Nicotine Suppresses Tunicamycin-Induced, But Not Thapsigargin-Induced, Expression of GRP78 during ER Stress-Mediated Apoptosis in PC12 Cells

Sasaya, Harue; Utsumi, Takahiro; Shimoke, Koji; Nakayama, Hitoshi; Matsumura, Yoshitaka; Fukunaga, Kenji; Ikeuchi, Toshihiko

doi:10.1093/jb/mvn063

Cited by 30 publications

(22 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The present study shows reduction of ER stress at nicotine concentrations 500-fold lower than those required to suppress UPR pathways in tunicamycin-treated PC12 cells, for which direct actions of nicotine on the ubiquitin-proteasome system were suggested (Sasaya et al, 2008). PC12 cells lack ␣4␤2 nAChRs but express lower-affinity ␣3␤4 nAChRs, which may be subject to chaperoning at relatively high nicotine concentrations.…”

Section: Upr Suppressed By Ligand-nachr Interactions In Er 767mentioning

confidence: 51%

Pharmacological Chaperoning of Nicotinic Acetylcholine Receptors Reduces the Endoplasmic Reticulum Stress Response

Srinivasan

Richards²,

Xia³

et al. 2012

Molecular Pharmacology

View full text Add to dashboard Cite

We report the first observation that endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) can decrease when a central nervous system drug acts as an intracellular pharmacological chaperone for its classic receptor. Transient expression of ␣4␤2 nicotinic receptors (nAChRs) in Neuro-2a cells induced the nuclear translocation of activating transcription factor 6 (ATF6), which is part of the UPR. Cells were exposed for 48 h to the full agonist nicotine, the partial agonist cytisine, or the competitive antagonist dihydro-␤-erythroidine; we also tested mutant nAChRs that readily exit the ER. Each of these four manipulations increased Sec24D-enhanced green fluorescent protein fluorescence of condensed ER exit sites and attenuated translocation of ATF6-enhanced green fluorescent protein to the nucleus. However, we found no correlation among the manipulations regarding other tested parameters [i.e., changes in nAChR stoichiometry (␣4 2 ␤2 3 versus ␣4 3 ␤2 2 ), changes in ER and trans-Golgi structures, or the degree of nAChR up-regulation at the plasma membrane]. The four manipulations activated 0 to 0.4% of nAChRs, which shows that activation of the nAChR channel did not underlie the reduced ER stress. Nicotine also attenuated endogenously expressed ATF6 translocation and phosphorylation of eukaryotic initiation factor 2␣ in mouse cortical neurons transfected with ␣4␤2 nAChRs. We conclude that, when nicotine accelerates ER export of ␣4␤2 nAChRs, this suppresses ER stress and the UPR. Suppression of a sustained UPR may explain the apparent neuroprotective effect that causes the inverse correlation between a person's history of tobacco use and susceptibility to developing Parkinson's disease. This suggests a novel mechanism for neuroprotection by nicotine.

show abstract

Section: Upr Suppressed By Ligand-nachr Interactions In Er 767mentioning

confidence: 51%

Pharmacological Chaperoning of Nicotinic Acetylcholine Receptors Reduces the Endoplasmic Reticulum Stress Response

Srinivasan

Richards²,

Xia³

et al. 2012

Molecular Pharmacology

View full text Add to dashboard Cite

show abstract

“…We found that treatment of PC12 cells with fisetin (up to 40 µM) alone for 16 h did not alter cell viability (Figure 1b). It is known that Tm blocks N -glycosylation of proteins, thus leading to ER stress [3] and ultimately apoptosis in PC12 cells [33,34]. Figure 1c shows that Tm (1–5 µg/mL) caused 30–40% PC12 cell death after 16 h. Treatment of PC12 cells with fisetin (5–20 µM) dose-dependently reversed 1 µg/mL Tm-mediated cell death (Figure 1d).…”

Section: Resultsmentioning

confidence: 95%

“…For experiments testing the ability of tunicamycin (Tm) to induce cytotoxicity, cells were incubated in serum-free RPMI medium in the presence of Tm [34]. PC12 cells (5 × 10 5 /mL) were seeded in 24-well plates and pretreated with the indicated concentration of fisetin or an equivalent volume of DMSO vehicle control (final concentration of 0.1%) for 30 min, followed by Tm treatment for 16 h.…”

Section: Methodsmentioning

confidence: 99%

Fisetin Protects PC12 Cells from Tunicamycin-Mediated Cell Death via Reactive Oxygen Species Scavenging and Modulation of Nrf2-Driven Gene Expression, SIRT1 and MAPK Signaling in PC12 Cells

Yen

Chen

et al. 2017

IJMS

View full text Add to dashboard Cite

Background: Fisetin (3,7,3′,4′-tetrahydroxyflavone) is a dietary flavonol and exhibits antioxidant, anti-inflammatory, and neuroprotective activities. However, high concentration of fisetin is reported to produce reactive oxygen species (ROS), induce endoplasmic reticulum (ER) stress and cause cytotoxicity in cancer cells. The aim of this study is to investigate the cytoprotective effects of low concentration of fisetin against tunicamycin (Tm)-mediated cytotoxicity in neuronal-like catecholaminergic PC12 cells. Methods: Cell viability was assayed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and apoptotic and autophagic markers were analyzed by Western blot. Gene expression of unfolded protein response (UPR) and Phase II enzymes was further investigated using RT-Q-PCR or Western blotting. Intracellular ROS level was measured using 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA) by a fluorometer. The effects of fisetin on mitogen activated protein kinases (MAPKs) and SIRT1 (Sirtuin 1) signaling pathways were examined using Western blotting and specific inhibitors. Results: Fisetin (<20 µM) restored cell viability and repressed apoptosis, autophagy and ROS production in Tm-treated cells. Fisetin attenuated Tm-mediated expression of ER stress genes, such as glucose-regulated proteins 78 (GRP78), C/EBP homologous protein (CHOP also known as GADD153) and Tribbles homolog 3 (TRB3), but induced the expression of nuclear E2 related factor (Nrf)2-targeted heme oxygenase (HO)-1, glutamate cysteine ligase (GCL) and cystine/glutamate transporter (xCT/SLC7A11), in both the presence and absence of Tm. Moreover, fisetin enhanced phosphorylation of ERK (extracellular signal-regulated kinase), JNK (c-JUN NH2-terminal protein kinase), and p38 MAPK. Addition of JNK and p38 MAPK inhibitor significantly antagonized its cytoprotective activity and modulatory effects on UPR. Fisetin also restored Tm-inhibited SIRT1 expression and addition of sirtinol (SIRT1 activation inhibitor) significantly blocked fisetin-mediated cytoprotection. In conclusion, this result shows that fisetin activates Nrf2, MAPK and SIRT1, which may elicit adaptive cellular stress response pathways so as to protect cells from Tm-induced cytotoxicity.

show abstract

“…It is well known that tunicamycin (Tm) induces ER stress-mediated apoptosis in various cells. We previously reported that the cleavage of caspase-12, which is known as an ER stress-specific caspase [32], is involved in the progression of Tm-mediated apoptosis in cultured PC12 cells [28]. To further elucidate the characteristics of the EDC-induced effects on PC12 cells, we measured the viability of PC12 cells treated with different concentrations of BPA, NP, TBT or beno for 24 hours using the MTT assay (Figure 1).…”

Section: Resultsmentioning

confidence: 99%

“…After cultivation for 24 h, the cells were lysed in lysis buffer as described previously [28]. Total lysate (20 μg per lane) was loaded on SDS-PAGE and blotted onto a PVDF membrane using a semi-dry blotter (Atto, Tokyo, Japan).…”

Section: Immunoblotting Analysismentioning

confidence: 99%

Apoptosis-inducing activity of endocrine-disrupting chemicals in cultured PC12 cells

Sasaya¹,

Yasuzumi²,

Maruoka³

et al. 2012

ABC

View full text Add to dashboard Cite

Endocrine-disrupting chemicals (EDCs) are known to exert estrogen-like effects that are similar to those made by naturally produced hormones or by inhibition of the receptors in the cell receiving the hormones. Recently, several reports have indicated that EDCs can affect the developing central nervous system. In our current study, we report that some EDCs induce apoptosis in cultured PC12 cells and can be classified into three groups. Bisphenol A (BPA), pnonylphenol (NP) and tributyltin chloride (TBT) were found to induce endoplasmic reticulum (ER) stress-associated apoptosis and activate the unfolded protein response (UPR) system, whereas benomyl (beno) induced non-ER stress-associated apoptosis. The half-maximal apoptosis-inducing concentrations (IC 50 ) of these EDCs were 160 µM for BPA, 25.6 µM for NP, 640 nM for TBT and 48 µM for beno. Although these concentrations are higher than those found in the environment, some EDCs may have apoptotic effects on various cells in the body, including neurons, through their accumulation in the body over time or condensation through the food chain. On the other hand, benzopyrene, fenvalerate, styrene monomer and bis(2-ethylhexyl)phthalate did not induce apoptosis in PC12 cells. We analyzed also whether apoptosis-inducing EDCs had an estrogen-like effect on cultured PC12 cells transfected with a luciferase reporter plasmid, the activity of which is dependent on estrogen receptor α. We found that BPA had an estrogen-like effect (EC 50 = 5.9 µM) but that NP, TBT and beno did not in transfected PC12 cells. These results suggest that BPA may predominantly exert estrogenic effects, but others may predominantly have apoptosis-inducing effects on cells in the body exposed to a polluted environment.

show abstract

Nicotine Suppresses Tunicamycin-Induced, But Not Thapsigargin-Induced, Expression of GRP78 during ER Stress-Mediated Apoptosis in PC12 Cells

Cited by 30 publications

References 27 publications

Pharmacological Chaperoning of Nicotinic Acetylcholine Receptors Reduces the Endoplasmic Reticulum Stress Response

Pharmacological Chaperoning of Nicotinic Acetylcholine Receptors Reduces the Endoplasmic Reticulum Stress Response

Fisetin Protects PC12 Cells from Tunicamycin-Mediated Cell Death via Reactive Oxygen Species Scavenging and Modulation of Nrf2-Driven Gene Expression, SIRT1 and MAPK Signaling in PC12 Cells

Apoptosis-inducing activity of endocrine-disrupting chemicals in cultured PC12 cells

Contact Info

Product

Resources

About