Department of Biological Sciences, Institut de Recherche en Biologie Végétale, University of Montreal, Montreal, Quebec H1X 2B2, Canada.
Traffic. 2011 Nov;12(11):1537-51. doi: 10.1111/j.1600-0854.2011.01256.x. Epub 2011 Aug 22.
In order to accurately target the embryo sac and deliver the sperm cells, the pollen tube has to find an efficient path through the pistil and respond to precise directional cues produced by the female tissues. Although many chemical and proteic signals have been identified to guide pollen tube growth, the mechanism by which the tube changes direction in response to these signals is poorly understood. We designed an experimental setup using a microscope-mounted galvanotropic chamber that allowed us to induce the redirection of in vitro pollen tube growth through a precisely timed and calibrated external signal. Actin destabilization, reduced calcium concentration in the growth medium and inhibition of calcium channel activity decreased the responsiveness of the pollen tube to a tropic trigger. An increased calcium concentration in the medium enhanced this response and was able to rescue the effect of actin depolymerization. Time-lapse imaging revealed that the motion pattern of vesicles and the dynamics of the subapical actin array undergo spatial reorientation prior to the onset of a tropic response. Together these results suggest that the precise targeting of the delivery of new wall material represents a key component in the growth machinery that determines directional elongation in pollen tubes.
为了准确地靶向胚囊并输送精子细胞,花粉管必须在雌组织产生的精确方向信号的指引下,找到一条有效的途径穿过雌蕊。虽然已经确定了许多化学和蛋白质信号来指导花粉管的生长,但花粉管如何根据这些信号改变方向的机制还知之甚少。我们设计了一个使用显微镜安装的电迁移室的实验装置,该装置允许我们通过精确定时和校准的外部信号诱导体外花粉管生长的重新定向。肌动蛋白的不稳定性、生长介质中钙离子浓度的降低和钙通道活性的抑制降低了花粉管对向性触发的反应性。介质中钙离子浓度的增加增强了这种反应,并且能够挽救肌动蛋白解聚的效果。延时成像显示,在向性反应开始之前,囊泡的运动模式和亚顶 actin 阵列的动力学经历了空间重定向。这些结果表明,新壁材料的精确靶向输送代表了决定花粉管定向伸长的生长机制中的一个关键组成部分。