title: Phosphoinositide-3-OH-kinase inhibitor LY294002 prevents activation of Ancylostoma caninum and Ancylostoma ceylanicum third-stage infective larvae.
abstract: The developmentally arrested hookworm infective larva resumes development only after encountering specific host-mediated cues during invasion. These cues activate a signaling pathway that culminates in the resumption of development. In Ancylostoma caninum, activation is characterised by the resumption of feeding and the release of excretory/secretory products required for infection. The dauer stage of the free-living nematode Caenorhabditis elegans is a developmentally arrested stage analogous to the hookworm infective larva. Dauer larvae exit developmental arrest in response to environmental cues that indicate favorable conditions for reproduction and growth. Because of the similarity between dauer recovery and activation, exit from dauer provides a model for hookworm larval activation. An insulin-signaling pathway has been implicated in controlling exit from developmental arrest in both C. elegans dauers and A. caninum larvae. To further investigate the role of insulin signaling in hookworm larval activation, the phosphatidylinositol-3-OH kinase inhibitor LY294002 was tested for its effect on in vitro activation using the resumption of feeding as a marker for activation. LY294002 prevented feeding in A. caninum infective larvae stimulated with host serum filtrate and a glutathione-analogue, the muscarinic agonist arecoline, or the cell permeable cGMP-analogue 8-bromo-cGMP. Similar results were seen with the congeneric hookworm Ancylostoma ceylanicum. These data suggest that an insulin-signaling pathway mediates activation in hookworm larvae, as in C. elegans, and that the phosphatidylinositol-3-OH kinase inhibitor acts downstream of the cGMP and muscarinic signaling steps in the pathway. In A. caninum, LY294002 had no effect on the release of excretory/secretory products associated with activation, suggesting that the secretory pathway diverges from the activation pathway upstream of the phosphatidylinositol-3-OH kinase step. These results provide additional support for the insulin-signaling pathway as the primarily pathway for activation to parasitism in hookworm larvae.
authors.affiliation: Department of Microbiology and Tropical Medicine, The George Washington University Medical Center, 725 Ross Hall, 2300 Eye Street North West, Washington DC 20037, USA.
authors.forename: Andrea
authors.initials: A
authors.lastname: Brand
authors.forename: John M
authors.initials: JM
authors.lastname: Hawdon
mesh_terms: D000722:Ancylostoma / Q000187:drug effects* / Q000254:growth & development; D000818:Animals; D001115:Arecoline / Q000378:metabolism / Q000494:pharmacology; D015153:Blotting, Western / Q000379:methods; D002867:Chromones / Q000494:pharmacology*; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D007328:Insulin / Q000378:metabolism; D007814:Larva / Q000187:drug effects; D009025:Morpholines / Q000494:pharmacology*; D018721:Muscarinic Agonists / Q000378:metabolism / Q000494:pharmacology; D000081082:Phosphoinositide-3 Kinase Inhibitors; D015398:Signal Transduction / Q000502:physiology
chemical_list: D002867:Chromones; D004791:Enzyme Inhibitors; D007328:Insulin; D009025:Morpholines; D018721:Muscarinic Agonists; D000081082:Phosphoinositide-3 Kinase Inhibitors; C016276:8-bromocyclic GMP; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D001115:Arecoline; D017869:Cyclic GMP-Dependent Protein Kinases; D006152:Cyclic GMP