Identification and characterization of the bombykal receptor in the hawkmoth Manduca sexta
Identification and characterization of the bombykal receptor in the hawkmoth Manduca sexta
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DOI：
10.1242/jeb.154260
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发表时间：
2017-05
影响因子：
2.8
通讯作者：
D. Wicher;Satoshi Morinaga;Lorena Halty-deLeon;Nico W. Funk;B. Hansson;K. Touhara;M. Stengl
中科院分区：
文献类型：
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作者：
D. Wicher;Satoshi Morinaga;Lorena Halty-deLeon;Nico W. Funk;B. Hansson;K. Touhara;M. Stengl
摘要天蛾雌蛾通过释放一种具有种特异性的混合性信息素来吸引配偶，其中主要成分是（E，Z）-10，12-十六碳二烯醛和（E，E，Z）-10，12，14-十六碳三烯醛。在这里，我们寻找天蛾bombykal受体在异源表达系统。在非洲爪蟾卵母细胞中与MsexOrco共表达的推定信息素受体MsexOr 1在施用家蚕钾（10-300 μmol l−1）后引起剂量依赖性内向电流，并且在HEK 293和CHO细胞中共表达引起细胞内游离Ca 2+浓度的家蚕钾依赖性增加。此外，与MsexOrco共表达的家蚕BmOr 3受体对家蚕（30-100 μmol l−1）产生内向电流。相比之下，与MsexOrco共表达的MsexOr 4对bombykal（30-100 μmol l−1）和（E，E，Z）-10，12，14-十六碳三烯醛模拟物均无反应。因此，MsexOr 1是天蛾中与蚕素结合的信息素受体，而不是MsexOrco，也可能不是MsexOr 4。最后，我们获得的证据表明，磷脂酶C和蛋白激酶C的活性参与天蛾的家蚕受体介导的Ca 2+信号在HEK 293和CHO细胞。总结：受体复合物MsexOr 1 +MsexOrco在异源系统（非洲爪蟾卵母细胞、HEK 293细胞、CHO细胞）中表达时响应性信息素bombykal。
ABSTRACT Manduca sexta females attract their mates with the release of a species-specific sex-pheromone blend, with bombykal (E,Z)-10,12-hexadecadienal and (E,E,Z)-10,12,14-hexadecatrienal being the two major components. Here, we searched for the hawkmoth bombykal receptor in heterologous expression systems. The putative pheromone receptor MsexOr1 coexpressed with MsexOrco in Xenopus oocytes elicited dose-dependent inward currents upon bombykal application (10–300 μmol l−1), and coexpressed in HEK293 and CHO cells caused bombykal-dependent increases in the intracellular free Ca2+ concentration. In addition, the bombykal receptor of Bombyx mori BmOr3 coexpressed with MsexOrco responded to bombykal (30–100 μmol l−1) with inward currents. In contrast, MsexOr4 coexpressed with MsexOrco responded neither to bombykal (30–100 μmol l−1) nor to the (E,E,Z)-10,12,14-hexadecatrienal mimic. Thus, MsexOr1, but not MsexOrco and probably not MsexOr4, is the bombykal-binding pheromone receptor in the hawkmoth. Finally, we obtained evidence that phospholipase C and protein kinase C activity are involved in the hawkmoth's bombykal-receptor-mediated Ca2+ signals in HEK293 and CHO cells. Summary: The receptor complex MsexOr1+MsexOrco responds to the sex pheromone bombykal when expressed in heterologous systems (Xenopus oocytes, HEK293 cells, CHO cells).