Abstract
The purpose of the present study was to elucidate the expression and regulation of the L-cystine transporter, system xc -, in Müller cells. In this study, newly developed conditionally immortalized rat Müller cell lines (TR-MUL) from transgenic rats harboring the temperature-sensitive SV 40 large T-antigen gene were used as an in vitro model. TR-MUL cells express large T-antigen and grow well at 33°C with a doubling time of 30 h, but do not grow at 39°C. TR-MUL cells express typical Müller cell markers such as S-100, glutamine synthetase, and EAAT1/GLAST, whereas EAAT2/GLT-1 and EAAT5 are not detected. TR-MUL cells also exhibit little or no expression of glial fibrillary acidic protein. We found that TR-MUL5 cells exhibited [14C]L-cystine uptake activity and expressed xCT and 4F2hc, which involve system xc -. The uptake of [14C]L-cystine was significantly inhibited by L-glutamic acid and L-aspartic acid, whereas L-leucine had no effect. Following diethyl maleate (DEM) treatment, the glutathione concentration in TR-MUL5 cells was reduced in the first 24 h, then gradually recovered for more than 24 h. The L-cystine uptake rate and the xCT expression level in TR-MUL5 cells were enhanced by DEM treatment. In contrast, the 4F2hc expression level was unchanged. In conclusion, TR-MUL cells have the properties of Müller cells and exhibit system x c --mediated L-cystine uptake activity. The oxidative stress conditions following DEM treatment activate L-cystine transport in TR-MUL cells due to the enhanced transcription of the xCT gene.
| Original language | English |
|---|---|
| Pages (from-to) | 208-217 |
| Number of pages | 10 |
| Journal | GLIA |
| Volume | 43 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - 2003 Sep 1 |
| Externally published | Yes |
Fingerprint
Keywords
- Conditionally immortalized cell line
- Diethyl maleate
- Glutathione
- Oxidative stress
ASJC Scopus subject areas
- Immunology
Cite this
Expression and regulation of L-cystine transporter, system X c -, in the newly developed rat retinal Müller cell line (TR-MUL). / Tomi, Masatoshi; Funaki, Takeshi; Abukawa, Hayato; Katayama, Kazunori; Kondo, Tetsu; Ohtsuki, Sumio; Ueda, Masatsugu; Obinata, Masuo; Terasaki, Tetsuya; Hosoya, Ken Ichi.
In: GLIA, Vol. 43, No. 3, 01.09.2003, p. 208-217.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Expression and regulation of L-cystine transporter, system X c -, in the newly developed rat retinal Müller cell line (TR-MUL)
AU - Tomi, Masatoshi
AU - Funaki, Takeshi
AU - Abukawa, Hayato
AU - Katayama, Kazunori
AU - Kondo, Tetsu
AU - Ohtsuki, Sumio
AU - Ueda, Masatsugu
AU - Obinata, Masuo
AU - Terasaki, Tetsuya
AU - Hosoya, Ken Ichi
PY - 2003/9/1
Y1 - 2003/9/1
N2 - The purpose of the present study was to elucidate the expression and regulation of the L-cystine transporter, system xc -, in Müller cells. In this study, newly developed conditionally immortalized rat Müller cell lines (TR-MUL) from transgenic rats harboring the temperature-sensitive SV 40 large T-antigen gene were used as an in vitro model. TR-MUL cells express large T-antigen and grow well at 33°C with a doubling time of 30 h, but do not grow at 39°C. TR-MUL cells express typical Müller cell markers such as S-100, glutamine synthetase, and EAAT1/GLAST, whereas EAAT2/GLT-1 and EAAT5 are not detected. TR-MUL cells also exhibit little or no expression of glial fibrillary acidic protein. We found that TR-MUL5 cells exhibited [14C]L-cystine uptake activity and expressed xCT and 4F2hc, which involve system xc -. The uptake of [14C]L-cystine was significantly inhibited by L-glutamic acid and L-aspartic acid, whereas L-leucine had no effect. Following diethyl maleate (DEM) treatment, the glutathione concentration in TR-MUL5 cells was reduced in the first 24 h, then gradually recovered for more than 24 h. The L-cystine uptake rate and the xCT expression level in TR-MUL5 cells were enhanced by DEM treatment. In contrast, the 4F2hc expression level was unchanged. In conclusion, TR-MUL cells have the properties of Müller cells and exhibit system x c --mediated L-cystine uptake activity. The oxidative stress conditions following DEM treatment activate L-cystine transport in TR-MUL cells due to the enhanced transcription of the xCT gene.
AB - The purpose of the present study was to elucidate the expression and regulation of the L-cystine transporter, system xc -, in Müller cells. In this study, newly developed conditionally immortalized rat Müller cell lines (TR-MUL) from transgenic rats harboring the temperature-sensitive SV 40 large T-antigen gene were used as an in vitro model. TR-MUL cells express large T-antigen and grow well at 33°C with a doubling time of 30 h, but do not grow at 39°C. TR-MUL cells express typical Müller cell markers such as S-100, glutamine synthetase, and EAAT1/GLAST, whereas EAAT2/GLT-1 and EAAT5 are not detected. TR-MUL cells also exhibit little or no expression of glial fibrillary acidic protein. We found that TR-MUL5 cells exhibited [14C]L-cystine uptake activity and expressed xCT and 4F2hc, which involve system xc -. The uptake of [14C]L-cystine was significantly inhibited by L-glutamic acid and L-aspartic acid, whereas L-leucine had no effect. Following diethyl maleate (DEM) treatment, the glutathione concentration in TR-MUL5 cells was reduced in the first 24 h, then gradually recovered for more than 24 h. The L-cystine uptake rate and the xCT expression level in TR-MUL5 cells were enhanced by DEM treatment. In contrast, the 4F2hc expression level was unchanged. In conclusion, TR-MUL cells have the properties of Müller cells and exhibit system x c --mediated L-cystine uptake activity. The oxidative stress conditions following DEM treatment activate L-cystine transport in TR-MUL cells due to the enhanced transcription of the xCT gene.
KW - Conditionally immortalized cell line
KW - Diethyl maleate
KW - Glutathione
KW - Oxidative stress
UR - http://www.scopus.com/inward/record.url?scp=0042866085&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0042866085&partnerID=8YFLogxK
U2 - 10.1002/glia.10253
DO - 10.1002/glia.10253
M3 - Article
C2 - 12898700
AN - SCOPUS:0042866085
VL - 43
SP - 208
EP - 217
JO - GLIA
JF - GLIA
SN - 0894-1491
IS - 3
ER -