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

A Conesa

A Del Valle

A Henriksen

A Loukas

AL Williamson

Alex Loukas

B Ewing

B Zhan

Bennett J. D. Datu

CT Murphy

D Longbottom

Eng K. Ong

G Braun

J Bethony

J Tort

J Wang

JH McKerrow

JM Hawdon

JM Moser

John Dalton

JW Golden

L Diatchenko

M Mitreva

M Moyle

M Smid

ML Blaxter

ML Rhoads

MS Chan

MS Duffy

MW Pfaffl

ND Rawlings

NR de Silva

OA Asojo

P Geldhof

P Syntichaki

Peter O'Donoghue

PJ Hotez

PS Albert

RB Gasser

RM Maizels

Robin B. Gasser

Russell McInnes

S Draghici

S Hashmi

S Lustigman

S Trivedi

Shivashankar H. Nagaraj

Shoba Ranganathan

X Huang

PLoS Neglected Tropical Diseases

PubMed

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

Bennett J D Datu

Robin B Gasser

Shivashankar H Nagaraj

Eng K Ong

Directory of Open Access Journals

Transcriptional changes in the hookworm, Ancylostoma caninum, during the transition from a free-living to a parasitic larva.

Crossref

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

Datu, Bennett J. D

Gasser, Robin B

Nagaraj, Shivashankar

Ong, Eng K

O⿿Donoghue, Peter

McInnes, Russell L

Ranganathan, Shoba

Loukas, Alex

Macquarie University ResearchOnline

Datu, BJD

Gasser, RB

Nagaraj, SH

Ong, EK

O'Donoghue, P

McInnes, R

Ranganathan, S

Loukas, A

University of Melbourne Institutional Repository

Background\ud
\ud
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.\ud
\ud
Methodology/Principal Findings\ud
\ud
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.\ud
\ud
Conclusions/Significance\ud
\ud
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

Datu, Bennett J.D.

Gasser, Robin B.

Nagaraj, Shivashankar H.

Ong, Eng K.

O'Donoghue, Peter

McInnes, Russell

Name not available

Transcriptional changes in the hookworm, ancylostoma caninum, during the transition from a free-living to a parasitic larva

 by culturing L3 in serum-containing medium. 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. 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

Datu Bennett J. D.

Gasser Robin B.

Nagaraj Shivashankar H.

Ong Eng K.

O'Donoghue Peter

McInnes Russell

Ranganathan Shoba

Loukas Alex

Public Library of Science (PLOS)

, during the Transition from a Free-Living to a Parasitic Larva

Datu, Bennett J. D.

University of Queensland eSpace

Transcriptional changes in the hookworm, Ancylostoma caninum, during the transition from a free-living to a parasitic larva

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

Gasser, Robin

ResearchOnline at James Cook University

English

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

Datu, Bennett

Hiriyur Nagaraj, Shivashankar

Ong, Eng

Queensland University of Technology ePrints Archive

A developmentally regulated hyaluronidase of Haemonchus contortus.

A developmentally regulated metalloprotease secreted by host-stimulated Ancylostoma caninum third-stage infective larvae is a member of the astacin family of proteases.

A hookworm glycoprotein that inhibits neutrophil function is a ligand of the integrin CD11b/CD18.

A new mathematical model for relative quantification in realtime RT-PCR.

Albumin and a dialyzable serum factor stimulate feeding in vitro by third-stage larvae of the canine hookworm Ancylostoma caninum.

Ancylostoma caninum MTP-1, an astacin-like metalloprotease secreted by infective hookworm larvae, is involved in tissue migration.

Ancylostoma secreted protein 2: cloning and characterization of a second member of a family of nematode secreted proteins from Ancylostoma caninum.

Base-calling of automated sequencer traces using phred. I. Accuracy assessment.

Base-calling of automated sequencer traces using phred. II. Error probabilities.

Blast2GO: a universal tool for annotation, visualization and analysis in functional genomics research.

Cathepsin B homologue at the interface between a parasitic nematode and its intermediate host.

Characterisation of the two most abundant genes in the Haemonchus contortus expressed sequence tag dataset.

Cleavage of hemoglobin by hookworm cathepsin D aspartic proteases and its potential contribution to host specificity.

Cloning and characterization of Ancylostoma-secreted protein. A novel protein associated with the transition to parasitism by infective hookworm larvae.

Cloning of a cysteine protease required for the molting of Onchocerca volvulus third stage larvae.

Developmental alterations in sensory neuroanatomy of the Caenorhabditis elegans dauer larva.

Effect of vaccination with a recombinant fusion protein encoding an astacinlike metalloprotease (MTP-1) secreted by host-stimulated Ancylostoma caninum thirdstage infective larvae.

Evaluation of endogenous reference genes for realtime PCR quantification of gene expression in Ancylostoma caninum.

Gene expression profiles associated with the transition to parasitism in Ancylostoma caninum larvae.

Genes expressed in Brugia malayi infective third stage larvae.

Genes that act downstream of DAF-16 to influence the lifespan of Caenorhabditis elegans.

Genetic markers in ribosomal DNA for hookworm identification.

Global analysis of dauer gene expression in Caenorhabditis elegans.

Global functional profiling of gene expression.

GO-Mapper: functional analysis of gene expression data using the expression level as a score to evaluate Gene Ontology terms.

Hookworm aspartic protease, Na-APR-2, cleaves human hemoglobin and serum proteins in a host-specific fashion.

Hookworm vaccines: past, present, and future.

Hookworm: developmental biology of the infectious process.

Immunological crossreactivity between a cloned antigen of Onchocerca volvulus and a component of the retinal pigment epithelium.

Invasion of skin by Schistosoma cercariae.

Investigating hookworm genomes by comparative analysis of two Ancylostoma species.

Isolation and molecular cloning of a secreted hookworm platelet inhibitor from adult Ancylostoma caninum.

Long-term storage of hookworm infective larvae in buffered saline solution maintains larval responsiveness to host signals.

Loukas A

Madan A

Major venom allergen of yellow jackets, Ves v 5: structural characterization of a pathogenesis-related protein superfamily.

MEROPS: the peptidase database.

Molecular characterisation of the Ancylostoma-secreted protein family from the adult stage of Ancylostoma caninum.

Molecular cloning and characterisation of a putative aspartate proteinase associated with a gut membrane protein complex from adult Haemonchus contortus.

Proteinases and associated genes of parasitic helminths.

Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR.

Serum-stimulated feeding in vitro by third-stage infective larvae of the canine hookworm Ancylostoma caninum.

Soiltransmitted helminth infections: ascariasis, trichuriasis, and hookworm.

Soiltransmitted helminth infections: updating the global picture.

Specific aspartyl and calpain proteases are required for neurodegeneration in C.

Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries.

The Caenorhabditis elegans cathepsin Z-like cysteine protease, Ce-CPZ-1, has a multifunctional role during the worms’ development.

The Caenorhabditis elegans dauer larva: developmental effects of pheromone, food, and temperature.

The evaluation of potential global morbidity attributable to intestinal nematode infections.

X-ray structure of Na-ASP-2, a pathogenesis-related-1 protein from the nematode parasite, Necator americanus, and a vaccine antigen for human hookworm infection.