首页> 外文OA文献 >Fluorescent n-3 and n-6 Very Long Chain Polyunsaturated Fatty Acids: THREE-PHOTON IMAGING IN LIVING CELLS EXPRESSING LIVER FATTY ACID-BINDING PROTEIN*
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Fluorescent n-3 and n-6 Very Long Chain Polyunsaturated Fatty Acids: THREE-PHOTON IMAGING IN LIVING CELLS EXPRESSING LIVER FATTY ACID-BINDING PROTEIN*

机译:荧光n-3和n-6超长链多不饱和脂肪酸:表达肝脂肪酸结合蛋白的活细胞中的三光子成像*

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摘要

Despite the considerable beneficial effects of n-3 and n-6 very long chain polyunsaturated fatty acids (VLC-PUFAs), very little is known about the factors that regulate their uptake and intracellular distribution in living cells. This issue was addressed in cells expressing liver-type fatty acid-binding protein (L-FABP) by real time multiphoton laser scanning microscopy of novel fluorescent VLC-PUFAs containing a conjugated tetraene fluorophore near the carboxyl group and natural methylene-interrupted n-3 or n-6 grouping. The fluorescent VLC-PUFAs mimicked many properties of their native nonfluorescent counterparts, including uptake, distribution, and metabolism in living cells. The unesterified fluorescent VLC-PUFAs distributed either equally in nuclei versus cytoplasm (22-carbon n-3 VLC-PUFA) or preferentially to cytoplasm (20-carbon n-3 and n-6 VLC-PUFAs). L-FABP bound fluorescent VLC-PUFA with affinity and specificity similar to their nonfluorescent natural counterparts. Regarding n-3 and n-6 VLC-PUFA, L-FABP expression enhanced uptake into the cell and cytoplasm, selectively altered the pattern of fluorescent n-6 and n-3 VLC-PUFA distribution in cytoplasm versus nuclei, and preferentially distributed fluorescent VLC-PUFA into nucleoplasm versus nuclear envelope, especially for the 22-carbon n-3 VLC-PUFA, correlating with its high binding by L-FABP. Multiphoton laser scanning microscopy data showed for the first time VLC-PUFA in nuclei of living cells and suggested a model, whereby L-FABP facilitated VLC-PUFA targeting to nuclei by enhancing VLC-PUFA uptake and distribution into the cytoplasm and nucleoplasm.
机译:尽管n-3和n-6非常长链的多不饱和脂肪酸(VLC-PUFAs)具有相当大的有益作用,但对于调节它们在活细胞中的摄取和细胞内分布的调节因子知之甚少。通过实时多光子激光扫描显微镜对新型荧光VLC-PUFA进行处理,该细胞表达肝脏型脂肪酸结合蛋白(L-FABP),该新型荧光VLC-PUFA含有靠近羧基的共轭四烯荧光团和天然亚甲基干扰的n-3或n-6个分组。荧光VLC-PUFA模仿了其天然非荧光对应物的许多特性,包括在活细胞中的吸收,分布和代谢。未酯化的荧光VLC-PUFAs在细胞核与细胞质中均等分布(22个碳n-3 VLC-PUFA),或者在细胞质中优先分布(20个碳n-3和n-6个VLC-PUFAs)。 L-FABP结合荧光VLC-PUFA的亲和力和特异性与非荧光天然类似物相似。关于n-3和n-6 VLC-PUFA,L-FABP表达增强了对细胞和细胞质的摄取,选择性改变了荧光质n-6和n-3 VLC-PUFA在细胞质相对于细胞核中的分布模式,并优先分布了荧光VLC-PUFA进入核质与核被膜的关系,特别是对于22碳n-3 VLC-PUFA,与其通过L-FABP的高结合力有关。多光子激光扫描显微镜数据首次显示了活细胞核中的VLC-PUFA并提出了一种模型,其中L-FABP通过增强VLC-PUFA的摄取和分布进入细胞质和核质,促进了VLC-PUFA靶向细胞核。

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