Oviposition is induced upon mating in most insects. Ovulation is a primary step in oviposition, representing an important target to control insect pests and vectors, but limited information is available on the underlying mechanism. Here we report that the beta adrenergic-like octopamine receptor Octβ2R serves as a key signaling molecule for ovulation and recruits protein kinase A and Ca2+/calmodulin-sensitive kinase II as downstream effectors for this activity. We found that the octβ2r homozygous mutant females are sterile. They displayed normal courtship, copulation, sperm storage and post-mating rejection behavior but were unable to lay eggs. We have previously shown that octopamine neurons in the abdominal ganglion innervate the oviduct epithelium. Consistently, restored expression of Octβ2R in oviduct epithelial cells was sufficient to reinstate ovulation and full fecundity in the octβ2r mutant females, demonstrating that the oviduct epithelium is a major site of Octβ2R’s function in oviposition. We also found that overexpression of the protein kinase A catalytic subunit or Ca2+/calmodulin-sensitive protein kinase II led to partial rescue of octβ2r’s sterility. This suggests that Octβ2R activates cAMP as well as additional effectors including Ca2+/calmodulin-sensitive protein kinase II for oviposition. All three known beta adrenergic-like octopamine receptors stimulate cAMP production in vitro. Octβ1R, when ectopically expressed in the octβ2r’s oviduct epithelium, fully reinstated ovulation and fecundity. Ectopically expressed Octβ3R, on the other hand, partly restored ovulation and fecundity while OAMB-K3 and OAMB-AS that increase Ca2+ levels yielded partial rescue of ovulation but not fecundity deficit. These observations suggest that Octβ2R have distinct signaling capacities in vivo and activate multiple signaling pathways to induce egg laying. The findings reported here narrow the knowledge gap and offer insight into novel strategies for insect control.

Aminergic signaling modulates associative learning and memory. Substantial advance has been made in Drosophila on the dopamine receptors and circuits mediating olfactory learning; however, our knowledge of other aminergic modulation lags behind. To address this knowledge gap, we investigated the role of octopamine in olfactory conditioning. Here, we report that octopamine activity through the b-adrenergic-like receptor Octb1R drives aversive and appetitive learning: Octb1R in the mushroom body ab neurons processes aversive learning, whereas Octb1R in the projection neurons mediates appetitive learning. Our genetic interaction and imaging studies pinpoint cAMP signaling as a key downstream effector for Octb1R function. The rutabaga-adenylyl cyclase synthesizes cAMP in a Ca 21 /calmodulin-dependent manner, serving as a coincidence detector for associative learning and likely representing a downstream target for Octb1R. Supporting this notion, the double heterozygous rutabaga/1;octb1r/1 flies perform poorly in both aversive and appetitive conditioning, while individual heterozygous rutabaga/1 and octb1r/1 flies behave like the wild-type control. Consistently, the mushroom body and projection neurons in the octb1r brain exhibit blunted responses to octopamine when cAMP levels are monitored through the cAMP sensor. We previously demonstrated the pivotal functions of the D 1 receptor dDA1 in aversive and appetitive learning, and the a1 adrenergic-like receptor OAMB in appetitive learning. As expected, octb1r genetically interacts with dumb (dDA1 mutant) in aversive and appetitive learning, but it interacts with oamb only in appetitive learning. This study uncovers the indispensable contributions of dopamine and octopamine signaling to aversive and appetitive learning. All experiments were performed on mixed sex unless otherwise noted.

Active forgetting is an essential component of the brain’s memory management system 1 . Forgetting can be permanent, in which prior memory is lost completely; or transient, in which memory exists in a temporary state of impaired retrieval. Such temporary blocks on memory seem universal, and can disrupt an individual’s plans, social interactions, and ability to make rapid, flexible and appropriate choices. However, the neurobiological mechanisms that cause transient forgetting are unknown. Here we identify a single dopamine neuron in Drosophila that mediates memory suppression resulting in transient forgetting. Artificially activating this neuron failed to abolish the expression of long-term memory. Rather, it briefly suppressed memory retrieval, with memory becoming accessible with time. The dopamine neuron modulates memory retrieval by stimulating a unique dopamine receptor expressed in a restricted physical compartment of the axons of mushroom body neurons. This mechanism for transient forgetting is triggered by interfering stimuli presented just prior to retrieval.

Forgetting is an essential component of the brain’s memory management system, providing a balance to memory formation processes by removing unused or unwanted memories, or by suppressing their expression. However, the molecular, cellular, and circuit mechanisms underlying forgetting are poorly understood. Here we show that the memory suppressor gene, sickie , functions in a single dopamine neuron (DAn) by supporting the process of active forgetting in Drosophila . RNAi knockdown (KD) of sickie impairs forgetting by reducing the Ca 2+ influx and DA release from the DAn that promotes forgetting. Coimmunoprecipitation/mass spectrometry analyses identified cytoskeletal and presynaptic active zone (AZ) proteins as candidates that physically interact with Sickie, and a focused RNAi screen of the candidates showed that Bruchpilot (Brp)—a presynaptic AZ protein that regulates calcium channel clustering and neurotransmitter release—impairs active forgetting like sickie KD. In addition, overexpression of brp rescued the impaired forgetting of sickie KD, providing evidence that they function in the same process. Moreover, we show that sickie KD in the DAn reduces the abundance and size of AZ markers but increases their number, suggesting that Sickie controls DAn activity for forgetting by modulating the presynaptic AZ structure. Our results identify a molecular and circuit mechanism for normal levels of active forgetting and reveal a surprising role of Sickie in maintaining presynaptic AZ structure for neurotransmitter release.