Changes in intracellular copper concentration and copper-regulating gene expression after PC12 differentiation into neurons.
It is suspected that some neurodegenerative diseases are a result of the disturbance of copper (Cu) homeostasis, although it remains unclear whether the disturbance of Cu homeostasis has aberrant effects on neurons. Herein, we investigated Cu metabolism specifically in neurons in terms of changes in the intracellular Cu concentration and the expression of Cu-regulating genes, such as Cu transporters and metallothioneins (MTs), before and after the differentiation of rat pheochromocytoma cells (PC12 cells) into neurons. After the differentiation, Cu and Zn imaging with fluorescent probes revealed an increase in intracellular Cu concentration. The concentrations of other essential metals, which were determined by an inductively coupled plasma mass spectrometer, were not altered. The mRNA expression of the Cu influx transporter, Ctr1, was decreased after the differentiation, and the differentiated cells acquired tolerance to Cu and cisplatin, another substrate of Ctr1. In addition, the expression of MT-3, a brain-specific isoform, was increased, contrary to the decreased expression of MT-1 and MT-2. Taken together, the differentiation of PC12 cells into neurons induced MT-3 expression, thereby resulting in intracellular Cu accumulation. The decrease in Ctr1 expression was assumed to be a response aimed at abolishing the physiological accumulation of Cu after the differentiation.
Laboratory of Toxicology and Environmental Health, Graduate School of Pharmaceutical Sciences, Chiba University, Chuo, Chiba 260-8675, Japan.|Laboratory of Chemical Toxicology and Environmental Health, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan.
Yasumitsu
Y
Ogra
Laboratory of Chemical Toxicology and Environmental Health, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan.
Aya
A
Tejima
Laboratory of Chemical Toxicology and Environmental Health, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan.
Naohiro
N
Hatakeyama
Laboratory of Chemical Toxicology and Environmental Health, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan.
Moeko
M
Shiraiwa
Department of Medicinal Organic Chemistry, Graduate School of Pharmaceutical Sciences, Chiba University, Chuo, Chiba 260-8675, Japan.
Siyuan
S
Wu
Department of Medicinal Organic Chemistry, Graduate School of Pharmaceutical Sciences, Chiba University, Chuo, Chiba 260-8675, Japan.
Tsutomu
T
Ishikawa
Laboratory of Chemical Toxicology and Environmental Health, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan.
Ayako
A
Yawata
Laboratory of Chemical Toxicology and Environmental Health, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan.
Yasumi
Y
Anan
Laboratory of Toxicology and Environmental Health, Graduate School of Pharmaceutical Sciences, Chiba University, Chuo, Chiba 260-8675, Japan.|Department of Medicinal Organic Chemistry, Graduate School of Pharmaceutical Sciences, Chiba University, Chuo, Chiba 260-8675, Japan.
Noriyuki
N
Suzuki
D000818:Animals; D027682:Cation Transport Proteins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D003300:Copper / Q000378:metabolism*; D000079923:Copper Transporter 1; D015870:Gene Expression; D006706:Homeostasis; D008668:Metallothionein / Q000235:genetics / Q000378:metabolism; D000090023:Metallothionein 3; D008670:Metals / Q000633:toxicity; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D016716:PC12 Cells; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D015032:Zinc / Q000378:metabolism
D027682:Cation Transport Proteins; D000079923:Copper Transporter 1; D000090023:Metallothionein 3; D008670:Metals; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; C510533:Slc31a1 protein, rat; D003300:Copper; D008668:Metallothionein; D015032:Zinc