Aiko Sano1, Haixia Shi2,3, Ryuichiro Suzuki4, Yoshiaki Shirataki1, Hiroshi Sakagami5. 1. Faculty of Pharmacy and Pharmaceutical Sciences, Josai University, Saitama, Japan. 2. Department of Traditional Chinese Medicine, Shanghai Ninth People's Hospital, Shanghai Jiatong University School of Medicine, Shanghai, P.R. China. 3. Meikai University Research Institute of Odontology (M-RIO), Meikai University School of Dentistry, Saitama, Japan. 4. Faculty of Pharmacy and Pharmaceutical Sciences, Josai University, Saitama, Japan ryu_suzu@josai.ac.jp sakagami@dent.meikai.ac.jp. 5. Meikai University Research Institute of Odontology (M-RIO), Meikai University School of Dentistry, Saitama, Japan ryu_suzu@josai.ac.jp sakagami@dent.meikai.ac.jp.
Abstract
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. Copyright
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. Copyright
Authors: Aurélia Ravni; Steve Bourgault; Alexis Lebon; Philippe Chan; Ludovic Galas; Alain Fournier; Hubert Vaudry; Bruno Gonzalez; Lee E Eiden; David Vaudry Journal: J Neurochem Date: 2006-07 Impact factor: 5.372
Authors: Ralph A Bradshaw; Jay Pundavela; Jordane Biarc; Robert J Chalkley; A L Burlingame; Hubert Hondermarck Journal: Adv Biol Regul Date: 2014-11-20