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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >SUBCELLULAR COMPARTMENTATION OF DIFFERENT LIPOPHILIC FLUORESCEIN DERIVATIVES IN MAIZE ROOT EPIDERMAL CELLS
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SUBCELLULAR COMPARTMENTATION OF DIFFERENT LIPOPHILIC FLUORESCEIN DERIVATIVES IN MAIZE ROOT EPIDERMAL CELLS

机译：玉米根表皮细胞中不同脂多糖荧光素的亚细胞分隔

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Fluorescence microscopy offers some distinct advantages over other techniques for studying ion transport processes in situ with plant cells. However, the use of this technology in plant cells has been limited by our lack of understanding the mechanisms that influence the subcellular distribution of dyes after loading with the lipophilic precursors. In this study, the subcellular distribution of 5-(and 6-)carboxydichlorofluorescein (CDCF), carboxy-SNAFL-1, and carboxy-SNARF-1 was compared to that of 2',7'-bis-(2-carboxyethyl)-5-(and 6-)carboxyfluorescein (BCECF) after incubation of maize roots with their respective lipophilic precursors. Previously, we reported that incubation of roots with BCECF-acetomethyl ester (BCECF-AM) led to vacuolar accumulation of this dye. Similar results were found when roots were incubated with CDCF-diacetate. In contrast, carboxy-SNAFL-1 appeared to be confined to the cytoplasm based on the distribution of fluorescence and the excitation spectra of the dye in situ. On the other hand, incubation of roots with carboxy-SNARF-1-acetoxymethyl acetate yielded fluorescence throughout the cell. When the cytoplasm of epidermal cells was loaded with the BCECF acid by incubation at pH 4 in the absence of external Ca, the dye was retained in the cytoplasm at least 3 h after the loading period. This result indicated that vacuolar accumulation of BCECF during loading of BCECF-AM was not due to transport of BCECF from cytoplasm to vacuole. The esterase activities responsible for the production of either carboxy-SNAFL-1 or BCECF from their respective lipophilic precursor by extracts of roots were compared. The characterization of esterase activities was consistent with the subcellular distribution of these dyes in root cells. The results of these experiments suggest that in maize root epidermal cells the subcellular distribution of these fluorescein dyes may be determined by the characteristics of the esterase activities responsible for hydrolysis of the lipophilic precursor. [References: 25]
机译：与其他技术相比，荧光显微镜在研究植物细胞原位离子迁移过程方面具有一些明显的优势。但是，由于缺乏对影响亲脂性前体负载后染料亚细胞分布的机制的了解，因此限制了该技术在植物细胞中的使用。在这项研究中，比较了5（和6-）羧基二氯荧光素（CDCF），羧基SNAFL-1和羧基SNARF-1与2'，7'-双-（2-羧基乙基）的亚细胞分布玉米根与它们各自的亲脂性前体孵育后的-5-（和6-）羧基荧光素（BCECF）。以前，我们报道了将根与BCECF-乙酰甲酯（BCECF-AM）一起孵育会导致这种染料的液泡积累。当根与CDCF-二乙酸盐一起孵育时，发现了相似的结果。相反，基于荧光的分布和染料的原位激发光谱，羧基-SNAFL-1似乎局限于细胞质。另一方面，将根与羧基-SNARF-1-乙酰氧基乙酸甲酯一起温育会在整个细胞内产生荧光。当通过在不存在外部Ca的情况下在pH 4下孵育将BCECF酸装载到表皮细胞的细胞质中时，染料在装载期后至少3 h保留在细胞质中。该结果表明，在装载BCECF-AM期间，BCECF的液泡积累不是由于BCECF从细胞质向液泡的转运。比较了通过根提取物从它们各自的亲脂性前体产生羧基SNAFL-1或BCECF的酯酶活性。酯酶活性的表征与这些染料在根细胞中的亚细胞分布一致。这些实验的结果表明，在玉米根表皮细胞中，这些荧光素染料的亚细胞分布可通过负责亲脂性前体水解的酯酶活性的特征来确定。 [参考：25]

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