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)