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首页> 外文期刊>The Journal of Experimental Biology >Ion-selective microelectrode analysis of salicylate transport by the Malpighian tubules and gut of Drosophila melanogaster
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Ion-selective microelectrode analysis of salicylate transport by the Malpighian tubules and gut of Drosophila melanogaster

机译：果蝇小肠和马尔皮基小管和肠道对水杨酸转运的离子选择性微电极分析

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摘要

Transport of the organic anion salicylate by the Malpighian tubules and gut of larval and adult fruit flies was studied using two salicylate-selective microelectrode methods. The first method combined the high selectivity of tridodecylmethylammonium-based electrodes for salicylate with the self-referencing ion-selective microelectrode technique for non-invasive spatial and temporal analysis of salicylate flux. Measurements with this technique revealed secretion of salicylate across the main and distal segments of the Malpighian tubule as well as the midgut, ileum and rectum. The second method used a salicylate-selective microelectrode to measure the concentration of salicylate in fluid droplets secreted by isolated Drosophila Malpighian tubules set up in a Ramsay secretion assay. Transepithelial salicylate flux was calculated as the product of fluid secretion rate and secreted fluid salicylate concentration. Measurements with this method revealed that salicylate transport was active and saturable; the kinetic parameters J(max) and K(t) were 2.72 pmol min(-1) tubule(-1) and 0.046 mmol l(-1), respectively. Measurements of transepithelial salicylate flux determined by both microelectrode methods were in good agreement. Transepithelial flux measurements measured by microelectrodes were also validated by comparing them with measurements of radiolabelled salicylate levels in secreted droplets. Salicylate concentrations in haemolymph samples were measured with salicylate-selective microelectrodes after injection of salicylate into the haemocoel or after insects were fed salicylate-rich diets. The rate of salicylate secretion by Malpighian tubules in vitro was sufficient to account for the measured rate of decline of salicylate concentration in the haemolymph in vivo.
机译：使用两种水杨酸盐选择性微电极方法研究了Malpighian小管和幼虫及成年果蝇肠道对有机水杨酸阴离子的转运。第一种方法将三十二烷基甲基铵基电极对水杨酸盐的高选择性与自参考离子选择性微电极技术相结合，用于水杨酸通量的非侵入性时空分析。用该技术进行的测量表明，水杨酸盐在马氏小管的主要和远端以及中肠，回肠和直肠中均分泌。第二种方法使用水杨酸盐选择性微电极来测量在Ramsay分泌分析中建立的分离的果蝇Malpighian小管分泌的液滴中水杨酸盐的浓度。经上皮水杨酸盐通量计算为液体分泌速率与分泌的水杨酸盐浓度的乘积。用这种方法进行的测量表明，水杨酸盐的运输是活跃且可饱和的。动力学参数J（max）和K（t）分别为2.72 pmol min（-1）小管（-1）和0.046 mmol l（-1）。通过两种微电极方法测定的跨上皮水杨酸盐通量的测量结果吻合良好。通过将微电极测量的上皮通量测量值与分泌液滴中放射性标记的水杨酸盐水平的测量值进行比较，也可以验证其有效性。在将水杨酸盐注射入血细胞中后，或在给昆虫喂食富含水杨酸盐的饮食后，用水杨酸盐选择性微电极测量血淋巴样品中的水杨酸盐浓度。 Malpighian小管在体外分泌水杨酸盐的速率足以说明体内血淋巴中水杨酸盐浓度下降的测量速率。

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