Molecular characterization of the vitellogenin receptor from the tick, Amblyomma hebraeum (Acari: Ixodidae).

We have identified the full-length cDNA encoding a vitellogenin receptor (VgR) from the African bont tick Amblyomma hebraeum Koch (1844). VgRs are members of the low-density lipoprotein receptor superfamily that promote the uptake of the yolk protein vitellogenin (Vg), from the haemolymph. The AhVgR (GenBank accession No. JX846592) is 5703 bp, and encodes an 1801 aa protein with a 196.5 kDa molecular mass following cleavage of a 22 aa signal peptide. Phylogenetic analysis indicates that AhVgR is highly similar to other tick VgRs. AhVgR is expressed in only the ovary of mated, engorged females, and is absent in all other female tissues and in both fed and unfed males. Unfed, adult females injected with a VgR-dsRNA probe to knock-down VgR expression experienced a significant delay in ovary development and started oviposition significantly later than controls. These results indicate that the expression of AhVgR is important for the uptake of Vg and subsequent maturation of the oocytes.

Ticks: physiological aspects with implications for pathogen transmission.

Ticks have attracted a great deal of scientific attention primarily because of their role as vectors of numerous pathogens. The majority of tick researchers worldwide focus primarily on microbiological and clinical issues relating to these pathogens, and on methods (pesticidal and biological) for controlling tick populations. Unfortunately, it is often forgotten that ticks are also interesting in their own right to the general biologist because of their unusual physiological (and other) adaptations. Here I review some of these adaptations relating primarily to osmoregulation. (i) I outline their ability to take up water vapour directly from the atmosphere, an adaptation that enables them to withstand desiccation for extended periods while unfed and, in the case of larvae and nymphs, following engorgement. (ii) I present the remarkable filtration-resorption mechanism of the argasid tick coxal organ, analogous to that of the vertebrate glomerular kidney, that enables them to regulate haemolymph fluid volume and composition following the blood meal. (iii) I then turn attention to the salivary glands of female ixodid ticks, which serve the on-host osmoregulatory function in this family of ticks, (iv) and I discuss the pharmacological control of salivary fluid secretion. (v) Finally, I link the latter to the mechanism of pathogen transmission by the salivary glands, using the tick-borne Thogoto virus as a specific example.

Identification of two genes essential for sperm development in the male tick Amblyomma hebraeum Koch (Acari: Ixodidae).

In most ticks of the family Ixodidae, gonad maturation and spermatogenesis are stimulated by the taking of a blood meal. Previous work from this laboratory identified 35 genes that are up-regulated by feeding [Weiss, B.L., Stepczynski, J.M., Wong, P., Kaufman, W.R., 2002. Identification and characterization of genes differentially expressed in the testis/vas deferens of the fed male tick, Amblyomma hebraeum. Insect Biochemistry and Molecular Biology 32, 785-793]. The functions of most of these genes remain unknown. We used RNA interference technology to investigate the consequences of blocking the function of 13 of these genes. Attenuation of the expression of two of these in particular, AhT/VD 8 and AhT/VD 10, correlated with deformities in the testis and abnormalities in spermiogenesis. Furthermore, most females fed in the company of these males did not engorge properly and laid many fewer eggs, most of which were infertile.

Gluttony and sex in female ixodid ticks: how do they compare to other blood-sucking arthropods?

The central issue dealt with here is the role of copulation in the control of feeding behaviour in ticks and some haematophagous insects. Female ticks of the family Ixodidae normally engorge to approximately 100 x their unfed body weight, and then drop from the host, produce and lay eggs, and die. Virgins, on the other hand, normally do not exceed 5-40% (depending on species) of the normal engorged body weight. But instead of detaching voluntarily at that point most virgins remain fixed to the host for extended periods, waiting for males to find them so they can complete engorgement. Virgin haematophagous insects, and virgin ticks of the family Argasidae display little, if any, reduction in blood meal size compared to mated females, at least not during the first ovarian cycle. During subsequent ovarian cycles, meal size in some virgin insects may be somewhat reduced depending on how many eggs are retained in the reproductive tract, but the reduction is not nearly to the same extent as that observed for virgin ixodid females. The stimulatory effect of copulation on engorgement in the latter is caused by a pair of proteins (voraxin alpha and beta) produced in the testis and transferred to the female with the spermatophore. Here, I propose why it might be adaptive for an ixodid female to remain small until mated. The hypothesis is suggested from the facts that ixodid ticks remain attached to the host for days (rather than minutes), and that virgin ticks, above a certain critical weight, lose all opportunity for producing viable offspring should they be groomed off the host prematurely, or should the host die while ticks are still attached.

Quantification of vitellogenesis and its control by 20-hydroxyecdysone in the ixodid tick, Amblyomma hebraeum.

Ovaries of the ixodid tick, Amblyomma hebraeum Koch, grew rapidly after engorgment as a result of yolk uptake. At 26 degrees C, oviposition began by day 10 post-engorgement, plateaued on days 16-18, and ended by day 38. Vitellin (Vt) was partially purified from ovaries of day 10 engorged ticks by gel filtration and ion exchange chromatography. This Vt comprises seven major and several minor polypeptides. Two polypeptides (211 and 148 kD) from haemolymph of engorged female ticks corresponded to minor polypeptides of similar molecular weight in the ovary. The haemolymph titre of the 211 and 148 kD polypeptides increased up to the onset of oviposition. These polypeptides were absent in males and non-vitellogenic females (day 0 engorged or day 10 partially-fed females), and were thus designated as vitellogenin (Vg). Antibodies raised against haemolymph Vg211 and 148 recognized these polypeptides in partially purified Vt, as well as six of the seven major polypeptides. Using these antibodies we developed an indirect, competitive ELISA to quantify Vg. Rise in haemolymph Vg-concentration lagged slightly behind the rise in haemolymph ecdysteroid (ES)-concentration, and Vg-synthesis was stimulated by injections of 20E into non-vitellogenic females. These observations indicate that an ES is the vitellogenic hormone in A. hebraeum.

The relationship between 'critical weight' and 20-hydroxyecdysone in the female ixodid tick, Amblyomma hebraeum.

THE FEEDING CYCLE OF ADULT FEMALE TICKS (ACARI: Ixodidae) is divided into preparatory, slow and rapid feeding phases. At the transition from slow to rapid feeding, Amblyomma hebraeum (Koch) females reach a 'critical weight' (CW; approx. 10x the unfed weight) that is characterized by several behavioural and physiological changes. Five of these changes were used as criteria to establish a more precise estimate of CW than we have to date. The CW as defined by re-attachment to the host was 9x the unfed weight, while for haemolymph ecdysteroid titre, salivary gland degeneration, ovary weight, oocyte length and oocyte vitellin content the CW was 10x, 10x, 12x, 12x and 13x, respectively. CW thus varies depending on the parameter measured. Although previous studies have established the influence of ecdysteroids on salivary gland degeneration and vitellogenesis, here we demonstrate a further effect of ecdysteroids: inhibiting re-attachment to the host.