Richter P, Lebert M, Tahedl H, Hader D P
Institut fur Botanik und Pharmazeutische Biologie, Friedrich-Alexander-Universitat, Erlangen, Germany.
J Plant Physiol. 2001 Jun;158(6):689-97. doi: 10.1078/0176-1617-00002.
The colorless flagellate Astasia longa shows a pronounced negative gravitaxis. The calcium fluorescence indicator Calcium Crimson was used to detect changes of the intracellular calcium concentration during gravitactical orientation. Astasia shows an increase of the fluorescence after a lag phase of about 10 s, a maximum after about 30 s and a decrease to the basic level within 60 s during gravitactic reorientation. The observed change in fluorescence corresponds to an almost doubling of the initial free calcium concentration. The influence of inhibitors, known to impair gravitaxis, on the calcium concentration of Astasia longa was tested. Addition of caffeine, an inhibitor of phosphodiesterase, increases, while addition of gadolinium, an inhibitor of mechanosensitive ion channels decreases the fluorescence signal. While gravitactic stimulation of caffeine-treated cells resulted in a kinetics of fluorescence intensity changes comparable to control cells the addition of gadolinium inhibited any calcium concentration change. Dynamic fluorescence imaging was used during a sounding rocket experiment (MAXUS 3 campaign). Different accelerations interrupted by microgravity intervals were applied to Astasia cells. The cells show an increase in the calcium signal upon acceleration and a decrease during the microgravity state. The results strongly reemphasize the working model of gravitaxis which is based on the activation of mechano-sensitive ion channels as one of the primary events in signal perception.
无色鞭毛虫长眼虫表现出明显的负重力趋性。使用钙荧光指示剂钙 Crimson 来检测重力定向过程中细胞内钙浓度的变化。长眼虫在重力重新定向过程中,经过约10秒的延迟期后荧光增强,约30秒后达到最大值,60秒内降至基础水平。观察到的荧光变化对应于初始游离钙浓度几乎翻倍。测试了已知会损害重力趋性的抑制剂对长眼虫钙浓度的影响。添加磷酸二酯酶抑制剂咖啡因会使荧光信号增强,而添加机械敏感离子通道抑制剂钆会降低荧光信号。虽然对咖啡因处理的细胞进行重力刺激导致荧光强度变化的动力学与对照细胞相当,但添加钆会抑制任何钙浓度变化。在一次探空火箭实验(MAXUS 3任务)中使用了动态荧光成像。对长眼虫细胞施加了由微重力间隔中断的不同加速度。细胞在加速时钙信号增强,在微重力状态下减弱。这些结果有力地再次强调了基于机械敏感离子通道激活作为信号感知主要事件之一的重力趋性工作模型。