Abstract
We have previously demonstrated the functional expression of glutamine (Gln) transporter (GlnT) believed to predominate in neurons for the neurotransmitter glutamate pool by rat neocortical astrocytes devoid of neuronal marker expression, with exacerbated vulnerability to oxidative stress after transient overexpression. To evaluate molecular mechanisms underlying the exacerbation, we established stable GlnT transfectants in rat astrocytic C6 glioma cells. In two different clones of stable transfectants with increased intracellular Gln levels, exposure to hydrogen peroxide (H2O 2) and A23187, but not to tunicamycin or 2,4-dinitrophenol, led to significant exacerbation of the cytotoxicity compared to cells with empty vector (EV). Stable GlnT overexpression led to a significant increase in heme oxygenase-1 protein levels in a manner sensitive to H2O2, whereas H2O2 was significantly more effective in increasing NO2 accumulation and reactive oxygen species (ROS) generation in stable GlnT transfectants than in EV cells. Moreover, exposure to A23187 led to a more effective increase in the generation of ROS in stable GlnT transfectants than in stable EV transfectants. These results suggest that GlnT may play a role in the mechanisms underlying the determination of cellular viability in astrocytes through modulation of intracellular ROS generation.
| Original language | English |
|---|---|
| Pages (from-to) | 504-511 |
| Number of pages | 8 |
| Journal | Neurochemistry International |
| Volume | 58 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - Mar 2011 |
| Externally published | Yes |
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Keywords
- Astrocytes
- Glutamine transporter
- HO-1
- Oxidative stress
- Reactive oxygen species
- slc38a1
ASJC Scopus subject areas
- Cellular and Molecular Neuroscience
- Cell Biology
Cite this
Exacerbated vulnerability to oxidative stress in astrocytic C6 glioma cells with stable overexpression of the glutamine transporter slc38a1. / Ogura, Masato; Takarada, Takeshi; Nakamichi, Noritaka; Kawagoe, Hirofumi; Sako, Aya; Nakazato, Ryota; Yoneda, Yukio.
In: Neurochemistry International, Vol. 58, No. 4, 03.2011, p. 504-511.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Exacerbated vulnerability to oxidative stress in astrocytic C6 glioma cells with stable overexpression of the glutamine transporter slc38a1
AU - Ogura, Masato
AU - Takarada, Takeshi
AU - Nakamichi, Noritaka
AU - Kawagoe, Hirofumi
AU - Sako, Aya
AU - Nakazato, Ryota
AU - Yoneda, Yukio
PY - 2011/3
Y1 - 2011/3
N2 - We have previously demonstrated the functional expression of glutamine (Gln) transporter (GlnT) believed to predominate in neurons for the neurotransmitter glutamate pool by rat neocortical astrocytes devoid of neuronal marker expression, with exacerbated vulnerability to oxidative stress after transient overexpression. To evaluate molecular mechanisms underlying the exacerbation, we established stable GlnT transfectants in rat astrocytic C6 glioma cells. In two different clones of stable transfectants with increased intracellular Gln levels, exposure to hydrogen peroxide (H2O 2) and A23187, but not to tunicamycin or 2,4-dinitrophenol, led to significant exacerbation of the cytotoxicity compared to cells with empty vector (EV). Stable GlnT overexpression led to a significant increase in heme oxygenase-1 protein levels in a manner sensitive to H2O2, whereas H2O2 was significantly more effective in increasing NO2 accumulation and reactive oxygen species (ROS) generation in stable GlnT transfectants than in EV cells. Moreover, exposure to A23187 led to a more effective increase in the generation of ROS in stable GlnT transfectants than in stable EV transfectants. These results suggest that GlnT may play a role in the mechanisms underlying the determination of cellular viability in astrocytes through modulation of intracellular ROS generation.
AB - We have previously demonstrated the functional expression of glutamine (Gln) transporter (GlnT) believed to predominate in neurons for the neurotransmitter glutamate pool by rat neocortical astrocytes devoid of neuronal marker expression, with exacerbated vulnerability to oxidative stress after transient overexpression. To evaluate molecular mechanisms underlying the exacerbation, we established stable GlnT transfectants in rat astrocytic C6 glioma cells. In two different clones of stable transfectants with increased intracellular Gln levels, exposure to hydrogen peroxide (H2O 2) and A23187, but not to tunicamycin or 2,4-dinitrophenol, led to significant exacerbation of the cytotoxicity compared to cells with empty vector (EV). Stable GlnT overexpression led to a significant increase in heme oxygenase-1 protein levels in a manner sensitive to H2O2, whereas H2O2 was significantly more effective in increasing NO2 accumulation and reactive oxygen species (ROS) generation in stable GlnT transfectants than in EV cells. Moreover, exposure to A23187 led to a more effective increase in the generation of ROS in stable GlnT transfectants than in stable EV transfectants. These results suggest that GlnT may play a role in the mechanisms underlying the determination of cellular viability in astrocytes through modulation of intracellular ROS generation.
KW - Astrocytes
KW - Glutamine transporter
KW - HO-1
KW - Oxidative stress
KW - Reactive oxygen species
KW - slc38a1
UR - http://www.scopus.com/inward/record.url?scp=79952104569&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79952104569&partnerID=8YFLogxK
U2 - 10.1016/j.neuint.2011.01.007
DO - 10.1016/j.neuint.2011.01.007
M3 - Article
C2 - 21219957
AN - SCOPUS:79952104569
VL - 58
SP - 504
EP - 511
JO - Neurochemistry International
JF - Neurochemistry International
SN - 0197-0186
IS - 4
ER -