Cholecystokinin-like peptide mediates satiety by inhibiting sugar attraction.

Feeding is essential for animal survival and reproduction and is regulated by both internal states and external stimuli. However, little is known about how internal states influence the perception of external sensory cues that regulate feeding behavior. Here, we investigated the neuronal and molecular mechanisms behind nutritional state-mediated regulation of gustatory perception in control of feeding behavior in the brown planthopper and Drosophila. We found that feeding increases the expression of the cholecystokinin-like peptide, sulfakinin (SK), and the activity of a set of SK-expressing neurons. Starvation elevates the transcription of the sugar receptor Gr64f and SK negatively regulates the expression of Gr64f in both insects. Interestingly, we found that one of the two known SK receptors, CCKLR-17D3, is expressed by some of Gr64f-expressing neurons in the proboscis and proleg tarsi. Thus, we have identified SK as a neuropeptide signal in a neuronal circuitry that responds to food intake, and regulates feeding behavior by diminishing gustatory receptor gene expression and activity of sweet sensing GRNs. Our findings demonstrate one nutritional state-dependent pathway that modulates sweet perception and thereby feeding behavior, but our experiments cannot exclude further parallel pathways. Importantly, we show that the underlying mechanisms are conserved in the two distantly related insect species.

Pymetrozine inhibits reproductive behavior of brown planthopper Nilaparvata lugens and fruit fly Drosophila melanogaster.

Pymetrozine is a promising chemical used to control brown planthopper, which developed resistance to imidacloprid and buprofezin in the past decade. Field efficacy indicates that pymetrozine can reduce the number of offsprings of brown planthopper, but the specific physiological mechanism is unknown. In this study, we systematically described the mating process of brown planthopper including 8 steps (abdominal vibration, following, positioning, wing extension, attempted copulation, copulation, terminated copulation and leaving) and explored the optimal mating time after adult eclosion (3-5 days) and observation time (30 mins). Also, behavioral data showed that pymetrozine can affect the mating behavior and female fecundity of brown planthopper and fruit fly. As one of the target genes for pymetrozine, Nanchung (Nan), the nan36a mutant male courtship index, female receptivity and the number of offsprings were significantly decreased. Behavioral defects in nan36a mutant flies can be rescued by expressed NlNan. Our results indicated that Nan plays essential roles in the mating behavior and female fecundity. These findings provide useful information for demonstrating that pymetrozine effectively reduce the reproduction of brown planthopper and contribute to our understanding of reproductive strategies controlled by pymetrozine in insects.

Pymetrozine activates TRPV channels of brown planthopper Nilaparvata lugens.

The commercial insecticide pymetrozine has been extensively used for brown planthopper control in East Asia. The transient receptor potential vanilloid (TRPV) channel, which consists of two proteins, Nanchung (Nan) and Inactive (Iav), has recently been shown to be the molecular target of pymetrozine in the fruit fly (Drosophila melanogaster) and pea aphid (Acyrthosiphon pisum). In this study, we characterized the Nan and Iav TRPV channel subunits of N. lugens and measured the action of pymetrozine on them. NlNan and NlIav are structurally similar to homologs from other insects. The expression pattern analysis of various body parts showed that NlNan and NlIav were both more abundantly expressed in antennae. When NlNan and NlIav were co-expressed in Xenopus laevis oocytes, they formed channels with high sensitivity to pymetrozine (EC50 = 5.5 × 10-8 M). Behavioral observation revealed that the gravitaxis defect in the fruit fly nan36a mutant was rescued by ectopically expressed NlNan and the rescued behavior could be abolished by pymetrozine. Our results confirm that NlNan and NlIav co-expressed complexes can be activated by pymetrozine both in vitro and in vivo and provide useful information for future resistance mechanism studies.

Mechanism of metabolic resistance to pymetrozine in Nilaparvata lugens: over-expression of cytochrome P450 CYP6CS1 confers pymetrozine resistance.

BACKGROUND: Pymetrozine is commonly used for the control of Nilaparvata lugens, and resistance to pymetrozine has been frequently reported in the field populations in recent years. However, the mechanism of brown planthopper resistance to pymetrozine is still unknown. RESULTS: In this study, a pymetrozine-resistant strain (PMR) was established, and the potential biochemical resistance mechanism of N. lugens to pymetrozine was investigated. Pymetrozine was synergized by the inhibitor piperonyl butoxide (PBO) in the PMR with 2.83-fold relative synergistic ratios compared with the susceptible strain (Sus). Compared with the Sus, the cytochrome P450 monooxygenase activity of PMR was increased by 1.7 times, and two P450 genes (NlCYP6CS1 and NlCYP301B1) were found to be significantly overexpressed more than 6.0-fold in the PMR. Pymetrozine exposure induced upregulation of NlCYP6CS1 expression in the Sus, but the expression of NlCYP301B1 did not change significantly. In addition, RNA interference (RNAi)-mediated suppression of NlCYP6CS1 gene expression dramatically increased the toxicity of pymetrozine against N. lugens. Moreover, transgenic lines of Drosophila melanogaster expressing NlCYP6CS1 were less susceptible to pymetrozine, and had a stronger ability to metabolize pymetrozine. CONCLUSIONS: Taken together, our findings indicate that the overexpression of NlCYP6CS1 is one of the key factors contributing to pymetrozine resistance in N. lugens. And this result is helpful in proposing a management strategy for pymetrozine resistance.

A Mechanosensory Receptor TMC Regulates Ovary Development in the Brown Planthopper Nilaparvata lugens.

Transmembrane channel-like (TMC) genes encode a family of evolutionarily conserved membrane proteins. Mutations in the TMC1 and TMC2 cause deafness in humans and mice. However, their functions in insects are is still not well known. Here we cloned three tmc genes, Nltmc3, Nltmc5, and Nltmc7 from brown planthoppers. The predicted amino acid sequences showed high identity with other species homologs and have the characteristic eight or nine transmembrane domains and TMC domain architecture. We detected these three genes in all developmental stages and examined tissues. Interestingly, we found Nltmc3 was highly expressed in the female reproductive organ especially in the oviduct. RNAi-mediated silencing of Nltmc3 substantially decreased the egg-laying number and impaired ovary development. Our results indicate that Nltmc3 has an essential role in the ovary development of brown planthoppers.

The NompC channel regulates Nilaparvata lugens proprioception and gentle-touch response.

NompC channel is a member of the transient receptor potential (TRP) ion channel superfamily. It can regulate gentle-touch, locomotion, hearing and food texture detection in Drosophila. We cloned the NompC gene of Nilaparvata lugens (NlNompC). The full length NlNompC possessed similar structure as DmNompC, which belongs to TRPN subfamily. The expression pattern analysis of different developmental stages and body parts showed that the transcription of NlNompC was more abundant in adult stage and in the abdomen. Injection of double-stranded RNA (dsRNA) of NlNompC in the third-instar nymphs successfully knocked down the target gene with 75% suppression. At nine days after injection, the survival rate of dsRNA injected nymphs was as low as 9.84%. Behavioral observation revealed that the locomotion of the dsRNA injected nymphs was defective with much less movement compared to the negative control. Feeding and honeydew excretion of the dsRNA injected insects also decreased significantly. These results suggested that NlNompC is a classical mechanotransduction channel that plays important roles in proprioception and locomotion, and is essential for the survival of N. lugens. The results also contribute to the understanding of how TRP channels regulate proprioception.