Transcriptional Changes in the Hookworm, Ancylostoma caninum, during the Transition from a Free-Living to a Parasitic Larva

Autor: Shoba Ranganathan, Bennett J. D. Datu, Peter J. O'Donoghue, Robin B. Gasser, Russell L. McInnes, Eng K. Ong, Alex Loukas, Shivashankar H. Nagaraj
Jazyk: angličtina
Rok vydání: 2008
Předmět:
Zdroj: PLoS Neglected Tropical Diseases
PLoS Neglected Tropical Diseases, Vol 2, Iss 1, p e130 (2008)
ISSN: 1935-2735
1935-2727
Popis: Background Third-stage larvae (L3) of the canine hookworm, Ancylostoma caninum, undergo arrested development preceding transmission to a host. Many of the mRNAs up-regulated at this stage are likely to encode proteins that facilitate the transition from a free-living to a parasitic larva. The initial phase of mammalian host invasion by A. caninum L3 (herein termed “activation”) can be mimicked in vitro by culturing L3 in serum-containing medium. Methodology/Principal Findings The mRNAs differentially transcribed between activated and non-activated L3 were identified by suppression subtractive hybridisation (SSH). The analysis of these mRNAs on a custom oligonucleotide microarray printed with the SSH expressed sequence tags (ESTs) and publicly available A. caninum ESTs (non-subtracted) yielded 602 differentially expressed mRNAs, of which the most highly represented sequences encoded members of the pathogenesis-related protein (PRP) superfamily and proteases. Comparison of these A. caninum mRNAs with those of Caenorhabditis elegans larvae exiting from developmental (dauer) arrest demonstrated unexpectedly large differences in gene ontology profiles. C. elegans dauer exiting L3 up-regulated expression of mostly intracellular molecules involved in growth and development. Such mRNAs are virtually absent from activated hookworm larvae, and instead are over-represented by mRNAs encoding extracellular proteins with putative roles in host-parasite interactions. Conclusions/Significance Although this should not invalidate C. elegans dauer exit as a model for hookworm activation, it highlights the limitations of this free-living nematode as a model organism for the transition of nematode larvae from a free-living to a parasitic state.
Author Summary Hookworms are soil-transmitted nematodes that parasitize hundreds of millions of people in developing countries. Here we describe the genes expressed when hookworm larvae make the transition from a developmentally arrested free-living form to a tissue-penetrating parasitic stage. Ancylostoma caninum can be “tricked” into thinking it has penetrated host skin by incubating free-living larvae in host serum – this is called “activation”. To comprehensively identify genes involved in activation, we used suppressive subtractive hybridization to clone genes that were up- or down-regulated in activated larvae, with a particular focus on up-regulated genes. The subtracted genes, as well as randomly sequenced (non-subtracted) genes from public databases were then printed on a microarray to further explore differential expression. We compared predicted gene functions between activated hookworms and the free-living nematode, Caenorhabditis elegans, exiting developmental arrest (dauer), and found enormous differences in the types of genes expressed. Genes encoding secreted proteins involved in parasitism were over-represented in activated hookworms whereas genes involved in growth and development dominated in C. elegans exiting dauer. Our data implies that C. elegans dauer exit is not a reliable model for exit from developmental arrest of hookworm larvae. Many of these genes likely play critical roles in host-parasite interactions, and are therefore worthy of pursuit for vaccine and drug development.
Databáze: OpenAIRE
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