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首页> 外文期刊>Plant biology >Cytosolic calcium, hydrogen peroxide and related gene expression and protein modulation in Arabidopsis thaliana cell cultures respond immediately to altered gravitation: parabolic flight data
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Cytosolic calcium, hydrogen peroxide and related gene expression and protein modulation in Arabidopsis thaliana cell cultures respond immediately to altered gravitation: parabolic flight data

机译：拟南芥细胞培养物中的胞质钙，过氧化氢及相关基因表达和蛋白质调控对引力变化的响应迅速：抛物线飞行数据

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Callus cell cultures of Arabidopsis thaliana (cv. Columbia) were exposed to parabolic flights in order to assess molecular, short-term responses to altered gravity fields. Using transgenic cell lines, hydrogen peroxide (H_2O_2) and cytosolic Ca~(2+) were continuously monitored. In parallel, the metabolism of samples was chemically quenched (RNAlater, Ambion for RNA; acid/base for NADPH, NADP) at typical stages of a parabola [1 g before pull up; end of pull up (1.8 g), end of microgravity (20 s) and end of pull out (1.8 g)]. Cells exhibited an increase in both Ca~(2+) and H_2O_2 with the onset of microgravity, and a decline thereafter. This behaviour was accompanied by a decrease of the NADPH/NADP redox ratio, indicating Ca~(2+)-dependent activation of a NADPH oxidase. Microarray analyses revealed concomitant expression profiles. At the end of the microgravity phase, 396 transcripts were specifically up-, while 485 were down-regulated. Up-regulation was dominated by Ca~(2+)- and ROS-related gene products. The same material was also used for analysis of phosphopeptides with 2-D SDS PAGE. Relevant spots were identified by liquid chromatography–MS. With the exception of a chaperone (HSP 70-3), hypergravity (1.8 g) and microgravity modified different sets of proteins. These are partly involved in primary metabolism (glycolysis, gluconeogenesis, citrate cycle) and detoxification of ROS. Taken together, these data show that both gene expression and protein modulation jointly respond within seconds to alterations in the gravity field, with a focus on metabolic adaptation, signalling and control of ROS.
机译：将拟南芥（cv。Columbia）的愈伤组织细胞培养物进行抛物线飞行，以评估对重力场变化的分子，短期反应。使用转基因细胞系，连续监测过氧化氢（H_2O_2）和胞质Ca〜（2+）。平行地，在抛物线的典型阶段将样品的代谢化学猝灭（RNAlater，RNA的Ambion; NADPH的酸/碱，NADP）[上拉前1 g;提拉结束（1.8 g），微重力结束（20 s）和提拉结束（1.8 g）]。随着微重力的开始，细胞Ca〜（2+）和H_2O_2均增加，此后下降。该行为伴随着NADPH / NADP氧化还原比的降低，表明NADPH氧化酶的Ca〜（2+）依赖性活化。微阵列分析揭示了伴随的表达谱。在微重力阶段结束时，有396个转录物被特异上调，而485个则被下调。上调主要由Ca〜（2+）和ROS相关基因产物决定。相同的材料也用于2-D SDS PAGE分析磷酸肽。通过液相色谱-MS鉴定相关斑点。除伴侣蛋白（HSP 70-3）外，超重力（1.8 g）和微重力可修饰不同组的蛋白质。它们部分参与ROS的初次代谢（糖酵解，糖异生，柠檬酸盐循环）和解毒。综上所述，这些数据表明基因表达和蛋白质调节在数秒内共同对重力场的变化做出响应，重点是代谢适应，ROS的信号传导和控制。

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