Bulychev Alexander A, Komarova Anna V
Faculty of Biology, Lomonosov Moscow State University, 119992, Moscow, Russia.
Protoplasma. 2017 Jan;254(1):557-567. doi: 10.1007/s00709-016-0975-x. Epub 2016 Apr 18.
Cytoplasmic streaming is essential for intracellular communications but its specific functions are not well known. In Chara corallina internodes, long-distance interactions mediated by cyclosis are clearly evident with microscopy-pulse amplitude modulation (PAM) fluorometer under application of localized light (LL) pulses to a remote cell region. Measurements of LL-induced profiles of chlorophyll fluorescence F' at various distances from the LL source suggest that illuminated chloroplasts release into the streaming cytoplasm excess reducing equivalents that are entrained by the fluid flow and transiently reduce the intersystem electron carriers in chloroplasts of downstream shaded areas. The reducing equivalents propagate to distances up to 4.5 mm from the LL source, with the transport rate nearly equal to the velocity of liquid flow. The F' transients disappeared after the arrest of streaming with cytochalasin D and reappeared upon its recovery in washed cells. The F' responses to a distant LL were used as an indicator for the passage of cytosolic reductants across the analyzed cell area during measurements of cell surface pH (pH) in intact and microperforated internodes. In microwounded cell regions, the LL-induced increase in F' occurred synchronously with the increase in pH, by contrast to a slight decrease in pH observed prior to perforation. The results show that reducing agents transported with the cytoplasmic flow are involved in rapid pH changes on the surface of microinjured cells. A possibility is considered that cytoplasmic reductants are processed by stress-activated plasmalemmal NADPH oxidase carrying electrons to oxygen with the eventual H consumption on the outer cell side.
胞质环流对于细胞内通讯至关重要,但其具体功能尚不清楚。在轮藻节间中,在向远端细胞区域施加局部光(LL)脉冲的情况下,通过显微镜脉冲幅度调制(PAM)荧光计可以清楚地观察到由胞质环流介导的长距离相互作用。对距LL光源不同距离处LL诱导的叶绿素荧光F'曲线的测量表明,受光照的叶绿体将过量的还原当量释放到流动的细胞质中,这些还原当量被液流夹带,并短暂还原下游阴影区域叶绿体中的电子传递链中间载体。还原当量传播到距LL光源最远4.5毫米的距离,传输速率几乎等于液流速度。在用细胞松弛素D使环流停止后,F'瞬变消失,在洗涤后的细胞恢复环流时又重新出现。在完整和微穿孔节间测量细胞表面pH(pH)时,对远处LL的F'反应被用作细胞质还原剂穿过分析细胞区域的指标。在微损伤的细胞区域,LL诱导的F'增加与pH增加同步发生,这与穿孔前观察到的pH略有下降形成对比。结果表明,随细胞质流动运输的还原剂参与了微损伤细胞表面的快速pH变化。有一种可能性被认为,细胞质还原剂是由应激激活的质膜NADPH氧化酶处理的,该酶将电子传递给氧气,最终在细胞外侧消耗H。