Serotonin modulates insect hemocyte phagocytosis via two different serotonin receptors.

Serotonin (5-HT) modulates both neural and immune responses in vertebrates, but its role in insect immunity remains uncertain. We report that hemocytes in the caterpillar, Pieris rapae are able to synthesize 5-HT following activation by lipopolysaccharide. The inhibition of a serotonin-generating enzyme with either pharmacological blockade or RNAi knock-down impaired hemocyte phagocytosis. Biochemical and functional experiments showed that naive hemocytes primarily express 5-HT1B and 5-HT2B receptors. The blockade of 5-HT1B significantly reduced phagocytic ability; however, the blockade of 5-HT2B increased hemocyte phagocytosis. The 5-HT1B-null Drosophila melanogaster mutants showed higher mortality than controls when infected with bacteria, due to their decreased phagocytotic ability. Flies expressing 5-HT1B or 5-HT2B RNAi in hemocytes also showed similar sensitivity to infection. Combined, these data demonstrate that 5-HT mediates hemocyte phagocytosis through 5-HT1B and 5-HT2B receptors and serotonergic signaling performs critical modulatory functions in immune systems of animals separated by 500 million years of evolution.

De novo assembly and characterization of central nervous system transcriptome reveals neurotransmitter signaling systems in the rice striped stem borer, Chilo suppressalis.

BACKGROUND: Neurotransmitter signaling systems play crucial roles in multiple physiological and behavioral processes in insects. Genome wide analyses of de novo transcriptome sequencing and gene specific expression profiling provide rich resources for studying neurotransmitter signaling pathways. The rice striped stem borer, Chilo suppressalis is a destructive rice pest in China and other Asian countries. The characterization of genes involved in neurotransmitter biosynthesis and transport could identify potential targets for disruption of the neurochemical communication and for crop protection. RESULTS: Here we report de novo sequencing of the C. suppressalis central nervous system transcriptome, identification and expression profiles of genes putatively involved in neurotransmitter biosynthesis, packaging, and recycling/degradation. A total of 54,411 unigenes were obtained from the transcriptome analysis. Among these unigenes, we have identified 32 unigenes (31 are full length genes), which encode 21 enzymes and 11 transporters putatively associated with biogenic aminergic signaling, acetylcholinergic signaling, glutamatergic signaling and GABAergic signaling. RT-PCR and qRT-PCR results indicated that 12 enzymes were highly expressed in the central nervous system and all the transporters were expressed at significantly high levels in the central nervous system. In addition, the transcript abundances of enzymes and transporters in the central nervous system were validated by qRT-PCR. The high expression levels of these genes suggest their important roles in the central nervous system. CONCLUSIONS: Our study identified genes potentially involved in neurotransmitter biosynthesis and transport in C. suppressalis and these genes could serve as targets to interfere with neurotransmitter production. This study presents an opportunity for the development of specific and environmentally safe insecticides for pest control.

Larvae of the small white butterfly, Pieris rapae, express a novel serotonin receptor.

The biogenic amine serotonin (5-hydroxytryptamine, 5-HT) is a neurotransmitter in vertebrates and invertebrates. It acts in regulation and modulation of many physiological and behavioral processes through G-protein-coupled receptors. Five 5-HT receptor subtypes have been reported in Drosophila that share high similarity with mammalian 5-HT1A, 5-HT1B, 5-HT2A, 5-HT2B, and 5-HT7 receptors. We isolated a cDNA (Pr5-HT8 ) from larval Pieris rapae, which shares relatively low similarity to the known 5-HT receptor classes. After heterologous expression in HEK293 cells, Pr5-HT8 mediated increased [Ca(2+)]i in response to low concentrations (< 10 nM) of 5-HT. The receptor did not affect [cAMP]i even at high concentrations (> 10 µM) of 5-HT. Dopamine, octopamine, and tyramine did not influence receptor signaling. Pr5-HT8 was also activated by various 5-HT receptor agonists including 5-methoxytryptamine, (±)-8-Hydroxy-2-(dipropylamino) tetralin, and 5-carboxamidotryptamine. Methiothepin, a non-selective 5-HT receptor antagonist, activated Pr5-HT8 . WAY 10635, a 5-HT1A antagonist, but not SB-269970, SB-216641, or RS-127445, inhibited 5-HT-induced [Ca(2+)]i increases. We infer that Pr5-HT8 represents the first recognized member of a novel 5-HT receptor class with a unique pharmacological profile. We found orthologs of Pr5-HT8 in some insect pests and vectors such as beetles and mosquitoes, but not in the genomes of honeybee or parasitoid wasps. This is likely to be an invertebrate-specific receptor because there were no similar receptors in mammals. We isolated a cDNA (Pr5-HT8) from larval Pieris rapae, which shares relatively low similarity to the known GPCRs. After heterologous expression in HEK293 cells, Pr5-HT8 mediated increased [Ca(2+)]i in response to low concentrations (< 10 nM) of 5-HT and various 5-HT receptor agonists. We found orthologs of Pr5-HT8 in some insect pests and vectors such as beetles and mosquitoes, but not in the genomes of honeybee, parasitoid wasps, or mammals.