PC12细胞神经元分化过程中氨基酸池的变化
Change in Amino Acid Pools During Neuronal Differentiation of PC12 Cells.
作者信息
Sano Aiko, Shi Haixia, Suzuki Ryuichiro, Shirataki Yoshiaki, Sakagami Hiroshi
机构信息
Faculty of Pharmacy and Pharmaceutical Sciences, Josai University, Saitama, Japan.
Department of Traditional Chinese Medicine, Shanghai Ninth People's Hospital, Shanghai Jiatong University School of Medicine, Shanghai, P.R. China.
出版信息
In Vivo. 2018 Nov-Dec;32(6):1403-1408. doi: 10.21873/invivo.11392.
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.
背景/目的：尽管大鼠嗜铬细胞瘤（PC12）细胞是研究神经元分化、存活及功能的成熟模型，但有关神经递质细胞内氨基酸池分化相关变化的报道却很有限。本研究调查了在神经生长因子（NGF）诱导PC12细胞分化过程中细胞内氨基酸池可能发生的变化。
材料与方法
在无血清的杜氏改良 Eagle 培养基中，用 50 ng/ml NGF 诱导大鼠 PC12 细胞分化为神经元细胞，于第 3 天添加新鲜的含 NGF 培养基，不更换培养基。采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法测定细胞活力。用 5%三氯乙酸提取细胞内氨基酸，并用氨基酸分析仪进行定量分析。
结果
分化的 PC12 细胞显示出高浓度的兴奋性神经递质（谷氨酸和天冬氨酸）和谷氨酰胺（能量供应）。相比之下，尿素和牛磺酸水平随着神经元分化进程而下降。外源添加牛磺酸、尿素以及 L-和 D-天冬氨酸对支持无血清培养基中培养的 PC12 细胞活力几乎没有影响。
结论
本研究证明了神经元分化过程中细胞内氨基酸组成发生了显著变化。
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