Effects of sex and time of day on metabolism and excretion of corticosterone in urine and feces of mice
Effects of sex and time of day on metabolism and excretion of corticosterone in urine and feces of mice
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DOI：
10.1016/s0016-6480(02)00620-2
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发表时间：
2003-02-15
影响因子：
2.7
通讯作者：
Palme, R
中科院分区：
文献类型：
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作者：
Touma, C;Sachser, N;Palme, R
用于监测小鼠等小动物应激激素的非侵入性技术具有多种优势，并且在实验室和现场研究中都有很高的需求。由于有关糖皮质激素的物种特异性代谢和排泄的知识对于开发这种技术至关重要，因此我们在小鼠（Mus musculus f. Domesticus，菌株 C57BL/6J）中进行了放射代谢研究。每只小鼠腹腔注射 740 kBq H-3 标记的皮质酮，并收集五天内所有排泄的尿液和粪便样本。在第一个实验中，16 只动物（每种性别 8 只）在上午 9 点接受注射，而 8 只小鼠（每种性别 4 只）在晚上 9 点接受注射。在第二个实验中。在这两项实验中，放射性代谢物主要在粪便中回收，尽管男性通过粪便排出的比例（约 73%）明显高于女性（约 53%）。注射后约2小时内检测到尿液中的放射性峰值，而粪便中的放射性峰值浓度随后观察到（取决于注射时间：实验1中注射后约10小时，实验2中注射后约4小时，从而证明了一天中时间的影响）。通过高效液相色谱 (HPLC) 测定粪便 [H-3] 皮质酮代谢物的数量和相对丰度。 HPLC 分离显示皮质酮主要被广泛代谢为极性更强的物质。关于形成的代谢物的类型，男性和女性之间存在显着差异，但实验之间没有显着差异。此外，通过使用四种酶免疫测定 (EIA) 筛选 HPLC 级分来评估这些代谢物的免疫反应性。然而，只有新建立的 5alpha-pregnane-3beta, 11beta,21-triol-20-one（测量具有 5alpha-3beta, 11beta-diol 结构的皮质酮代谢物）的 EIA 在两性中检测到几个高强度的放射性代谢物峰，而其他 EIA 仅显示出轻微的免疫反应性。因此，我们的研究首次提供了有关小鼠尿液和粪便中皮质酮代谢和排泄的大量信息，并且首次证明了动物性别和一天中时间的显着影响。基于这些数据，应该可以通过使用新开发的 EIA 测量粪便皮质酮代谢物来非侵入性地监测该物种的肾上腺皮质活性。由于小鼠在世界范围内的研究中被广泛使用，这可能会为从生态学到行为内分泌学的各个领域开辟新的视角。 (C) 2003 年爱思唯尔科学（美国）。版权所有。
Non-invasive techniques to monitor stress hormones in small animals like mice offer several advantages and are highly demanded in laboratory as well as in field research. Since knowledge about the species-specific metabolism and excretion of glucocorticoids is essential to develop such a technique, we conducted radiometabolism studies in mice (Mus musculus f. domesticus, strain C57BL/6J). Each mouse was injected intraperitoneally with 740 kBq of H-3-labelled corticosterone and all voided urine and fecal samples were collected for five days. In a first experiment 16 animals (eight of each sex) received the injection at 9 a.m., while eight mice (four of each sex) were injected at 9 p.m. in a second experiment. In both experiments radioactive metabolites were recovered predominantly in the feces, although males excreted significantly higher proportions via the feces (about 73%) than females (about 53%). Peak radioactivity in the urine was detected within about 2 h after injection, while in the feces peak concentrations were observed later (depending on the time of injection: about 10 h postinjection in experiment 1 and about 4 h postinjection in experiment 2, thus proving an effect of the time of day). The number and relative abundance of fecal [H-3]corticosterone metabolites was determined by high performance liquid chromatography (HPLC). The HPLC separations revealed that corticosterone was extensively metabolized mainly to more polar substances. Regarding the types of metabolites formed, significant differences were found between males and females, but not between the experiments. Additionally, the immunoreactivity of these metabolites was assessed by screening the HPLC fractions with four enzyme immunoassays (EIA). However, only a newly established EIA for 5alpha-pregnane-3beta, 11beta,21-triol-20-one (measuring corticosterone metabolites with a 5alpha-3beta, 11beta-diol structure) detected several peaks of radioactive metabolites with high intensity in both sexes, while the other EIAs showed only minor immunoreactivity. Thus, our study for the first time provides substantial information about metabolism and excretion of corticosterone in urine and feces of mice and is the first demonstrating a significant impact of the animals' sex and the time of day. Based on these data it should be possible to monitor adrenocortical activity non-invasively in this species by measuring fecal corticosterone metabolites with the newly developed EIA. Since mice are extensively used in research world-wide, this could open new perspectives in various fields from ecology to behavioral endocrinology. (C) 2003 Elsevier Science (USA). All rights reserved.