Knockin' on pollen's door: live cell imaging of early polarization events in germinating Arabidopsis pollen.
Knockin' on pollen's door: live cell imaging of early polarization events in germinating Arabidopsis pollen.
复制标题
敲开花粉之门：发芽拟南芥花粉早期极化事件的活细胞成像。
DOI：
10.3389/fpls.2015.00246
复制
发表时间：
2015
影响因子：
5.6
通讯作者：
Sprunck S
中科院分区：
文献类型：
--
作者：
Vogler F;Konrad SS;Sprunck S
花粉管是研究细胞动力学和协调极化尖端生长的复杂信号通路的优秀系统。尽管已经描述了作用于尖端生长的花粉管的几种信号机制，但我们对花粉萌发过程中的亚细胞和分子事件以及花粉质膜生长位点选择的了解相当缺乏。为了同时跟踪来自多达 12 种基因不同的植物的发芽花粉，我们开发了一种廉价且易于安装的技术，适用于每种标准显微镜设置。我们通过使用标记花粉细胞质、囊泡、肌动蛋白细胞骨架或精子细胞核和细胞膜的荧光标记线，在拟南芥花粉激活、发芽和花粉管尖端生长建立过程中进行高放大倍数活细胞成像。我们的研究揭示了花粉激活过程中独特的囊泡和 F-肌动蛋白极化以及花粉萌发和花粉管形成过程中的特征生长动力学。最初，发芽的拟南芥花粉管生长缓慢并形成均匀的圆形凸起，随后是一个过渡阶段，囊泡在生长部位大量积累，然后转为快速尖端生长。此外，我们发现在尖端快速生长阶段开始后，两个精子细胞被输送到花粉管中。这里提出的方法适用于定量研究拟南芥花粉萌发和生长过程中的亚细胞事件，以及花粉突变体在花粉质膜上的花粉极化、膨胀或生长位点选择方面的详细分析。
Pollen tubes are an excellent system for studying the cellular dynamics and complex signaling pathways that coordinate polarized tip growth. Although several signaling mechanisms acting in the tip-growing pollen tube have been described, our knowledge on the subcellular and molecular events during pollen germination and growth site selection at the pollen plasma membrane is rather scarce. To simultaneously track germinating pollen from up to 12 genetically different plants we developed an inexpensive and easy mounting technique, suitable for every standard microscope setup. We performed high magnification live-cell imaging during Arabidopsis pollen activation, germination, and the establishment of pollen tube tip growth by using fluorescent marker lines labeling either the pollen cytoplasm, vesicles, the actin cytoskeleton or the sperm cell nuclei and membranes. Our studies revealed distinctive vesicle and F-actin polarization during pollen activation and characteristic growth kinetics during pollen germination and pollen tube formation. Initially, the germinating Arabidopsis pollen tube grows slowly and forms a uniform roundish bulge, followed by a transition phase with vesicles heavily accumulating at the growth site before switching to rapid tip growth. Furthermore, we found the two sperm cells to be transported into the pollen tube after the phase of rapid tip growth has been initiated. The method presented here is suitable to quantitatively study subcellular events during Arabidopsis pollen germination and growth, and for the detailed analysis of pollen mutants with respect to pollen polarization, bulging, or growth site selection at the pollen plasma membrane.
登录
查看更多内容
影响因子：
2.9
作者：
GERVAIS, C;SIMMONDS, DH;NEWCOMB, W
通讯作者：
NEWCOMB, W
影响因子：
6.2
作者：
Daher, Firas Bou;Chebli, Youssef;Geitmann, Anja
通讯作者：
Geitmann, Anja
影响因子：
7.2
作者：
Clough, SJ;Bent, AF
通讯作者：
Bent, AF
影响因子：
7.4
作者：
Cai, Giampiero;Faleri, Claudia;Cresti, Mauro
通讯作者：
Cresti, Mauro
影响因子：
3
作者：
CRESTI, M;CIAMPOLINI, F;MULCAHY, G
通讯作者：
MULCAHY, G