Trophos, Parc Scientifique de Luminy, Luminy Biotech Entreprises, Marseille, France.
PLoS One. 2013 Nov 26;8(11):e81528. doi: 10.1371/journal.pone.0081528. eCollection 2013.
In Huntington disease (HD), there is increasing evidence for a link between mutant huntingtin expression, mitochondrial dysfunction, energetic deficits and neurodegeneration but the precise nature, causes and order of these events remain to be determined. In this work, our objective was to evaluate mitochondrial respiratory function in intact, non-permeabilized, neurons derived from a transgenic rat model for HD compared to their wild type littermates by measuring oxygen consumption rates and extracellular acidification rates. Although HD striatal neurons had similar respiratory capacity as those from their wild-type littermates when they were incubated in rich medium containing a supra-physiological glucose concentration (25 mM), pyruvate and amino acids, respiratory defects emerged when cells were incubated in media containing only a physiological cerebral level of glucose (2.5 mM). According to the concept that glucose is not the sole substrate used by the brain for neuronal energy production, we provide evidence that primary neurons can use lactate as well as pyruvate to fuel the mitochondrial respiratory chain. In contrast to glucose, we found no major deficits in HD striatal neurons' capacity to use pyruvate as a respiratory substrate compared to wild type littermates. Additionally, we used extracellular acidification rates to confirm a reduction in anaerobic glycolysis in the same cells. Interestingly, the metabolic disturbances observed in striatal neurons were not seen in primary cortical neurons, a brain region affected in later stages of HD. In conclusion, our results argue for a dysfunction in glycolysis, which might precede any defects in the respiratory chain itself, and these are early events in the onset of disease.
在亨廷顿病(HD)中,越来越多的证据表明突变亨廷顿蛋白的表达、线粒体功能障碍、能量不足和神经退行性变之间存在联系,但这些事件的确切性质、原因和顺序仍有待确定。在这项工作中,我们的目的是通过测量耗氧量和细胞外酸化率来评估来自 HD 转基因大鼠模型的未通透、完整神经元与野生型同窝仔鼠的线粒体呼吸功能。尽管 HD 纹状体神经元在富含含有超生理葡萄糖浓度(25 mM)、丙酮酸和氨基酸的培养基中孵育时具有与野生型同窝仔鼠相似的呼吸能力,但当细胞在仅含有生理脑葡萄糖水平(2.5 mM)的培养基中孵育时,呼吸缺陷就会出现。根据葡萄糖不是大脑用于神经元能量产生的唯一底物的概念,我们提供了证据表明,原代神经元可以使用乳酸盐以及丙酮酸来为线粒体呼吸链供能。与葡萄糖不同,我们发现与野生型同窝仔鼠相比,HD 纹状体神经元利用丙酮酸作为呼吸底物的能力没有重大缺陷。此外,我们使用细胞外酸化率来证实同一细胞中无氧糖酵解的减少。有趣的是,在纹状体神经元中观察到的代谢紊乱在初级皮质神经元中没有出现,而初级皮质神经元是 HD 晚期受影响的大脑区域。总之,我们的结果表明糖酵解功能障碍,这可能先于呼吸链本身的任何缺陷,并且这些是疾病发作的早期事件。