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首页> 外文会议>Proceedings of Life in space for life on earth >JOINT EUROPEAN PARTIAL-G PARABOLIC FLIGHT CAMPAIGN CALCIUM ANALYSIS IN ARABIDOPSIS THALIANA CELL CULTURES
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JOINT EUROPEAN PARTIAL-G PARABOLIC FLIGHT CAMPAIGN CALCIUM ANALYSIS IN ARABIDOPSIS THALIANA CELL CULTURES

机译：阿拉伯拟南芥细胞培养物中的联合欧洲Partial-G抛物线飞行运动钙分析

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Callus cells derived from stem tissue suspensionrncultures of Arabidopsis thaliana (cv. Columbia) werernexposed to parabolic flights on board of an AirbusrnA300 (Novespace). The cells were either wild type orrnexpressed a fluorescent probe for the quantification ofrncytosolic calcium (green fluorescent protein (GFP)-rnbased Cameleon). The wild type cells were used forrnboth, fluorescence background control, and the analysisrnof gene expression.rnWith respect to fluorescence measurements, changes inrnthe amounts of Ca~(2+), an important component ofrnsignalling chains, could be assayed in vivo in real time.rnThe technique used takes advantage of a shift inrnfluorescence from 480 to 535 nm with increasing Ca~(2+)rncontent.rnDuring the experiment, fluorescence data werernmonitored at Mars, Moon and micro gravity producingrnflight profiles, each at 1g, pull up (1.8g), about 20 to 26rns of mars (0.36g), moon (0.16g) or micro gravity, andrnpull out (1.8g) for 12/12/6 consecutive parabolas atrndifferent days.rnTransition from hypergravity to microgravity resultedrnin a typical increase in cytosolic Ca~(2+). The flight profilern“Moon” (0.16g) exhibited a very similar behaviour asrnmicrogravity, whereas simulation of “Mars” gravitationrn(0.36) resulted in a weaker signal. This can also berndeduced from minimal/maximal values of the ratiornbetween hyper-g and onset of reduced g. Obviously,rnthe threshold gravitation for a Ca~(2+) response is abovern0.36g. Increasing gravity by centrifugation, in contrast,rninduced a decrease in cytosolic calcium. Here, arnthreshold in response was obvious between 3 and 4g.rnIn order to assay changes in gene expression, wernadditionally quenched parabolic flight samples by therninjection of RNAlater. A microarray analysis of thesernsamples showed a clear impact of the different profiles.rnBoth Moon and Mars profiles exhibited less responsernthan the μg profile. However, the latter responded alsornless compared to previous “μg only” flights. In thosernwe had much higher numbers of altered transcripts. Forrnexample, after 20 s of μg we found about 400rntranscripts increased and about 500 transcriptsrndecreased in amount; this compares to 140 /100 only inrnthe present example. We thus assume that thernperturbations induced by Moon and Mars parabolasrndecrease the sensitivity towards μg. We can, however,rndirectly compare the Moon profile with our “μg only”rnexperiments, because these were the first ones withinrnthe whole series to experience reduced gravitation.rnHere, we find only about 20 transcripts up- and aboutrn50 transcripts down-regulated, which is much less thanrnobserved for μg.rnIn summary, we find that Ca~(2+) responses have arnthreshold above Mars gravitation (0.38 g), whilernprominent changes in gene expression occur belowrnMoon gravitation (0.16 g).
机译：将来自拟南芥（cv。Columbia）的干组织悬浮培养物的愈伤组织细胞暴露于AirbusrnA300（Novespace）的抛物线飞行中。细胞可以是野生型的，也可以是荧光表达的荧光探针，用于定量胞质钙（基于绿色荧光蛋白（GFP）的Cameleon）。将野生型细胞用于荧光背景控制和分析基因表达。在荧光测量方面，可以实时检测体内信号传递链的重要组成部分Ca〜（2+）量的变化。所使用的技术利用随Ca〜（2+）含量增加而从480 nm转变为535 nm的荧光。rn在实验过程中，对火星，月球和微重力产生的荧光数据进行了监测，每种飞行谱为1 g，上拉（1.8 g ），约20到26毫秒的火星（0.36克），月球（0.16克）或微重力，然后在连续的12/12/6天连续拉出（1.8克）抛物线。从超重力到微重力的过渡导致胞浆的典型增加Ca〜（2+）。飞行轮廓“月球”（0.16g）表现出非常相似的微重力行为，而“火星”重力（0.36）的模拟导致信号较弱。这也可以从hyper-g与减少的g的发作之间的比率的最小/最大值得出。显然，Ca〜（2+）响应的门槛引力大于0.36g。相反，通过离心增加重力会导致胞质钙的减少。在这里，在3g和4g之间反应的阈值很明显。为了测定基因表达的变化，通过注射RNAlater来额外淬灭抛物线飞行样品。对样品的微阵列分析显示了不同轮廓的明显影响。月亮和火星轮廓均表现出比μg轮廓小的响应。但是，与之前的“仅μg”飞行相比，后者的反应也丝毫不差。在那些书中，我们有更多的抄写本。例如，在20 s微克后，我们发现约400个转录物增加，约500个转录物减少。相比之下，本示例仅为140/100。因此，我们假设由月亮和火星的抛物线引起的摄动降低了对微克的敏感性。但是，我们可以将月球轮廓与我们的“仅μg”实验直接进行比较，因为这是整个系列中第一个经历引力降低的实验。在这里，我们发现仅约20个转录本上调而约50个转录本下调，总之，我们发现Ca〜（2+）反应在火星引力（0.38 g）以上具有一定的阈值，而基因表达的显着变化发生在月球引力（0.16 g）以下。

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  • 会议地点 Aberdeen(GB)
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    Maren Neef; Svenja Fengler; Margret Ecke; Niklas Hausmann; Rüdiger Hampp;

  • 作者单位

    University of Tübingen, Physiological Ecology of Plants, Auf der Morgenstelle 1, 72116 Tübingen (Germany), Email: maren.neef@uni-tuebingen.de;

    University of Tübingen, Physiological Ecology of Plants, Auf der Morgenstelle 1, 72116 Tübingen (Germany);

    University of Tübingen, Physiological Ecology of Plants, Auf der Morgenstelle 1, 72116 Tübingen (Germany);

    University of Tübingen, Physiological Ecology of Plants, Auf der Morgenstelle 1, 72116 Tübingen (Germany);

    University of Tübingen, Physiological Ecology of Plants, Auf der Morgenstelle 1, 72116 Tübingen (Germany), Email: ruediger.hampp@uni-tuebingen.de;

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