Methods to Test Endocrine Disruption in Drosophila melanogaster
Methods to Test Endocrine Disruption in Drosophila melanogaster
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DOI：
10.3791/59535
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发表时间：
2019-07-01
影响因子：
1.2
通讯作者：
Digilio, Filomena Anna
中科院分区：
文献类型：
--
作者：
Bovier, Tiziana Francesca;Cavaliere, Daniels;Digilio, Filomena Anna
近年来，越来越多的证据表明，所有有机体和环境都暴露在类似激素的化学物质中，这种化学物质被称为内分泌干扰物化学物质(EDCs)。这些化学物质可能会改变内分泌系统的正常平衡，导致不良影响，以及人类种群中荷尔蒙紊乱的数量增加，或者野生动物物种的生长受扰和繁殖减少。对于一些EDC，有记录的健康影响和对其使用的限制。然而，对于他们中的大多数人来说，仍然没有这种意义上的科学证据。为了验证一种化学物质在整个生物体中的潜在内分泌效应，我们需要在适当的模型系统中测试它，以及在果蝇--黑腹果蝇身上进行测试。在这里，我们报道了研究果蝇内分泌干扰的详细体内方案，解决了EDC对果蝇繁殖力/生育力、发育时间和寿命的影响。在过去的几年里，我们利用这些果蝇的生命特征来研究17-α-乙炔雌二醇(EE2)、双酚A(BPA)和双酚AF(BPA F)对果蝇的影响。总而言之，这些检测涵盖了果蝇的所有生命阶段，并使评估所有激素介导的过程中的内分泌干扰成为可能。繁殖力/生育力和发育时序分析分别可用来衡量EDC对果蝇繁殖性能和发育阶段的影响。最后，寿命测试包括对成年人长期接触EDC，并测量他们的存活率。然而，这些生活特征也可能受到几个必须仔细控制的实验因素的影响。因此，在这项工作中，我们建议了一系列程序，我们已经优化了这些分析的正确结果。这些方法使科学家能够确定果蝇中任何EDC或不同EDC的混合物的内分泌干扰，尽管要确定导致这种影响的内分泌机制，可能需要更多的论文。
In recent years there has been growing evidence that all organisms and the environment are exposed to hormone-like chemicals, known as endocrine disruptor chemicals (EDCs). These chemicals may alter the normal balance of endocrine systems and lead to adverse effects, as well as an increasing number of hormonal disorders in the human population or disturbed growth and reduced reproduction in the wildlife species. For some EDCs, there are documented health effects and restrictions on their use. However, for most of them, there is still no scientific evidence in this sense. In order to verify potential endocrine effects of a chemical in the full organism, we need to test it in appropriate model systems, as well as in the fruit fly, Drosophila melanogaster. Here we report detailed in vivo protocols to study endocrine disruption in Drosophila, addressing EDC effects on the fecundity/fertility, developmental timing, and lifespan of the fly. In the last few years, we used these Drosophila life traits to investigate the effects of exposure to 17-alpha-ethinylestradiol (EE2), bisphenol A (BPA), and bisphenol AF (BPA F). Altogether, these assays covered all Drosophila life stages and made it possible to evaluate endocrine disruption in all hormone-mediated processes. Fecundity/fertility and developmental timing assays were useful to measure the EDC impact on the fly reproductive performance and on developmental stages, respectively. Finally, the lifespan assay involved chronic EDC exposures to adults and measured their survivorship. However, these life traits can also be influenced by several experimental factors that had to be carefully controlled. So, in this work, we suggest a series of procedures we have optimized for the right outcome of these assays. These methods allow scientists to establish endocrine disruption for any EDC or for a mixture of different EDCs in Drosophila, although to identify the endocrine mechanism responsible for the effect, further essays could be needed.