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Kinetic properties of the redox switch/redox coupling mechanism as determined in primary cultures of cortical neurons and astrocytes from rat brain

机译：在大鼠大脑皮质神经元和星形胶质细胞的原代培养中确定的氧化还原开关/氧化还原耦合机制的动力学性质

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We investigate the mechanisms underlying the redox switch/redox coupling hypothesis by characterizing the competitive consumption of glucose or lactate and the kinetics of pyruvate production in primary cultures of cortical neurons and astrocytes from rat brain. Glucose consumption was determined in neuronal cultures incubated in Krebs ringer bicarbonate buffer (KRB) containing 0.25–5 mM glucose, in the presence and absence of 5 mM lactate as an alternative substrate. Lactate consumption was measured in neuronal cultures incubated with 1–15 mM lactate, in the presence and absence of 1 mM glucose. In both cases, the alternative substrate increased the Km (mM) values for glucose consumption (from 2.2 ± 0.2 to 3.6 ± 0.1) or lactate consumption (from 7.8 ± 0.1 to 8.5 ± 0.1) without significant changes on the corresponding Vmax. This is consistent with a competitive inhibition between the simultaneous consumption of glucose and lactate. When cultures of neurons or astrocytes were incubated with increasing lactate concentrations 1–20 mM, pyruvate production was observed with Km (mM) and Vmax (nmol/mg/h) values of 1.0 ± 0.1 and 109 ± 4 in neurons, or 0.28 ± 0.1 and 342 ± 54 in astrocytes. Thus, astrocytes or neurons are able to return to the incubation medium as pyruvate, a significant part of the lactate consumed. Present results support the reversible exchange of reducing equivalents between neurons and astrocytes in the form of lactate or pyruvate. Monocarboxylate exchange is envisioned to operate under near equilibrium, with the transcellular flux directed thermodynamically toward the more oxidized intracellular redox environment.
机译：我们通过表征葡萄糖或乳酸的竞争性消耗以及大鼠大脑皮层神经元和星形胶质细胞的原代培养物中丙酮酸产生的动力学来研究氧化还原开关/氧化还原耦合假说的潜在机制。在存在和不存在5 mM乳酸作为替代底物的情况下，在含有0.25-5 mM葡萄糖的克雷布斯林格氏碳酸氢盐缓冲液（KRB）中孵育的神经元培养物中测定葡萄糖消耗。在有或没有1 mM葡萄糖的情况下，在与1–15 mM乳酸孵育的神经元培养物中测量乳酸的消耗。在这两种情况下，替代底物均使葡萄糖消耗（从2.2±0.2到3.6±0.1）或乳酸消耗（从7.8±0.1到8.5±0.1）的Km（mM）值增加，而相应的Vmax却没有明显变化。这与葡萄糖和乳酸的同时消耗之间的竞争性抑制相一致。将神经元或星形胶质细胞培养物与增加的乳酸浓度1–20 mM一起孵育时，观察到丙酮酸的产生，神经元的Km（mM）和Vmax（nmol / mg / h）值为1.0±0.1和109±4，或0.28±在星形胶质细胞中为0.1和342±54。因此，星形胶质细胞或神经元能够以丙酮酸（消耗的乳酸的很大一部分）的形式返回培养液。目前的结果支持乳酸或丙酮酸形式的神经元和星形胶质细胞之间还原性当量的可逆交换。设想单羧酸盐交换在接近平衡下进行，其中跨细胞通量以热力学方式指向更氧化的细胞内氧化还原环境。

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