affected by CRH. CRH activates inositol-requiring enzyme 1 (IRE1), apoptosis signal regulating kinase 1 (ASK1), and c-jun kinase (JNK). Neuron apoptotic rate, examined by flow cytometry, is increased when CRH treatment and attenuated by salubrinal, thioredoxin (ASK1 inhibitor) and SP600125 (JNK inhibitor). Therefore, current data indicate that ER stress, through activating the IRE1/ASK1/JNK cascade, plays an important role in CRH-induced neuron apoptosis.
Involvement of estrogen receptors in the resveratrol-mediated increase in dopamine transporter in human dopaminergic neurons and in striatum of female mice
2012, Neuropharmacology
Citation Excerpt :
The films were developed in Kodak D19 developer and fixer (Eastman-Kodak). Immunoblots of cultured dopaminergic cells were performed essentially as described (Korhonen et al., 2001; Sokka et al., 2007; Kairisalo et al., 2011) and using primary antibodies against DAT (1:500; Novus Biologicals Littleton, CO, USA) and β-actin (1:5000). Appropriate peroxidase-conjugated antibodies (1:2500, Jackson ImmunoResearch, Biofellows, Helsinki, Finland) were added for 1 h and detection was performed using SuperSignal West Pico Substrate (Thermo).
Treatment with resveratrol (RSV) has been shown to protect vulnerable neurons after various brain injuries and in neurodegenerative diseases. The mechanisms for the effects of RSV in brain are not fully understood, but RSV may affect the expression of various gene products. RSV is structurally related to the synthetic estrogen, diethylstilbestrol so the effects of RSV may be gender-specific. Here we studied the role of RSV in the regulation of dopamine transporter (DAT) in the striatum using male and female mice. The basic levels of DAT in the striatum showed no sex difference, but the levels increased significantly by RSV (20 mg/kg i.p.) in female but not in male mice. Pretreatment of mice with the selective estrogen receptor (ER), ERα- and ERβ antagonist ICI 182,780, led to a complete block of RSV effect on DAT protein levels, suggesting that ERs are involved in the up-regulation of DAT by RSV. Similar data was also obtained in culture using human MESC2.10 and mouse SN4741 dopaminergic cells after treatment with RSV. Data further showed that RSV specifically induced gene transcription of DAT in the dopaminergic cells. These results show that estrogen receptors are involved in the up-regulation of DAT by RSV in the dopaminergic neurons, demonstrating a sex-dependent effect of RSV in the brain that may be of clinical importance.
This article is part of a Special Issue entitled ‘Post-Traumatic Stress Disorder’.
Expression of X-chromosome linked inhibitor of apoptosis protein in mature purkinje cells and in retinal bipolar cells in transgenic mice induces neurodegeneration
2008, Neuroscience
Citation Excerpt :
Cerebelli and eye bulbs from control and L7-XIAP mice were homogenized and protein lysates subjected to immunoblotting as described earlier. Primary antibodies were: anti-XIAP antibody (1:2000, BD Bioscience), anti-calbindinD (1:200, Cell Signaling), anti–protein kinase C-α (PKCα, 1:5000, Sigma), anti-p-c-Jun (1:1000, Cell Signaling), and β-actin (1:2000, Sigma) that was used as a control (Korhonen et al., 2001, 2005). Sections of 2-month-old cerebellum were immersion fixed with 1% paraformaldehyde and 2.5% glutaraldehyde overnight at room temperature, and postfixed for 1 h with 1% buffered osmium tetroxide.
Transgenic mice with overexpression of the caspase-inhibitor, X-chromosome-linked inhibitor of apoptosis protein (XIAP) in Purkinje cell (PC) and in retinal bipolar cells (RBCs) were produced to study the regulation of cell death. Unexpectedly, an increased neurodegeneration was observed in the PCs in these L7-XIAP mice after the third postnatal week with the mice exhibiting severe ataxia. The loss of PCs was independent of Bax as shown by crossing the L7-XIAP mice with Bax gene–deleted mice. Electron microscopy revealed intact organelles in PCs but with the stacking of ER cisterns indicative of cell stress. Immunostaining for cell death proteins showed an increased phosphorylation of c-Jun in the PCs, suggesting an involvement in cell degeneration. Apart from PCs, the number of RBCs was decreased in adult retina in line with the expression pattern for the L7 promoter. The data show that overexpression of the anti-apoptotic protein XIAP in vulnerable neurons leads to enhanced cell death. The mechanisms underlying this neurodegeneration can be related to the effects of XIAP on cell stress and altered cell signaling.

View all citing articles on Scopus

¹: To whom correspondence should be addressed at Department of Neuroscience, Neurobiology, Uppsala University, Biomedical Center, Box 587, S-75123 Uppsala, Sweden. Fax: ++46 18 559017. E-mail: [email protected].

View full text

Regular ArticleRegulation of X-Chromosome-Linked Inhibitor of Apoptosis Protein in Kainic Acid-Induced Neuronal Death in the Rat Hippocampus

Abstract

Anal. Biochem.

Neuron

Mol. Brain Res.

Cell

Neuron

Brain Res.

Neuroscience

Cell

Cell

Neurosci. Lett.

Neuroscience

Neuroscience

J. Biol. Chem.

A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma

Nature Med.

Control of programmed cell death by the baculovirus genes p35 and iap

Mol. Cell. Biol.

IAPs block apoptotic events induced by caspase-8 and cytochrome c by direct inhibition of distinct caspases

EMBO J.

Cleavage of human inhibitor of apoptosis protein XIAP results in fragments with distinct specificities for caspases

EMBO J.

A conserved family of cellular genes related to the baculovirus iap gene and encoding apoptosis inhibitors

EMBO J.

Regular Article
Regulation of X-Chromosome-Linked Inhibitor of Apoptosis Protein in Kainic Acid-Induced Neuronal Death in the Rat Hippocampus