scholarly journals ERK activation promotes neuronal degeneration predominantly through plasma membrane damage and independently of caspase-3

2004 ◽  
Vol 165 (3) ◽  
pp. 357-369 ◽  
Author(s):  
Srinivasa Subramaniam ◽  
Ute Zirrgiebel ◽  
Oliver von Bohlen und Halbach ◽  
Jens Strelau ◽  
Christine Laliberté ◽  
...  
Keyword(s):  
Dna Damage ◽  
Plasma Membrane ◽  
Protein Kinase ◽  
Caspase 3 ◽  
Neuronal Degeneration ◽  
Neuronal Damage ◽  
Membrane Damage ◽  
Cellular Damage ◽  
Nuclear Condensation ◽  
Erk Activation

Our recent studies have shown that extracellular-regulated protein kinase (ERK) promotes cell death in cerebellar granule neurons (CGN) cultured in low potassium. Here we report that the “death” phenotypes of CGN after potassium withdrawal are heterogeneous, allowing the distinction between plasma membrane (PM)–, DNA-, and PM/DNA-damaged populations. These damaged neurons display nuclear condensation that precedes PM or DNA damage. Inhibition of ERK activation either by U0126 or by dominant-negative mitogen-activated protein kinase/ERK kinase (MEK) overexpression results in a dramatic reduction of PM damaged neurons and nuclear condensation. In contrast, overexpression of constitutively active MEK potentiates PM damage and nuclear condensation. ERK-promoted cellular damage is independent of caspase-3. Persistent active ERK translocates to the nucleus, whereas caspase-3 remains in the cytoplasm. Antioxidants that reduced ERK activation and PM damage showed no effect on caspase-3 activation or DNA damage. These data identify ERK as an important executor of neuronal damage involving a caspase-3–independent mechanism.

scholarly journals Correction: ERK activation promotes neuronal degeneration predominantly through plasma membrane damage and independently of caspase-3

2004 ◽  
Vol 165 (4) ◽  
pp. 593-593
Author(s):  
Srinivasa Subramaniam ◽  
Ute Zirrgiebel ◽  
Oliver von Bohlen und Halbach ◽  
Jens Strelau ◽  
Christine Laliberté ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Caspase 3 ◽  
Neuronal Degeneration ◽  
Membrane Damage ◽  
Erk Activation ◽  
Plasma Membrane Damage

scholarly journals Mechanism of Chronic Stress-Induced Glutamatergic Neuronal Damage in the Basolateral Amygdaloid Nucleus

2021 ◽  
Vol 2021 ◽  
pp. 1-14
Author(s):  
Songjun Wang ◽  
Xia Liu ◽  
Weibo Shi ◽  
Qian Qi ◽  
Guozhong Zhang ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Behavioral Disorders ◽  
Chronic Stress ◽  
Structural Changes ◽  
Neuronal Degeneration ◽  
Neuronal Damage ◽  
Amygdaloid Nucleus ◽  
Protein Kinase R ◽  
Functional Changes ◽  
Glutamatergic Neurons

Stress is a ubiquitous part of our life, while appropriate stress levels can help improve the body’s adaptability to the environment. However, sustained and excessive levels of stress can lead to the occurrence of multiple devastating diseases. As an emotional center, the amygdala plays a key role in the regulation of stress-induced psycho-behavioral disorders. The structural changes in the amygdala have been shown to affect its functional characteristics. The amygdala-related neurotransmitter imbalance is closely related to psychobehavioral abnormalities. However, the mechanism of structural and functional changes of glutamatergic neurons in the amygdala induced by stress has not been fully elucidated. Here, we identified that chronic stress could lead to the degeneration and death of glutamatergic neurons in the lateral amygdaloid nucleus, resulting in neuroendocrine and psychobehavioral disorders. Therefore, our studies further suggest that the Protein Kinase R-like ER Kinase (PERK) pathway may be therapeutically targeted as one of the key mechanisms of stress-induced glutamatergic neuronal degeneration and death in the amygdala.

Abstract 204: Protective Roles of Eukaryotic Elongation Factor 2 Kinase on Rat Cardiomyoblast Death Under Glucose-deprived Condition

2017 ◽  
Vol 121 (suppl_1) ◽  
Author(s):  
Satoshi Kameshima ◽  
Muneyoshi Okada ◽  
Hideyuki Yamawaki
Keyword(s):  
Cell Death ◽  
Protein Kinase ◽  
Caspase 3 ◽  
Elongation Factor ◽  
H9c2 Cell ◽  
H9c2 Cells ◽  
Nuclear Condensation ◽  
Elongation Factor 2 ◽  
Eukaryotic Elongation Factor 2 ◽  
Eukaryotic Elongation Factor

Eukaryotic elongation factor 2 (eEF2) kinase (eEF2K; also known as calmodulin-dependent protein kinase III) has both active and inactive phosphorylation sites. An intracellular energy sensor, AMP-activated protein kinase (AMPK) was reported to activate eEF2K via increasing dephosphorylation at Ser366 (inactive site). Activated eEF2K phosphorylates and inactivates a specific substrate, eEF2, which results in the inhibition of protein translation consuming high energy. Glucose depletion (GD) is one of the primary causes for cardiomyocyte death in the developed cardiac hypertrophy. We have recently found that the expression and dephosphorylation of eEF2K (Ser366) and eEF2 phosphorylation were significantly increased in left ventricle of several cardiac hypertrophy models. However, it is almost unknown whether eEF2K/eEF2 signals affect GD-induced cardiomyocyte death. The aim of this study was to explore it. GD was induced by incubating H9c2 cells in a glucose-free medium. H9c2 cell viability, apoptotic-like nuclear condensation or protein expression was examined using a cell counting assay, DAPI staining or Western blotting, respectively. GD induced H9c2 cell death (p<0.01, n=6) and caspase-3 fragmentation (p<0.05, n=10-12). In addition, GD significantly increased phosphorylation of AMPK (p<0.05, n=6-8) and eEF2 (p<0.01, n=4-8) as well as eEF2K dephosphorylation at Ser366 (p<0.01, n=4-8). eEF2K gene knockdown (eEF2K KD) by siRNA transfection significantly increased GD-induced H9c2 cell death (p<0.05, n=7) and caspase-3 fragmentation (p<0.01, n=9). Moreover, eEF2K KD significantly facilitated GD-induced increase of nuclear condensation (44.0±3.3%, eEF2K siRNA vs. 30.9±2.4%, control siRNA p<0.01, n=5). AMPK KD did not affect GD-induced H9c2 cell death and eEF2K dephosphorylation. In conclusion, we for the first time revealed in H9c2 cells that activated eEF2K might play protective roles in GD-induced apoptosis via the inhibition of caspase-3 fragmentation, whereas AMPK activation is not directly related to the regulation of eEF2K/eEF2 signals in GD condition. The present results suggest eEF2K as a novel pharmacotherapeutic target for cardiac dysfunction.

Role of cytosolic Ca in renal tubule damage induced by anoxia

1991 ◽  
Vol 260 (3) ◽  
pp. C545-C554 ◽  
Author(s):  
W. R. Jacobs ◽  
M. Sgambati ◽  
G. Gomez ◽  
P. Vilaro ◽  
M. Higdon ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Damage ◽  
Cellular Damage ◽  
Rabbit Kidney ◽  
Renal Tubules ◽  
Plasma Membrane Permeability ◽  
Fura 2 ◽  
Chemical Hypoxia ◽  
Independent Events ◽  
Energy Deprivation

Cytosolic free Ca (Caf) was measured in three different preparations of freshly prepared proximal tubules from the rabbit kidney during energy deprivation using fura-2. Isolated perfused tubules, tubules immobilized on glass cover slips, and tubules in suspension were subjected to inhibitors of oxidative phosphorylation (“chemical hypoxia”); the latter two preparations were also subjected to 40 min of anoxia. During normoxia, Caf ranged from 100 to 180 nM in all three preparations, and chemical hypoxia caused either no change or a small (30-100%) increase in Caf values. Subsequent addition of Ca ionophores increased Caf to 300-500 nM in the first 2 min and to greater than 1 microM after 15 min. In individual experiments, anoxia produced similar responses to those of chemical hypoxia, eliciting no average significant change in Caf, despite clear evidence for impaired respiration and plasma membrane damage after 40 min of anoxia. This lack of change in Caf was unrelated to “Ca buffering” by fura-2 or inactivation of the dye, since Caf increased to 666 +/- 59 nM upon addition of Ca ionophore during anoxia. These data suggest that increased Caf is not a prerequisite for cellular damage during anoxia in proximal renal tubules. Furthermore, no apparent alteration in plasma membrane permeability to Ca occurs before membrane disruption. Decreased ATP seems to initiate a series of Caf-independent events that cause irreversible injury.

Cytotoxicity and Genotoxic Effects of Organic Extracts from Traffic-Related Particulate Matter in Human Vascular Endothelial Cells

2012 ◽  
Vol 610-613 ◽  
pp. 686-690
Author(s):  
Yu Shang ◽  
Lan Lan Fan ◽  
Ling Zhang
Keyword(s):  
Oxidative Stress ◽  
Dna Damage ◽  
Endothelial Cells ◽  
Plasma Membrane ◽  
Particulate Matter ◽  
Vascular Endothelial Cells ◽  
Membrane Damage ◽  
Cell Culture Supernatant ◽  
Vascular Endothelial ◽  
Organic Extracts

Traffic-related particulate matter (PM) is associated with adverse cardiovascular effects. However, the direct impact of the traffic-related PM on the cytotoxicity and genotoxic damage in vascular endothelial cells is less well known. The aim of the present study is to assess whether exposure to the organic extracts of traffic related-PM (oTRP) can induce cytotoxicity, DNA damage and oxidative stress in the human umbilical vein endothelial cells (HUVEC). The cell viability was determined by MTT assay after treatment with oTRP for 24 h. The injury of the cell plasma membrane was evaluated by testing the lactate dehydrogenase (LDH) leakage in cell culture supernatant. The activity of superoxide dismutase (SOD) was determined to evaluate the level of oxidative stress in cells. DNA damage was measured by alkaline Comet assay. The results showed that significantly dose-dependent cytotoxicity and DNA damage was detected in HUVEC after treated with oTRP for 24 h. A concomitant decrease in SOD activity was observed, suggesting that oTRP may mediate genotoxic and cytotoxic effects in HUVEC through the oxidative stress pathway. These results indicate that organic extracts of traffic related-PM has a potency to cause oxidative stress, DNA damage, cell death and plasma membrane damage in HUVEC, thus may be participated in the development of cardiovascular diseases through damaging vascular endothelial cells.

scholarly journals Chemopreventive Effect of β-Cryptoxanthin on Human Cervical Carcinoma (HeLa) Cells Is Modulated through Oxidative Stress-Induced Apoptosis

Antioxidants ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 28 ◽  
Author(s):  
Enkhtaivan Gansukh ◽  
Arti Nile ◽  
Iyyakkannu Sivanesan ◽  
Kannan R. R. Rengasamy ◽  
Doo-Hwan Kim ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Dna Damage ◽  
Cervical Carcinoma ◽  
Hela Cells ◽  
Caspase 3 ◽  
Nuclear Condensation ◽  
Physiological Concentration ◽  
Human Cervical Carcinoma ◽  
Induced Apoptosis

The present study was aimed to assess cellular and molecular events involved in the chemopreventive activities of β-cryptoxanthin derived from mandarin oranges (Citrus unshiu Marc.) on human cervical carcinoma (HeLa) cells. In vitro experiments established that β-cryptoxanthin significantly inhibited the proliferation of HeLa cells with the IC50 value of 4.5 and 3.7 µM after 24 and 48 h of treatments, respectively. β-cryptoxanthin-treated HeLa cells exhibited enhanced levels of oxidative stress correlated with significant downregulation of anti-apoptotic Bcl-2, and upregulation of pro-apoptotic Bax mRNA expression. Moreover, β-cryptoxanthin triggered nuclear condensation and disruption of the integrity of the mitochondrial membrane, upregulated caspase-3, -7, and -9 mRNA, and enhanced activation of caspase-3 proteins, resulting in nuclei DNA damage and apoptosis of HeLa cells. Remarkably, TUNEL assay carried out to detect nuclei DNA damage showed 52% TUNEL-positive cells after treatment with a physiological concentration of β-cryptoxanthin (1.0 μM), which validates its potential as an anticancer drug of natural origin.

Protein kinase Cδ-dependent and -independent signaling in genotoxic response to treatment of desferroxamine, a hypoxia-mimetic agent

2007 ◽  
Vol 292 (6) ◽  
pp. C2150-C2160 ◽  
Author(s):  
Carlos Clavijo ◽  
Jo-Lin Chen ◽  
Kwang-Jin Kim ◽  
Mary E. Reyland ◽  
David K. Ann
Keyword(s):  
Dna Damage ◽  
Protein Kinase ◽  
Caspase 3 ◽  
Fluorescent Protein ◽  
Critical Role ◽  
Dominant Negative ◽  
Fine Tuning ◽  
Time Dependent ◽  
Response To Treatment ◽  
Dependent Manner

Protein kinase C (PKC) plays a critical role in diseases such as cancer, stroke, and cardiac ischemia and participates in a variety of signal transduction pathways including apoptosis, cell proliferation, and tumor suppression. Here, we demonstrate that PKCδ is proteolytically cleaved and translocated to the nucleus in a time-dependent manner on treatment of desferroxamine (DFO), a hypoxia-mimetic agent. Specific knockdown of the endogenous PKCδ by RNAi (sh-PKCδ) or expression of the kinase-dead (Lys376Arg) mutant of PKCδ (PKCδKD) conferred modulation on the cellular adaptive responses to DFO treatment. Notably, the time-dependent accumulation of DFO-induced phosphorylation of Ser-139-H2AX (γ-H2AX), a hallmark for DNA damage, was altered by sh-PKCδ, and sh-PKCδ completely abrogated the activation of caspase-3 in DFO-treated cells. Expression of Lys376Arg-mutated PKCδ-enhanced green fluorescent protein (EGFP) appears to abrogate DFO/hypoxia-induced activation of endogenous PKCδ and caspase-3, suggesting that PKCδKD-EGFP serves a dominant-negative function. Additionally, DFO treatment also led to the activation of Chk1, p53, and Akt, where DFO-induced activation of p53, Chk1, and Akt occurred in both PKCδ-dependent and -independent manners. In summary, these findings suggest that the activation of a PKCδ-mediated signaling network is one of the critical contributing factors involved in fine-tuning of the DNA damage response to DFO treatment.

Anticancer Activity of Platinum (II) Complex with 2-Benzoylpyridine by Induction of DNA Damage, S-Phase Arrest, and Apoptosis

2020 ◽  
Vol 20 (4) ◽  
pp. 504-517
Author(s):  
Yu-Lan Li ◽  
Xin-Li Gan ◽  
Rong-Ping Zhu ◽  
Xuehong Wang ◽  
Duan-Fang Liao ◽  
...  
Keyword(s):  
Dna Damage ◽  
B Cell ◽  
Anticancer Activity ◽  
Cancer Cells ◽  
Hepg2 Cells ◽  
Caspase 3 ◽  
Cell Lymphoma ◽  
B Cell Lymphoma ◽  
S Phase ◽  
Caspase 9

Objective: To overcome the disadvantages of cisplatin, numerous platinum (Pt) complexes have been prepared. However, the anticancer activity and mechanism of Pt(II) complexed with 2-benzoylpyridine [Pt(II)- Bpy]: [PtCl2(DMSO)L] (DMSO = dimethyl sulfoxide, L = 2-benzoylpyridine) in cancer cells remain unknown. Methods: Pt(II)-Bpy was synthesized and characterized by spectrum analysis. Its anticancer activity and underlying mechanisms were demonstrated at the cellular, molecular, and in vivo levels. Results: Pt(II)-Bpy inhibited tumor cell growth, especially HepG2 human liver cancer cells, with a halfmaximal inhibitory concentration of 9.8±0.5μM, but with low toxicity in HL-7702 normal liver cells. Pt(II)- Bpy induced DNA damage, which was demonstrated through a marked increase in the expression of cleavedpoly (ADP ribose) polymerase (PARP) and gamma-H2A histone family member X and a decrease in PARP expression. The interaction of Pt(II)-Bpy with DNA at the molecular level was most likely through an intercalation mechanism, which might be evidence of DNA damage. Pt(II)-Bpy initiated cell cycle arrest at the S phase in HepG2 cells. It also caused severe loss of the mitochondrial membrane potential; a decrease in the expression of caspase-9 and caspase-3; an increase in reactive oxygen species levels; the release of cytochrome c and apoptotic protease activation factor; and the activation of caspase-9 and caspase-3 in HepG2 cells, which in turn resulted in apoptosis. Meanwhile, changes in p53 and related proteins were observed including the upregulation of p53, the phosphorylation of p53, p21, B-cell lymphoma-2-associated X protein, and NOXA; and the downregulation of B-cell lymphoma 2. Moreover, Pt(II)-Bpy displayed marked inhibitory effects on tumor growth in the HepG2 nude mouse model. Conclusion: Pt(II)-Bpy is a potential candidate for cancer chemotherapy.

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