RT Journal Article
SR Electronic
T1 Change in Amino Acid Pools During Neuronal Differentiation of PC12 Cells
JF In Vivo
JO In Vivo
FD International Institute of Anticancer Research
SP 1403
OP 1408
DO 10.21873/invivo.11392
VO 32
IS 6
A1 AIKO SANO
A1 HAIXIA SHI
A1 RYUICHIRO SUZUKI
A1 YOSHIAKI SHIRATAKI
A1 HIROSHI SAKAGAMI
YR 2018
UL http://iv.iiarjournals.org/content/32/6/1403.abstract
AB Background/Aim: Although rat PC12 cells are a well-established model to investigate neuronal differentiation, survival and function, the reports of differentiation-associated changes in the intracellular amino acid pools of neurotransmitters have been limited. In this study, possible changes in the intracellular amino acid pools were investigated during nerve growth factor (NGF)-induced differentiation of PC12 cells. Materials and Methods: Rat PC12 cells were induced to differentiate into neuronal cells by 50 ng/ml NGF in serum-free Dulbecco's Modified Eagle's medium, followed by the addition of fresh NGF-containing medium at day 3, without medium change. Cell viability was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Intracellular amino acids were extracted by 5%trichloroacetic acid and quantified by amino acid analyzer. Results: Differentiated PC12 cells showed high concentrations of excitatory neurotransmitters (glutamic acid and aspartic acid) and glutamine (energy supply). In contrast, urea and taurine levels declined with the progression of neuronal differentiation. Exogenous addition of taurine, urea, and L- and D- aspartic acid showed little or no effect on supporting viability of PC12 cells cultured in serum-free medium. Conclusion: The present study demonstrated dramatic changes in the composition of intracellular amino acids during neuronal differentiation.