Department of Biology, University of Ottawa, Ottawa, ON, Canada.
Mol Cell Endocrinol. 2012 Nov 25;364(1-2):113-25. doi: 10.1016/j.mce.2012.08.017. Epub 2012 Sep 1.
Unlike other freshwater fish previously examined, zebrafish are capable of increasing their rate of Na(+) uptake during chronic exposure to acidic water (pH 4). In the present study, the potential role of cortisol in the induction of Na(+) uptake during acid-exposure was investigated. When zebrafish larvae (4 days post-fertilization) were treated with waterborne cortisol, the rate of Na(+) uptake was significantly increased; this effect was blocked by co-incubating larvae with RU-486, an antagonist selective for the glucocorticoid receptor (GR). A similar induction in Na(+) uptake, which was also blocked by RU-486, was observed when larvae were treated with dexamethasone, a selective GR agonist. Conversely, treating larvae with aldosterone, a selective agonist for the mineralocorticoid receptor (MR) had no effect on Na(+) uptake. Acid-exposure increased whole body cortisol levels and translational knockdown of GR using antisense morpholinos prevented the full induction of Na(+) uptake during exposure to acidic water, further confirming the role of cortisol and GR in Na(+) uptake stimulation. Using immunohistochemistry, GR was localized to ionocytes known to be responsible for Na(+) uptake (HR-cells). Knockdown of Rhcg1, an apical membrane ammonia channel or Na(+)/H(+) exchanger 3b (NHE3b), proteins known to play an important role in facilitating Na(+) uptake in acidic water, prevented the stimulatory effects of cortisol treatment on Na(+) uptake, suggesting that cortisol regulates Na(+) uptake by stimulating an Rhcg1-NHE3b "functional metabolon".
与之前研究的其他淡水鱼类不同,斑马鱼能够在长期暴露于酸性水(pH4)的情况下增加其钠离子摄取率。在本研究中,研究了皮质醇在酸性暴露期间诱导钠离子摄取中的潜在作用。当斑马鱼幼虫(受精后 4 天)用皮质醇处理时,钠离子摄取率显著增加;这种作用被 RU-486 阻断,RU-486 是一种对糖皮质激素受体(GR)具有选择性的拮抗剂。当幼虫用地塞米松(一种选择性 GR 激动剂)处理时,也观察到类似的钠离子摄取诱导,该诱导也被 RU-486 阻断。相反,用醛固酮(一种选择性盐皮质激素受体(MR)激动剂)处理幼虫对钠离子摄取没有影响。酸性暴露增加了全身皮质醇水平,使用反义形态发生素对 GR 进行翻译敲低可防止在暴露于酸性水期间完全诱导钠离子摄取,进一步证实了皮质醇和 GR 在钠离子摄取刺激中的作用。通过免疫组织化学,GR 被定位到已知负责钠离子摄取的离子细胞(HR 细胞)。敲低 Rhcg1,一种顶端膜氨通道或 Na+/H+交换器 3b(NHE3b),这些蛋白在促进酸性水中钠离子摄取中起着重要作用,可防止皮质醇处理对钠离子摄取的刺激作用,表明皮质醇通过刺激 Rhcg1-NHE3b“功能代谢物”来调节钠离子摄取。