Wildling Stefan, Kerschbaum Hubert H
Division of Animal Physiology, Department of Cellular Biology, University of Salzburg, 5020 Salzburg, Austria.
J Comp Physiol B. 2007 May;177(4):401-11. doi: 10.1007/s00360-006-0139-y. Epub 2007 Jan 9.
In the present study, we quantified the physiological consequences of nitric oxide (NO) on ammonium release in tadpoles of Xenopus laevis. Tadpoles exposed to S-nitro-N-acetylpenicillamine (SNAP), an NO-donor, or L: -arginine, the substrate of NO synthase (NOS), showed a reversible decrease, whereas animals exposed to the NOS inhibitor Nomega-methyl-L: -arginine (L: -NMMA) exhibited an increase in ammonium release. Release of ammonium may be of physiological relevance during stress response of the animal. Handling of tadpoles as well as exposure to hyposmotic environments increased ammonium release. To localize NO synthesizing cells, we used diaminofluorescein-diacetate (DAF-2DA), an NO-sensitive fluorescent dye, and NADPH-diaphorase histochemistry, an indicator for NOS activity. We observed a fluorescence signal as well as NADPH-diaphorase activity in small, solitary cells in the epidermis. Similarly to NADPH-diaphorase histochemistry, silver nitrate staining and rhodamine labelling, markers for mitochondria-rich cells, showed a strong reaction in these cells. These observations indicate that NO (1) inhibits ammonium release, and (2) is endogenously synthesized in mitochondria-rich cells in Xenopus tadpoles. Based on our histochemical results, we speculate that gill epithelium and epidermis work in parallel to release ammonium as epidermal tissue contains mitochondria-rich and NADPH-diaphorase positive cells.
在本研究中,我们量化了一氧化氮(NO)对非洲爪蟾蝌蚪铵释放的生理影响。暴露于NO供体S-亚硝基-N-乙酰青霉胺(SNAP)或NO合酶(NOS)底物L-精氨酸的蝌蚪,铵释放呈可逆性降低,而暴露于NOS抑制剂Nω-甲基-L-精氨酸(L-NMMA)的动物铵释放增加。在动物应激反应过程中,铵的释放可能具有生理相关性。处理蝌蚪以及暴露于低渗环境都会增加铵的释放。为了定位NO合成细胞,我们使用了对NO敏感的荧光染料二氨基荧光素二乙酸酯(DAF-2DA)以及作为NOS活性指标的NADPH-黄递酶组织化学方法。我们在表皮中的小的单个细胞中观察到了荧光信号以及NADPH-黄递酶活性。与NADPH-黄递酶组织化学方法类似,作为富含线粒体细胞标记物的硝酸银染色和罗丹明标记在这些细胞中显示出强烈反应。这些观察结果表明,NO(1)抑制铵释放,并且(2)在非洲爪蟾蝌蚪富含线粒体的细胞中内源性合成。基于我们的组织化学结果,我们推测鳃上皮和表皮协同作用释放铵,因为表皮组织含有富含线粒体且NADPH-黄递酶阳性的细胞。