Synganglion transcriptome and developmental global gene expression in adult females of the American dog tick, Dermacentor variabilis (Acari: Ixodidae).

454 Pyrosequencing was used to characterize the expressed genes from the synganglion and associated neurosecretory organs of unfed and partially fed virgin and mated replete females of the American dog tick, Dermacentor variabilis. A total of 14,881 contiguous sequences (contigs) was assembled, with an average size of 229 bp. Gene ontology terms for Level 2 biological processes were assigned to 4366 contigs. Seven acetylcholinesterases, a muscarinic acetylcholine (ACh) receptor, two nicotinic ACh receptor ß-subunits, two ACh unc-18 regulators, two dopamine receptors, two gamma aminobutyric acid (GABA) receptors, two GABA transporters, two norepinephrine transporters and an octopamine receptor are described. Microarrays were conducted to examine global gene expression and quantitative real-time polymerase chain reaction was used to verify expression of selected neuropeptides. Hierarchical clustering of all differentially expressed transcripts grouped part-fed and replete ticks as being more similar in terms of differentially expressed genes with unfed ticks as the outgroup. Nine putative neuropeptides (allatostatin, bursicon-ß, preprocorazonin, glycoprotein hormone α, insulin-like peptide, three orcokinins, preprosulphakinin) and a gonadotropin releasing hormone receptor were differentially expressed, and their developmental expression and role in reproduction was investigated. The presence of eclosion hormone, corazonin and bursicon in the synganglion, which in insects regulate behaviour and cuticle development associated with moulting, suggest that this system may be used in ticks to regulate blood feeding, cuticle expansion and development related to female reproduction; adult ticks do not moult.

Molecular characterization of the major hemelipoglycoprotein in ixodid ticks.

The major hemelipoglyco-carrier protein (CP) found throughout the development of male and female adult American dog ticks, Dermacentor variabilis (Say) was sequenced. DvCP is a single transcript coding for two protein subunits that together contain three motifs: (1) a lipoprotein n-terminal domain that is a common attribute of proteins that bind lipids, carbohydrates and metals; (2) a domain of unknown function characteristic of proteins with several large open beta sheets; and (3) a von Willebrand factor type D domain near the carboxy-terminus apparently important for multimerization. These motifs, which are also found in tick vitellogenin, are not shared by heme-binding proteins studied thus far in other hematophagous insects. DvCP message was highest in fat body and salivary gland but was also found in midgut and ovary tissue. Expression was initiated by blood feeding in virgin females and not by mating, as is typical of tick vitellogenin; and the message was found in fed males at levels similar to part fed, virgin females. CP appears to be highly conserved among the Ixodida. The closest match by BlastP to DvCP is vitellogenin from Caenorhabditis elegans (AAC04423), suggesting that CP is a novel protein. The role of CP in heme sequestration, the evolution of hematophagy and host complementation are discussed.