In this article, the functions of D-myo-inositol-1,4,5-trisphosphate (Ins [1,4,5] P3) in regulating pollen tube polar growth were investigated by application of caged version of the phosphoinositides. To increase the ...In this article, the functions of D-myo-inositol-1,4,5-trisphosphate (Ins [1,4,5] P3) in regulating pollen tube polar growth were investigated by application of caged version of the phosphoinositides. To increase the intracellular Ins（1,4,5）P3 concentration at the apical region of pollen tube, the caged Ins（1,4,5）P3 loaded by osmotic shock was activated by 10 s 360 nm UV flash at this domain （10 μm from the tip）. More than 70% pollen tubes were induced swelling at apex and/or growth axis reorientation, accompanying temporarily growth arrest, by localized increase of Ins（1,4,5）P3 concentration （n=21）. While, pollen tubes without being loaded with caged Ins（1,4,5）P3 had not response to the same dosage UV flash. With FM 1-43 fluorescent staining, it was found that growth perturbation by UV activated caged Ins（1,4,5）P3 had tight link with membrane trafficking at the apical zone of pollen tubes. Upon UV pulse, the apical V-shaped bright area where was full of secretory vesicles spread to a much broader region, which implied that actin filaments at the apical region were remodeled. It was also observed that the FM 1-43 fluorescence intensity at tip remarkably increased than that before UV flash, which demonstrated that more secretory vesicles were accumulated at this region. To estimate the role of Ins（1,4,5）P3 in modulating intracellular calcium concentration and distribution, dextran conjugated fluorescent dye Calcium Green-1 and Rhodamine B were microinjected into pollen tubes together with caged Ins（1,4,5）P3. The results showed that calcium concentration at the subapical region increased upon UV released Ins（1,4,5）P3. Consequently, the tip-focused calcium gradient under the apical dome of pollen tube was disturbed. Simultaneously, the pollen tube bulged at the apical region and its growth rate decreased. As the tip-focused calcium gradient was reestablished, the pollen tube morphology and growth recovered to the normal level. Therefore, Ins（1,4,5）P3 can modulate pollen tube growth rate and direction through mobilizing intracellular calcium and regulating vesicle trafficking during pollen tube finding its way to the ovary.展开更多
本文研究了虾青素对秀丽隐杆线虫衰老和正常生理功能的影响及其相关作用机制。试验设立对照组和虾青素组(0.08 m M、0.16 m M、0.40 m M、0.80 m M),观察N2野生型线虫的寿命及相关生理指标(生殖能力、吞咽频率和移动力),评价不同浓度虾...本文研究了虾青素对秀丽隐杆线虫衰老和正常生理功能的影响及其相关作用机制。试验设立对照组和虾青素组(0.08 m M、0.16 m M、0.40 m M、0.80 m M),观察N2野生型线虫的寿命及相关生理指标(生殖能力、吞咽频率和移动力),评价不同浓度虾青素对线虫衰老过程的影响,明确虾青素作用的最佳剂量;选用最佳剂量的虾青素处理N2野生型、daf-16基因缺陷型和age-1基因缺陷型线虫,分析其寿命长短,探讨虾青素在胰岛素信号通路上延缓线虫衰老的作用机理。结果表明,虾青素可显著延长线虫的寿命(P<0.05),0.16 m M虾青素的作用效果最佳,线虫的平均寿命达到26.03 d,显著延长了31.68%;虾青素对线虫的生殖能力和吞咽频率没有显著影响,能显著增强10 d和15 d线虫的移动力。0.16 m M虾青素可显著提高age-1缺陷型线虫的寿命,对daf-16缺陷型线虫的寿命无显著影响。因此,虾青素可显著延长线虫寿命,其作用机制与胰岛素信号通路上的daf-16基因密切相关。展开更多
文摘In this article, the functions of D-myo-inositol-1,4,5-trisphosphate (Ins [1,4,5] P3) in regulating pollen tube polar growth were investigated by application of caged version of the phosphoinositides. To increase the intracellular Ins（1,4,5）P3 concentration at the apical region of pollen tube, the caged Ins（1,4,5）P3 loaded by osmotic shock was activated by 10 s 360 nm UV flash at this domain （10 μm from the tip）. More than 70% pollen tubes were induced swelling at apex and/or growth axis reorientation, accompanying temporarily growth arrest, by localized increase of Ins（1,4,5）P3 concentration （n=21）. While, pollen tubes without being loaded with caged Ins（1,4,5）P3 had not response to the same dosage UV flash. With FM 1-43 fluorescent staining, it was found that growth perturbation by UV activated caged Ins（1,4,5）P3 had tight link with membrane trafficking at the apical zone of pollen tubes. Upon UV pulse, the apical V-shaped bright area where was full of secretory vesicles spread to a much broader region, which implied that actin filaments at the apical region were remodeled. It was also observed that the FM 1-43 fluorescence intensity at tip remarkably increased than that before UV flash, which demonstrated that more secretory vesicles were accumulated at this region. To estimate the role of Ins（1,4,5）P3 in modulating intracellular calcium concentration and distribution, dextran conjugated fluorescent dye Calcium Green-1 and Rhodamine B were microinjected into pollen tubes together with caged Ins（1,4,5）P3. The results showed that calcium concentration at the subapical region increased upon UV released Ins（1,4,5）P3. Consequently, the tip-focused calcium gradient under the apical dome of pollen tube was disturbed. Simultaneously, the pollen tube bulged at the apical region and its growth rate decreased. As the tip-focused calcium gradient was reestablished, the pollen tube morphology and growth recovered to the normal level. Therefore, Ins（1,4,5）P3 can modulate pollen tube growth rate and direction through mobilizing intracellular calcium and regulating vesicle trafficking during pollen tube finding its way to the ovary.
文摘本文研究了虾青素对秀丽隐杆线虫衰老和正常生理功能的影响及其相关作用机制。试验设立对照组和虾青素组(0.08 m M、0.16 m M、0.40 m M、0.80 m M),观察N2野生型线虫的寿命及相关生理指标(生殖能力、吞咽频率和移动力),评价不同浓度虾青素对线虫衰老过程的影响,明确虾青素作用的最佳剂量;选用最佳剂量的虾青素处理N2野生型、daf-16基因缺陷型和age-1基因缺陷型线虫,分析其寿命长短,探讨虾青素在胰岛素信号通路上延缓线虫衰老的作用机理。结果表明,虾青素可显著延长线虫的寿命(P<0.05),0.16 m M虾青素的作用效果最佳,线虫的平均寿命达到26.03 d,显著延长了31.68%;虾青素对线虫的生殖能力和吞咽频率没有显著影响,能显著增强10 d和15 d线虫的移动力。0.16 m M虾青素可显著提高age-1缺陷型线虫的寿命,对daf-16缺陷型线虫的寿命无显著影响。因此,虾青素可显著延长线虫寿命,其作用机制与胰岛素信号通路上的daf-16基因密切相关。
