Female Tick Amblyomma Research Articles

Microarray analysis of gene expression changes in feeding female and male lone star ticks, Amblyomma americanum (L)

A collection of EST clones from female tick Amblyomma americanum salivary glands was hybridized to RNA from different feeding stages of female tick salivary glands and from unfed or feeding adult male ticks. In the female ticks, the expression patterns changed dramatically upon starting feeding, then changed again towards the end of feeding. On beginning feeding, genes possibly involved in survival on the host increased in expression as did many housekeeping genes. As feeding progressed, some of the survival genes were downregulated, while others were upregulated. When the tick went into the rapid feeding phase, many of the survival genes were downregulated, while a number of transport-associated genes and genes possibly involved in organ degeneration increased. In the males, the presence of females during feeding made a small difference, but feeding made a larger difference. Males showed clear differences from females in expression, as well. Protein synthesis genes were expressed more in all male groups than in the partially fed females, while the putative secreted genes involved in avoiding host defenses were expressed less.

Ecdysteroid Regulation of Salivary Gland Degeneration in the Ixodid Tick,Amblyomma hebraeum:A Reconciliation ofin Vivoandin VitroObservations

Salivary gland degeneration in the female tickAmblyomma hebraeumKoch is triggered by an ecdysteroid (ES) hormone. Under bothin vivoandin vitroconditions, degeneration requires 4 days for completion. In partially fed females that have fed beyond a “critical weight,” the commitment period for salivary gland degeneration occurs between 24 and 48 h after removal from the host. Although tissue degeneration begins within 24 h postengorgement, ES titer as measured by radioimmunoassay (RIA) does not rise to threshold levels until 48 h postengorgement. To explain this anomaly we examined two hypotheses: (1) there is an early hormonal signal (e.g., 3-dehydroecdysone; 3DE) that is an ES not detectable by the antibody used in our RIA; and (2) the low hemolymph titer during the first 2 days postengorgement is not an accurate reflection of the ES concentration within the tissue itself. 3-Oxoecdysteroid 3β-reductase (ketoreductase) was present in salivary glands, but neither ketoreductase nor 3DE was detected in hemolymph. The ES concentration of salivary gland homogenates was similar to that of hemolymph, while that of saliva was undetectable. Together, these results support our second hypothesis that the metabolically active tissue of the salivary gland experiences a suprathreshold concentration of hormone even though the concentration in hemolymph is below threshold levels.

Further investigations on the action of ecdysteroids on the salivary glands of the female tickAmblyomma americanum

Two phytecdysteroids (abutasterone, makisterone A) and five synthetic ecdysteroid analogues, all at 1 microgram/ml, were tested on salivary glands from the female tick, Amblyomma americanum L. (Acari:Ixodidae), held in organ culture for four days. All of these substances caused a significant reduction in fluid secretory competence of salivary glands in vitro. This constitutes further evidence that the structural requirements for causing salivary-gland degeneration in ticks are similar to those generally required for ecdysteroid activity in other arthropods. Although vertebrate steroids are known to augment fluid secretory competence by salivary glands in organ culture, beta-estradiol was not able to attenuate the degenerating effect of 20-hydroxyecdysone, supporting the suggestion that ecdysteroids and vertebrate steroids have distinct sites/mechanisms of action on this tissue.

Fine structure of the foveae dorsales and foveal glands in the female tick Amblyomma americanum (acari: Ixodoidea: Ixodidae).

The fine structure of the paired foveae dorsales and foveal glands in the unfed female Amblyomma americanum is described and compared with that in other tick species. Each fovea opens to the exterior via pores which lead internally into a single cuticular tube, the pore-tube. This is surrounded by 2-3 epithelial cells. The pore-tube enlarges basally forming a large cavity possessing a cup-shaped cytoplasmic protrusion. The pore-tube cuticular lining extends downward to form an electron-dense, flaplike protrusion bracketing narrow cytoplasmic extensions filled with microtubules. These extensions form a previously undescribed valvelike structure that seems to control the flow of pheromone secretion from the foveal gland to the pore-tube. The single foveal gland lying beneath each pore-tube is composed of 2-3 inner, large, storage cells surrounded by outer, spindle-shaped cells; both types of cells have a characteristic feature of epithelia involved in secretory activity and ion transport. The outer cells extend upward to join the base of the poretube cells by septate desmosomes. A nerve, the foveal nerve, containing axons with neurosecretory vesicles occurs in the vicinity of the foveal gland. The secretory activity of the pheromone glands seems to be partially, if not entirely, under a neural regulation.

Metabolism of ecdysteroids in the female tick Amblyomma hebraeum (Ixodoidea, Ixodidae): accumulation of free ecdysone and 20-hydroxyecdysone in the eggs.

[3H]-20-hydroxyecdysone ([3H]-20E) injected into Amblyomma hebraeum females 7 days before the beginning of oviposition, viz. at the beginning of vitellogenesis, was converted to 3 polar peaks of unknown nature called 1, 2 and 3, and to apolar conjugates AP1, AP2 and AP3. AP2 have the same retention times as the esters of 20E with long chain fatty acids described in Ornithodoros moubata (Diehl et al. 1985). However, principally unmetabolized 20E was incorporated into the ovaries, and 16% of the injected labelling was recovered in the eggs, 3/4 being free 20E. When 20E was injected during oviposition, it was not converted to the polar products but only to the apolar products. At this time, 76% of the total radiolabel injected accumulated in the egg-batch, principally in the form of free unmetabolized 20E. After injection of [3H]-ecdysone ([3H]-E), the three polar metabolites 1', 2' and 3', probably 20-deoxy homologues of 1, 2 and 3 described above were always produced irrespective of the time of injection. In addition, E was metabolized to 20E and to the apolar conjugates AP1, AP2, and AP3. E, 20E and peak 2' were incorporated into the ovary within the first day after injection. These 3 compounds were found in freshly laid eggs in variable proportions, the quantity of E decreasing with time while 20E and peak 2' increased. At the end of oviposition, ca. 60% of the injected radiolabel had been incorporated into the eggs. Apolar products and polar metabolites accumulating in the body were apparently not used as a source of free hormone for the eggs. Our results with tritiated ecdysteroids confirm our data concerning endogenous ecdysteroids of the eggs of A. hebraeum (Connat et al. 1985). This species, in contrast to 2 other female ticks, Ornithodoros moubata and Boophilus microplus, incorporates free E and 20E instead of ecdysteroid conjugates into its eggs. The role of these free ecdysteroids remains to be elucidated.

Metabolism of [ 3H]ecdysone by isolated tissues of the female ixodid tick Amblyomma hebraeum (Ixodoidea; Ixodidae)

Malpighian tubules, gut, ovaries and carcasses of the adult female tick Amblyomma hebraeum were incubated in vitro in the presence of 2 μM [ 3H]ecdysone. Organs and media were separately extracted after 6, 24 and 48 h incubations and the patterns of ecdysone metabolites were analyzed by HPLC. Esterase-susceptible apolar metabolites similar to the AP2 already described in the soft tick Ornithodoros moubata and thus presumably corresponding to the same conjugates (C-22 esters with fatty acids) were rapidly produced in all tissues investigated. They were mainly found within the organs but they were also released into the medium to some extent. By contrast, less apolar metabolites corresponding to the AP1 esters were mainly found in the media. Malpighian tubules and gut were the most active organs regarding the conversion of ecdysone into 20-hydroxyecdysone (20E). However, only low quantities of 20E were formed, reaching respectively 12.5% and 11.6% of the total metabolites after 48 h incubations. In the carcass and in the ovary the formation of 20E was only a minor pathway (1.7% and 3.1% of the total metabolites after 48 h). In ovaries we observed a massive conversion of ecdysone into 3-epiecdysone, which (as in insects) presumably proceeded through the intermediate formation of 3-dehydroecdysone. These two compounds were identified among the metabolites by CI/D mass spectrometry. The 3-epimer was released into the media, in contrast with the AP2 which were essentially stored within ovaries. Epimerization was also realized to some extent by carcasses, and again the epimer was released into the culture media. The different pathways are compared with those found in other tick species and in insects, and the significance of the various metabolites is discussed.

Effects of octopamine on fluid secretion by isolated salivary glands of a feeding ixodid tick

AbstractOctopamine elicited a dose‐related secretory response by salivary glands isolated from the feeding female tick Amblyomma americanum. Half‐maximal stimulation occurred at about 60 μM. Phentolamine (10 μM) failed to inhibit the octopamine‐mediated response; however, thioridazine (50 μM) inhibited both octopamine (1,000 μM) and dopamine‐stimulated (0.1 μM) secretion. Maximal stimulation by dopamine (1.0 μM) showed no further increase in the rate of secretion after adding octopamine (1,000 or 0.1 μM). Glands responded to octopamine (100 μM) with rates significantly lower than controls following exposure to amphetamine (1,000 μM). Octopamine receptors do not appear to mediate the secretory response, and octopamine may stimulate secretion by releasing catecholamines from presynaptic neurons. These results support the hypothesis that dopamine is the natural transmitter mediating fluid secretion in the feeding tick salivary gland.