School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland, Muttenz, Switzerland.
Department of Environmental Systems Science, Institute of Biogeochemistry and Pollution Dynamics, Swiss Federal Institute of Technology Zürich (ETH Zürich), Zürich, Switzerland.
Environ Toxicol Chem. 2019 May;38(5):1074-1083. doi: 10.1002/etc.4383. Epub 2019 Apr 1.
The insect yolk precursor vitellogenin is a lipoglycoprotein synthesized and stored in the fat body and secreted into the hemolymph. In honey bees, vitellogenin displays crucial functions in hormone signaling, behavioral transition of nurse bees to foragers, stress resistance, and longevity in workers. Plant protection products such as neonicotinoids, pyrethroids, and organophosphates alter the transcriptional expression of vitellogenin. To assess plant protection product-induced alterations on the protein level, we developed a rabbit polyclonal vitellogenin antibody. After characterization, we assessed its specificity and vitellogenin levels in different tissues of worker bees. The vitellogenin antibody recognized full-length 180-kDa vitellogenin and the lighter fragment of 150 kDa in fat body, hemolymph, and brain. In hemolymph, a band of approximately 75 kDa was detected. Subsequent mass spectrometric analysis (liquid chromatography-mass spectrometry) confirmed the 180- and 150-kDa bands as vitellogenin. Subsequently, we evaluated vitellogenin expression in brain, fat body, and hemolymph on 24-h exposure of bees to 3 ng/bee to the neonicotinoid clothianidin. Full-length vitellogenin was upregulated 3-fold in the fat body, and the 150-kDa fragment was upregulated in the brain of exposed honey bees, whereas no alteration occurred in the hemolymph. Upregulation of the vitellogenin protein by the neonicotinoid clothianidin is in line with the previously shown induction of its transcript. We conclude that vitellogenin might serve as a potential biomarker for neonicotinoid and other pesticide exposure in bees. Environ Toxicol Chem 2019;00:1-10. © 2019 SETAC.
昆虫卵黄前体卵黄蛋白原是一种糖脂蛋白，在脂肪体中合成和储存，并分泌到血淋巴中。在蜜蜂中，卵黄蛋白原在激素信号转导、工蜂从保姆蜂向觅食蜂的行为转变、抗应激和寿命方面发挥着至关重要的作用。农药如新烟碱类、拟除虫菊酯类和有机磷类会改变卵黄蛋白原的转录表达。为了评估农药对蛋白质水平的诱导变化，我们开发了一种兔多克隆卵黄蛋白原抗体。经过表征，我们评估了它在工蜂不同组织中的特异性和卵黄蛋白原水平。卵黄蛋白原抗体识别全长 180kDa 的卵黄蛋白原和 150kDa 的较轻片段在脂肪体、血淋巴和脑中。在血淋巴中，检测到大约 75kDa 的条带。随后的质谱分析（液相色谱-质谱）证实 180kDa 和 150kDa 带为卵黄蛋白原。随后，我们评估了蜜蜂暴露于 3ng/bee 新烟碱类农药噻虫嗪 24 小时后大脑、脂肪体和血淋巴中的卵黄蛋白原表达。全长卵黄蛋白原在脂肪体中上调 3 倍，150kDa 片段在暴露于蜜蜂的脑中上调，而血淋巴中没有变化。新烟碱类农药噻虫嗪对卵黄蛋白原蛋白的上调与之前报道的其转录物的诱导一致。我们得出结论，卵黄蛋白原可能成为蜜蜂暴露于新烟碱类和其他农药的潜在生物标志物。环境毒理化学 2019;00:1-10。©2019 SETAC。
