Syed Zainulabeuddin, Ishida Yuko, Taylor Katherine, Kimbrell Deborah A, Leal Walter S
Department of Entomology, University of California, Davis, CA 95616, USA.
Proc Natl Acad Sci U S A. 2006 Oct 31;103(44):16538-43. doi: 10.1073/pnas.0607874103. Epub 2006 Oct 23.
We have expressed a male-specific, pheromone-sensitive odorant receptor (OR), BmorOR1, from the silkworm moth Bombyx mori in an "empty neuron" housed in the ab3 sensilla of a Drosophila Deltahalo mutant. Single-sensillum recordings showed that the BmorOR1-expressing neurons in the transgenic flies responded to the B. mori pheromone bombykol, albeit with low sensitivity. These transgenic flies responded to lower doses of bombykol in an altered stimulation method with direct delivery of pheromone into the sensillum milieu. We also expressed a B. mori pheromone-binding protein, BmorPBP, in the BmorOR1-expressing ab3 sensilla. Despite the low levels of BmorPBP expression, flies carrying both BmorOR1 and BmorPBP showed significantly higher electrophysiological responses than BmorOR1 flies. Both types of BmorOR1-expressing flies responded to bombykol, and to a lesser extent to a second compound, bombykal, even without the addition of organic solvents to the recording electrode buffer. When the semiochemicals were delivered by the conventional puffing of stimulus on the antennae, the receptor responded to bombykol but not to bombykal. The onset of response was remarkably slow, and neural activity extended for an unusually long time (>1 min) after the end of stimulus delivery. We hypothesize that BmorOR1-expressing ab3 sensilla lack a pheromone-degrading enzyme to rapidly inactivate bombykol and terminate the signal. We also found an endogenous receptor in one of the sensillum types on Drosophila antenna that responds to bombykol and bombykal with sensitivity comparable to the pheromone-detecting sensilla on B. mori male antennae.
我们在果蝇Delta halo突变体的ab3感器中的“空神经元”中表达了家蚕雄特异性、对信息素敏感的气味受体(OR)BmorOR1。单感器记录显示,转基因果蝇中表达BmorOR1的神经元对家蚕信息素蚕蛾醇有反应,尽管灵敏度较低。这些转基因果蝇在将信息素直接递送到感器环境的改变刺激方法中,对较低剂量的蚕蛾醇有反应。我们还在表达BmorOR1的ab3感器中表达了家蚕信息素结合蛋白BmorPBP。尽管BmorPBP表达水平较低,但同时携带BmorOR1和BmorPBP的果蝇显示出比仅表达BmorOR1的果蝇显著更高的电生理反应。两种表达BmorOR1的果蝇都对蚕蛾醇有反应,在较小程度上也对第二种化合物蚕蛾醛有反应,即使在记录电极缓冲液中不添加有机溶剂也是如此。当通过传统的向触角吹气施加刺激物来递送这些化学信号时,该受体对蚕蛾醇有反应,但对蚕蛾醛没有反应。反应的起始非常缓慢,并且在刺激物递送结束后神经活动持续异常长的时间(>1分钟)。我们推测,表达BmorOR1的ab3感器缺乏一种信息素降解酶来快速使蚕蛾醇失活并终止信号。我们还在果蝇触角的一种感器类型中发现了一种内源性受体,它对蚕蛾醇和蚕蛾醛的反应灵敏度与家蚕雄虫触角上检测信息素的感器相当。