RRC ID |
17629
|
著者 |
Yamada T, Inazu M, Tajima H, Matsumiya T.
|
タイトル |
Functional expression of choline transporter-like protein 1 (CTL1) in human neuroblastoma cells and its link to acetylcholine synthesis.
|
ジャーナル |
Neurochem Int
|
Abstract |
We examined the molecular and functional characterization of choline uptake into human neuroblastoma cell lines (SH-SY5Y: non-cholinergic and LA-N-2: cholinergic neuroblastoma), and the association between choline transport and acetylcholine (ACh) synthesis in these cells. Choline uptake was saturable and mediated by a single transport system. Removal of Na(+) from the uptake buffer strongly enhanced choline uptake. Choline uptake was inhibited by the choline analogue hemicholinium-3 (HC-3) and various organic cations, and was significantly decreased by acidification of the extracellular medium. The increase in choline uptake under Na(+)-free conditions was inhibited by a Na(+)/H(+) exchanger (NHE) inhibitor. Real-time PCR revealed that choline transporter-like protein 1 (CTL1), NHE1 and NHE5 mRNA are mainly expressed. Western blot and immunocytochemical analysis indicated that CTL1 protein was expressed in plasma membrane. ChAT mRNA was expressed at a much higher level in LA-N-2 cells than in SH-SY5Y cells. The conversion of choline to ACh was confirmed in both cells, and was enhanced in Na(+)-free conditions. These findings suggest that CTL1 is functionally expressed in both SH-SY5Y and LA-N-2 cells and is responsible for choline uptake that relies on a directed H(+) gradient as a driving force, and this transport functions in co-operation with NHE1 and NHE5. Furthermore, choline uptake through CTL1 is associated with ACh synthesis in cholinergic neuroblastoma cells.
|
巻・号 |
58(3)
|
ページ |
354-65
|
公開日 |
2011-2-1
|
DOI |
10.1016/j.neuint.2010.12.011
|
PII |
S0197-0186(10)00381-5
|
PMID |
21185344
|
MeSH |
Acetylcholine / biosynthesis*
Animals
Antigens, CD / genetics
Antigens, CD / metabolism*
Brain Neoplasms / metabolism*
COS Cells
Cell Line, Tumor
Chlorocebus aethiops
Humans
Neuroblastoma / metabolism*
Neurons / metabolism*
Organ Culture Techniques
Organic Cation Transport Proteins / genetics
Organic Cation Transport Proteins / metabolism*
Rats
Rats, Wistar
|
IF |
3.881
|
引用数 |
30
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
NEUROSCIENCES
|
リソース情報 |
ヒト・動物細胞 |
LA-N-2(RCB0484) |