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Genes involved in the astrocyte-neuron lactate shuttle (ANLS) are specifcally regulated in cortical astrocytes following sleep deprivation in mice

机译：小鼠睡眠剥夺后，皮质星形胶质细胞中涉及星形胶质细胞-神经元乳酸穿梭（ANLS）的基因受到特定调节

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Study Objectives: There is growing evidence indicating that in order to meet the neuronal energy demands, astrocytes provide lactate as an energy substrate for neurons through a mechanism called "astrocyte-neuron lactate shuttle" (ANLS). Since neuronal activity changes dramatically during vigilance states, we hypothesized that the ANLS may be regulated during the sleep-wake cycle. To test this hypothesis we investigated the expression of genes associated with the ANLS specifcally in astrocytes following sleep deprivation. Astrocytes were purifed by fuorescence-activated cell sorting from transgenic mice expressing the green fuorescent protein (GFP) under the control of the human astrocytic GFAP-promoter. Design: 6-hour instrumental sleep deprivation (TSD). Setting: Animal sleep research laboratory. Participants: Young (P23-P27) FVB/N-Tg (GFAP-GFP) 14Mes/J (Tg) mice of both sexes and 7-8 week male Tg and FVB/Nj mice. Interventions: Basal sleep recordings and sleep deprivation achieved using a modifed cage where animals were gently forced to move. Measurements and Results: Since Tg and FVB/Nj mice displayed a similar sleep-wake pattern, we performed a TSD in young Tg mice. Total RNA was extracted from the GFP-positive and GFP-negative cells sorted from cerebral cortex. Quantitative RT-PCR analysis showed that levels of Glut1, a-2-Na/K pump, Glt1, and Ldha mRNAs were signifcantly increased following TSD in GFP-positive cells. In GFP-negative cells, a tendency to increase, although not signifcant, was observed for Ldha, Mct2, and α-3-Na/K pump mRNAs. Conclusions: This study shows that TSD induces the expression of genes associated with ANLS specifcally in astrocytes, underlying the important role of astrocytes in the maintenance of the neuro-metabolic coupling across the sleep-wake cycle.
机译：研究目的：越来越多的证据表明，为了满足神经元的能量需求，星形胶质细胞通过称为“星形胶质细胞-神经元乳酸穿梭”（ANLS）的机制提供乳酸作为神经元的能量底物。由于在警惕状态下神经元的活动发生了巨大变化，因此我们假设ANLS可能在睡眠-觉醒周期中受到调节。为了检验该假设，我们研究了睡眠剥夺后星形胶质细胞中与ANLS相关的基因的表达。通过荧光激活细胞分选法从星形胶质细胞GFAP启动子控制下表达绿色荧光蛋白（GFP）的转基因小鼠中纯化星形胶质细胞。设计：6小时仪器性睡眠剥夺（TSD）。地点：动物睡眠研究实验室。参与者：两性的年轻（P23-P27）FVB / N-Tg（GFAP-GFP）14Mes / J（Tg）小鼠和7-8周的雄性Tg和FVB / Nj小鼠。干预措施：使用改良的笼子轻轻地迫使动物移动，从而实现基础睡眠记录和睡眠剥夺。测量和结果：由于Tg和FVB / Nj小鼠表现出相似的睡眠-觉醒模式，因此我们在年轻的Tg小鼠中进行了TSD。从大脑皮层分选的GFP阳性和GFP阴性细胞中提取总RNA。定量RT-PCR分析显示，TSD后GFP阳性细胞中Glut1，a-2-Na / K泵，Glt1和Ldha mRNA的水平显着增加。在GFP阴性细胞中，观察到Ldha，Mct2和α-3-Na/ K泵mRNA表达升高的趋势，尽管并不明显。结论：这项研究表明，TSD特异性地诱导星形胶质细胞中与ANLS相关的基因的表达，这可能是星形胶质细胞在维持整个睡眠-觉醒周期中神经代谢耦合中的重要作用。

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