Mason Graeme F, Petersen Kitt Falk, de Graaf Robin A, Shulman Gerald I, Rothman Douglas L
Department of Psychiatry, Yale University, School of Medicine, New Haven, Connecticut 06520-8043, USA.
J Neurochem. 2007 Jan;100(1):73-86. doi: 10.1111/j.1471-4159.2006.04200.x. Epub 2006 Oct 31.
Recent studies in rodent and human cerebral cortex have shown that glutamate-glutamine neurotransmitter cycling is rapid and the major pathway of neuronal glutamate repletion. The rate of the cycle remains controversial in humans, because glutamine may come either from cycling or from anaplerosis via glial pyruvate carboxylase. Most studies have determined cycling from isotopic labeling of glutamine and glutamate using a [1-(13)C]glucose tracer, which provides label through neuronal and glial pyruvate dehydrogenase or via glial pyruvate carboxylase. To measure the anaplerotic contribution, we measured (13)C incorporation into glutamate and glutamine in the occipital-parietal region of awake humans while infusing [2-(13)C]glucose, which labels the C2 and C3 positions of glutamine and glutamate exclusively via pyruvate carboxylase. Relative to [1-(13)C]glucose, [2-(13)C]glucose provided little label to C2 and C3 glutamine and glutamate. Metabolic modeling of the labeling data indicated that pyruvate carboxylase accounts for 6 +/- 4% of the rate of glutamine synthesis, or 0.02 micromol/g/min. Comparison with estimates of human brain glutamine efflux suggests that the majority of the pyruvate carboxylase flux is used for replacing glutamate lost due to glial oxidation and therefore can be considered to support neurotransmitter trafficking. These results are consistent with observations made with arterial-venous differences and radiotracer methods.
近期对啮齿动物和人类大脑皮层的研究表明,谷氨酸 - 谷氨酰胺神经递质循环迅速,是神经元谷氨酸补充的主要途径。在人类中,该循环速率仍存在争议,因为谷氨酰胺可能来自循环或通过胶质细胞丙酮酸羧化酶的回补反应。大多数研究使用[1 - (13)C]葡萄糖示踪剂通过谷氨酰胺和谷氨酸的同位素标记来确定循环,该示踪剂通过神经元和胶质细胞丙酮酸脱氢酶或通过胶质细胞丙酮酸羧化酶提供标记。为了测量回补反应的贡献,我们在输注[2 - (13)C]葡萄糖时测量了清醒人类枕顶叶区域中(13)C掺入谷氨酸和谷氨酰胺的情况,[2 - (13)C]葡萄糖仅通过丙酮酸羧化酶标记谷氨酰胺和谷氨酸的C2和C3位置。相对于[1 - (13)C]葡萄糖,[2 - (13)C]葡萄糖为C2和C3谷氨酰胺及谷氨酸提供的标记很少。对标记数据的代谢建模表明,丙酮酸羧化酶占谷氨酰胺合成速率的6±4%,即0.02微摩尔/克/分钟。与人类脑谷氨酰胺流出量的估计值比较表明,丙酮酸羧化酶通量的大部分用于补充因胶质细胞氧化而损失的谷氨酸,因此可认为是支持神经递质运输的。这些结果与通过动静脉差异和放射性示踪剂方法所做的观察结果一致。