Differential ammonia metabolism in Aedes aegypti fat body and midgut tissues.
Differential ammonia metabolism in Aedes aegypti fat body and midgut tissues.
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DOI：
10.1016/j.jinsphys.2010.02.016
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发表时间：
2010-09
影响因子：
2.2
通讯作者：
Miesfeld, Roger L.
中科院分区：
文献类型：
--
作者：
Scaraffia, Patricia Y.;Zhang, Qingfen;Thorson, Kelsey;Wysocki, Vicki H.;Miesfeld, Roger L.
为了了解在组织水平上埃及伊蚊如何应对有毒的氨浓度，从血粉蛋白质的快速代谢的结果，我们调查了15 N从15 NH 4Cl到氨基酸的掺入使用在体外组织培养系统。将来自雌性蚊子的脂肪体或中肠组织在补充有葡萄糖和15 NH 4Cl的伊蚊盐水溶液中孵育10-40分钟。然后将培养基与氘标记的氨基酸混合，干燥并衍生化。通过质谱技术定量每个样品中的15 N-标记和未标记的氨基酸。结果表明，这两种组织有效地将氨纳入氨基酸，然而，具体的代谢途径是不同的。在脂肪体中，来自15 NH 4Cl的15 N首先结合到Gln的酰胺侧链中，然后结合到Gln、Glu、Ala和Pro的氨基中。该过程主要通过谷氨酰胺合成酶（GS）和谷氨酸合成酶（GltS）途径发生。相比之下，中肠中的15 N首先掺入Glu和Ala的氨基，然后掺入Gln的酰胺侧链。有趣的是，我们的数据显示GS/GltS途径在中肠中不起作用。相反，中肠细胞通过谷氨酸脱氢酶、丙氨酸氨基转移酶和GS来解毒氨。这些数据提供了新的见解氨代谢在A。埃及蚊子。
In order to understand at the tissue level how Aedes aegypti copes with toxic ammonia concentrations that result from the rapid metabolism of blood meal proteins, we investigated the incorporation of 15N from 15NH4Cl into amino acids using an in vitro tissue culture system. Fat body or midgut tissues from female mosquitoes were incubated in an Aedes saline solution supplemented with glucose and 15NH4Cl for 10–40 minutes. The media was then mixed with deuterium-labeled amino acids, dried and derivatized. The 15N-labeled and unlabeled amino acids in each sample were quantified by mass spectrometry techniques. The results demonstrate that both tissues efficiently incorporate ammonia into amino acids, however, the specific metabolic pathways are distinct. In the fat body, the 15N from 15NH4Cl is first incorporated into the amide side chain of Gln and then into the amino group of Gln, Glu, Ala and Pro. This process mainly occurs via the glutamine synthetase (GS) and glutamate synthase (GltS) pathway. In contrast, 15N in midgut is first incorporated into the amino group of Glu and Ala, and then into the amide side chain of Gln. Interestingly, our data show that the GS/GltS pathway is not functional in the midgut. Instead, midgut cells detoxify ammonia by glutamate dehydrogenase, alanine aminotransferase and GS. These data provide new insights into ammonia metabolism in A. aegypti mosquitoes.
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影响因子：
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