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NMT歷*的今天丨胞外Ca2+內流對松果花粉管發育影響的蛋白質組學
NMT作為生命科學底層核心技術，是建立活體創新科研平臺的*技術。2005年~2020年，NMT已扎根中國15年。2020年，中國NMT銷往瑞士蘇黎世大學，正式打開歐洲市場
NMT歷*的今天
2008年04月01日，中科院植物研究所林金星、吳小琴用NMT在Journal of Proteome Research上發表了標題為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
主題：胞外Ca2+內流對松果花粉管發育影響的蛋白質組學和細胞學綜合分析
標題：Integrative proteomic and cytological analysis of the effects of extracellular Ca2+ influx on Pinus bungeana pollen tube development
影響因子：5.684
檢測指標：Ca2+流速
作者：中科院植物研究所林金星、吳小琴
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（谷歌機翻）
Ca2+是控制花粉萌發和管生長的必需離子。但是，通過Ca2+信號傳導控制花粉管發育及其與細胞骨架成分，能量提供途徑和細胞擴增機制的相互作用仍然難以捉摸。在這里，我們使用硝苯地平（Nifdipine）研究松果花粉花粉管生長中Ca2+在差異蛋白表達和其他細胞過程中的功能。
蛋白質組學分析表明，隨著Nif劑量的變化，50種蛋白質表現出差異表達。其中的34個與先前報道的蛋白質同源，并被歸類為與生長機制密切相關的不同功能類別。
在花粉管膜中用Nif阻斷L型Ca2+通道可在短時間內引起數個早期變化，包括細胞外Ca2+內流減少和隨后胞漿游離Ca2+濃度（[Ca2+] c）急劇下降，并伴有超微結構能量產生和信號傳導中涉及的蛋白質異常和豐度變化。次要的變化包括肌動蛋白絲解聚，內吞/胞吐作用的破壞模式，細胞壁重塑以及這些過程中涉及的蛋白質變化。
這些結果表明，胞外Ca2+內流對于維持P. bungeana花粉管中典型的集中[Ca2+] c梯度是必要的，并且減少了三磷酸腺苷的產生（ATP），細胞骨架的解聚以及異常的內吞/胞吐作用以及細胞壁的剛性增強是造成在用Nif處理的花粉管中觀察到的生長停滯的原因。
Figure 1. Dose-dependent inhibitory effects of exogenous Nif on extracellular Ca2+in?ux at the pollen tube tips. A typical extracellular Ca2+ in?ux can be detected in the very tip of control pollen tube, and 100 µM Nif treatment decreased the Ca2+ in?ux, while 250 and 500 µM Nif treatment greatly reduced this Ca2+ in?ux.