In moths octopamine improved pheromone-dependent mate search time dependently. In the nocturnal hawkmoth Manduca sexta long-term tip recordings of trichoid sensilla were performed to investigate whether biogenic amines modulate pheromone transduction time dependently. At three Zeitgebertimes octopamine, tyramine and the octopamine antagonist epinastine were applied during non-adapting pheromone-stimulation. At ZT 8-11, during the photophase, when sensilla were adapted, octopamine and to a lesser extent tyramine increased the bombykal-dependent sensillar potential amplitude and initial action potential (AP) frequency. In addition, during the photophase, when sensilla are less able to resolve pheromone pulses, octopamine rendered pheromone responses more phasic and sensitive, and raised the spontaneous AP frequency. During the late scotophase, at ZT 22-1, when the antenna appeared maximally sensitized for pheromone pulse detection and endogenous octopamine levels are high, exogenously applied octopamine was ineffective. Epinastine blocked the pheromone-dependent AP response at ZT 8-11 and slightly affected it at ZT 22-1, while it had no effect on the sensillar potential amplitude. Epinastine decreased the spontaneous AP activity during photophase and scotophase and rendered pheromone responses more tonic in the scotophase. We hypothesize that the presence of octopamine in the antenna is obligatory for the detection of intermittent pheromone pulses at all Zeitgebertimes.

SUMMARY Pheromone-dependent mate search is under strict circadian control in different moth species. But it remains unknown whether daytime-dependent changes in pheromone sensitivity already occur at the periphery in male moths. Because adapting pheromone stimuli cause rises of cyclic guanosine monophosphate (cGMP) in pheromone-sensitive trichoid sensilla of the night-active hawkmoth Manduca sexta, we wanted to determine whether cGMP decreases pheromone-sensitivity of olfactory receptor neurons in a daytime-dependent manner. Long-term tip recordings from trichoid sensilla were performed at the early day (ZT 1-4), when many moths are still active, and at the middle of the day (ZT 8-11), when moths are resting. A non-adapting pheromone-stimulation protocol combined with perfusion of the sensillum lymph with the membrane-permeable cGMP analogue 8bcGMP adapted the action potential response but not the sensillar potential. Perfusion with 8bcGMP decreased the initial action potential frequency, decreased the numbers of action potentials elicited in the first 100 ms of the pheromone response and attenuated the reduction of action potential amplitude. Furthermore, the decrease in 8bcGMP-dependent action potential frequency was stronger in recordings made at ZT 8-11 than at ZT 1-4. In the control recordings during the course of the day the pheromone responses became increasingly tonic and less phasic. At ZT 8-11 only, this daytime-dependent effect was further enhanced by 8bcGMP application. Thus we hypothesize that during the moths' resting phase,elevated cGMP levels underlie a daytime-dependent decrease in pheromone sensitivity and a decline in the temporal resolution of pheromone pulses.

SUMMARYOctopamine causes time-dependent disadaptation of pheromone-sensitive olfactory receptor neurons (ORNs) of Manduca sexta. Because the majority of insect octopamine receptors are positively coupled to adenylyl cyclases we examined whether cyclic adenosine monophosphate (cAMP) mimics octopamine-dependent modulation of pheromone transduction in a time-dependent manner. Long-term tip recordings of single trichoid sensilla of Manduca sexta were performed during three zeitgeber times (ZTs, ZT 0=lights on), while stimulating the sensilla with two doses of the main pheromone component bombykal in a non-adapting protocol. The membrane-permeable cAMP analogue 8bcAMP increased the normalized sensillar potential amplitude in a time-and bombykal dose-dependent way. At the higher bombykal dose only, the applied 8bcAMP antagonized an endogenous decrease in the mean sensillar potential amplitude at ZT 1-4 and ZT 8-11 when ORNs were adapted but not at ZT 22-1, when ORNs were sensitized. In contrast to octopamine, 8bcAMP did not consistently affect the initial pheromone-dependent action potential frequency, the phasic/tonic response pattern, or the time-dependent shift to lower mean action potential frequencies at ZT 8-11. Furthermore, 8bcAMP increased the spontaneous action potential frequency time dependently, but differently from octopamine. In conclusion, our results show that cAMP only partly mimics the octopamine-dependent disadaptation of olfactory receptor neurons during photophase, apparently due to another missing octopamine-dependent synergistic factor such as defined intracellular calcium levels. Supplementary material available online at