CRISPR/Cas 9-Mediated Mutations as a New Tool for Studying Taste in Honeybees
CRISPR/Cas 9-Mediated Mutations as a New Tool for Studying Taste in Honeybees
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DOI：
10.1093/chemse/bjaa063
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发表时间：
2020-10-01
期刊：
影响因子：
3.5
通讯作者：
Scheiner, Ricarda
中科院分区：
文献类型：
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作者：
Degirmenci, Laura;Geiger, Dietmar;Scheiner, Ricarda
蜜蜂以花蜜为主要的碳水化合物来源。蔗糖、葡萄糖和果糖是植物花蜜的主要成分。有趣的是，蜜蜂只表达3种假定的糖受体（AmGr1， AmGr2和AmGr3），这与许多其他昆虫和脊椎动物形成鲜明对比。糖受体仅被部分表征。AmGrl检测不同的糖，包括蔗糖和葡萄糖。AmGr2被认为仅作为一个共受体，而AmGr3被认为是一个果糖受体。我们表明，当非洲爪蟾卵母细胞表达时，蜜蜂味觉受体AmGr3是高度专门化的果糖感知。当我们使用CRISPR/Cas9在雌性工蜂的卵中引入相应的AmGr3基因无义突变时，产生的突变体几乎完全丧失了对果糖的反应性。相反，对蔗糖的反应是正常的。因此，CRISPR/ Cas9在蜜蜂中引入的无义突变可以诱导可测量的行为变化，并用于表征体内味觉受体的功能。CRISPR/Cas9是表征蜜蜂体内味觉受体的一种极好的新工具。非洲爪蟾卵母细胞的生物物理受体特征和蜜蜂AmGr3的无义突变明确表明，该受体对果糖具有高度特异性。
Honeybees rely on nectar as their main source of carbohydrates. Sucrose, glucose, and fructose are the main components of plant nectars. Intriguingly, honeybees express only 3 putative sugar receptors (AmGr1, AmGr2, and AmGr3), which is in stark contrast to many other insects and vertebrates. The sugar receptors are only partially characterized. AmGrl detects different sugars including sucrose and glucose. AmGr2 is assumed to act as a co-receptor only, while AmGr3 is assumedly a fructose receptor. We show that honeybee gustatory receptor AmGr3 is highly specialized for fructose perception when expressed in Xenopus oocytes. When we introduced nonsense mutations to the respective AmGr3 gene using CRISPR/Cas9 in eggs of female workers, the resulting mutants displayed almost a complete loss of responsiveness to fructose. In contrast, responses to sucrose were normal. Nonsense mutations introduced by CRISPR/ Cas9 in honeybees can thus induce a measurable behavioral change and serve to characterize the function of taste receptors in vivo. CRISPR/Cas9 is an excellent novel tool for characterizing honeybee taste receptors in vivo. Biophysical receptor characterization in Xenopus oocytes and nonsense mutation of AmGr3 in honeybees unequivocally demonstrate that this receptor is highly specific for fructose.