Different preferences in the utilization of amino acids for glutathione synthesis in cultured neurons and astroglial cells derived from rat brain
Different preferences in the utilization of amino acids for glutathione synthesis in cultured neurons and astroglial cells derived from rat brain
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DOI：
10.1016/s0304-3940(96)13217-1
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发表时间：
1996-11-29
影响因子：
2.5
通讯作者：
Dringen, R
中科院分区：
文献类型：
--
作者：
Kranich, O;Hamprecht, B;Dringen, R
从胚胎和新生大鼠脑中获得的富含神经元和富含星形胶质细胞的原代培养物中，谷胱甘肽的细胞内含量分别为23.1+/-3.0和31.2+/-6.5 nmol/mg蛋白质。氨基酸脱氨4小时减少了24%，在神经元丰富的文化谷胱甘肽的水平。在再喂食半胱氨酸、谷氨酰胺和甘氨酸时再合成谷氨酰胺。谷胱甘肽的最大含量被发现4小时后再喂养，超过未经处理的神经元丰富的文化的84%。胱氨酸或谷氨酰胺的谷氨酸在4小时的再喂养期间的半胱氨酸的替代导致细胞内谷胱甘肽的量较低。发现如果培养基中补充有250 μ M半胱氨酸，则富含神经元的培养物的谷胱甘肽水平增加76%。然而，如果使用胱氨酸代替，则不会发生这种增加。相反，神经元丰富的文化，星形胶质细胞丰富的原代培养物恢复了最大含量的谷胱甘肽，如果谷氨酸和胱氨酸后，氨基酸剥夺。这些结果表明，半胱氨酸是限制性化合物在培养基中的谷胱甘肽合成的神经元丰富的文化和星形胶质细胞和神经元在文化中有不同的偏好摄取和利用的氨基酸谷胱甘肽合成。
The intracellular contents of glutathione in neuron-rich and astroglia-rich primary cultures derived from the brains of embryonal and newborn rats were found to be 23.1+/-3.0 and 31.2+/-6.5 nmol/mg of protein, respectively. Deprivation of amino acids for 4 h reduced the level of glutathione in neuron-rich cultures by 24%. Glutathione was resynthesized on refeeding of cysteine, glutamine, and glycine. A maximal content of glutathione was found 4 h after refeeding, exceeding that of untreated neuron-rich cultures by 84%. Replacement of cysteine by cystine or glutamine by glutamate during the 4 h refeeding period resulted in a lower intracellular amount of glutathione. An increase in the glutathione level of neuron-rich cultures by 76% was found if the culture medium was supplemented with 250 mu M cysteine. However, no such increase occurred if cystine was used instead. In contrast to neuron-rich cultures, astroglia-rich primary cultures restored a maximal content of glutathione if glutamate and cystine were refed after amino acid deprivation. These results demonstrate that cysteine is the limiting compound in the culture medium for glutathione synthesis in neuron-rich cultures and that astroglial cells and neurons in culture have different preferences for uptake and utilization of amino acids for glutathione synthesis.