4.6 Article

Molecular characterization of neuropeptide elevenin and two elevenin receptors, IsElevRl and IsElevR2, from the blacklegged tick, Ixodes scapularis

期刊

INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY
卷 101, 期 -, 页码 66-75

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PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.ibmb.2018.07.005

关键词

Salivary gland; GPCR; Deorphanization; Neuropeptide

资金

  1. National Institutes of Health [R01AI090062]

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Understanding salivation in hematophagous arthropod vectors is crucial to developing novel methods to prevent vector-borne disease transmission. The interactions between the tick, host, and pathogens during salivation are highly complex, and are dynamically regulated by the tick central nervous system (synganglion). Recently, tick salivary modulation via neuropeptides was highlighted by mapping neuropeptidergic cells in the synganglion and salivary glands in hard ticks. In this study, we characterized the role of a novel neuropeptide, elevenin (IsElev), and its receptors (IsElevRl and IsElevR2) in the innervation of the salivary glands from Ixodes scapularis female ticks. Homology-based BLAST searches of the I. scapularis genome and Sequence Read Archive (SRA), followed by gene cloning, identified candidate genes: IsElev, IsElevRl, and IsElevR2. The IsElev candidate contained common elevenin features: a signal peptide immediately before an elevenin precursor and two cysteines. During functional assays, synthetic IsElev efficiently activated both IsElevRl and IsElevR2, as indicated by elevated calcium mobilization. IsElevRl (EC50: 0.01 nM) was about 560 times more sensitive to synthetic IsElev than IsElevR2 (EC50: 5.59 nM). Immunoreactivity (IR) for IsElev and IsElevRl was detected as a complex neuronal projection and several neurons in the synganglion. In salivary glands, IsElev-IR was detected in an axonal projection on the surface of the main salivary duct and in axon terminals within type II/Ill salivary gland acini, which are colocalized with SlFamide-IR. IsElevRl-IR was detected on the lumina] surface of both type II/III acini. IsElev transcript levels were high in the synganglion and reached a peak at day 5 post-blood feeding. Salivary glands expressed IsElevRl, which gradually increased over the course of blood feeding until repletion. Here, we propose that IsElev and IsElevRl, localized in salivary gland acini types II/III, are likely involved in tick salivary secretion in the rapid engorgement phase of tick feeding. In addition, we also provide the evidences for IsElev action on the ovary by showing IsElevRl-IR and IsElevR2-IR on the surface of ovary.

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