NMT历史上的今天丨J Proteome Res：胞外Ca2+内流对松果花粉管发育影响的蛋白质组学和细胞学综合分析

2008年04月01日，中科院植物研究所林金星、吴小琴用NMT在Journal of Proteome Research上发表了标题为Integrative proteomic and cytological analysis of the effects of extracellular Ca2+ influx on Pinus bungeana pollen tube development的研究成果。

• 期刊：J Proteome Res
• 主题：胞外Ca²⁺内流对松果花粉管发育影响的蛋白质组学和细胞学综合分析
• 标题：Integrative proteomic and cytological analysis of the effects of extracellular Ca²⁺ influx on Pinus bungeana pollen tube development
• 影响因子：5.684
• 检测指标：Ca²⁺流速
• 通讯作者：中科院植物研究所林金星、吴小琴

英文摘要

Ca²⁺ is an essential ion in the control of pollen germination and tube growth. However, the control of pollen tube development by Ca²⁺ signaling and its interactions with cytoskeletal components, energy-providing pathways, and cell-expansion machinery remain elusive. Here, we used nifedipine (Nif) to study Ca²⁺ functions in differential protein expression and other cellular processes in Pinus bungeana pollen tube growth.

Proteomics analysis indicated that 50 proteins showed differential expression with varying doses of Nif. Thirty-four of these were homologous to previously reported proteins and were classified into different functional categories closely related to tip-growth machinery.

Blocking the L-type Ca²⁺ channel with Nif in the pollen tube membrane induced several early alterations within a short time, including a reduction of extracellular Ca²⁺ influx and a subsequently dramatic decrease in cytosolic free Ca²⁺ concentration ([Ca²⁺]c), concomitant with ultrastructural abnormalities and changes in the abundance of proteins involved in energy production and signaling. Secondary alterations included actin filament depolymerization, disrupted patterns of endocytosis/exocytosis, and cell wall remodeling, along with changes in the proteins involved in these processes.

These results suggested that extracellular Ca²⁺ influx was necessary for the maintenance of the typical tip-focused [Ca²⁺]c gradient in the P. bungeana pollen tube, and that reduced adenosine triphosphate production (ATP), depolymerization of the cytoskeleton, and abnormal endocytosis/exocytosis, together with enhanced rigidity of cell walls, were responsible for the growth arrest observed in pollen tubes treated with Nif.

 

中文摘要（谷歌机翻）

Ca²⁺是控制花粉萌发和管生长的必需离子。但是，通过Ca²⁺信号传导控制花粉管发育及其与细胞骨架成分，能量提供途径和细胞扩增机制的相互作用仍然难以捉摸。在这里，我们使用硝苯地平（Nifdipine）研究松果花粉花粉管生长中Ca²⁺在差异蛋白表达和其他细胞过程中的功能。

蛋白质组学分析表明，随着Nif剂量的变化，50种蛋白质表现出差异表达。其中的34个与先前报道的蛋白质同源，并被归类为与尖端生长机制密切相关的不同功能类别。

在花粉管膜中用Nif阻断L型Ca²⁺通道可在短时间内引起数个早期变化，包括细胞外Ca²⁺内流减少和随后胞浆游离Ca²⁺浓度（[Ca²⁺] c）急剧下降，并伴有超微结构能量产生和信号传导中涉及的蛋白质异常和丰度变化。次要的变化包括肌动蛋白丝解聚，内吞/胞吐作用的破坏模式，细胞壁重塑以及这些过程中涉及的蛋白质变化。

这些结果表明，胞外Ca²⁺内流对于维持P. bungeana花粉管中典型的尖端集中[Ca²⁺] c梯度是必要的，并且减少了三磷酸腺苷的产生（ATP），细胞骨架的解聚以及异常的内吞/胞吐作用以及细胞壁的刚性增强是造成在用Nif处理的花粉管中观察到的生长停滞的原因。

Figure 1. Dose-dependent inhibitory effects of exogenous Nif on extracellular Ca²⁺ influx at the pollen tube tips. A typical extracellular Ca²⁺ influx can be detected in the very tip of control pollen tube, and 100 µM Nif treatment decreased the Ca²⁺ influx, while 250 and 500 µM Nif treatment greatly reduced this Ca²⁺ influx.

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