Schistosoma mansoni: DNA microarray gene expression profiling during the miracidium-to-mother sporocyst transformation.
For the human blood fluke, Schistosoma mansoni, the developmental period that constitutes the transition from miracidium to sporocyst within the molluscan host involves major alterations in morphology and physiology. Although the genetic basis for this transformation process is not well understood, it is likely to be accompanied by changes in gene expression. In an effort to reveal genes involved in this process, we performed a DNA microarray analysis of expressed mRNAs between miracidial and 4 d old in vitro-cultured mother sporocyst stages of S. mansoni. Fluorescently labeled, dsDNA targets were synthesized from miracidia and sporocyst total RNA and hybridized to oligonucleotide DNA microarrays containing 7335 S. mansoni sequences. Fluorescence intensity ratios were statistically compared between five biologically replicated experiments to identify particular transcripts that displayed stage-associated expression within miracidial and sporocyst mRNA populations. A total of 361 sequences showed stage-associated expression in miracidia, while 273 probes displayed sporocyst-associated expression. Differentially expressed mRNAs were annotated with gene ontology terminology based on BLAST homology using high throughput gene ontology functional annotation toolkit (HT-GO-FAT) and clustered using the GOblet GO browser software. A subset of genes displaying stage-associated expression by microarray analyses was verified utilizing real-time quantitative PCR. The use of DNA microarrays for the profiling of gene expression in early-developing S. mansoni larvae provides a starting point for expanding our understanding of the genes that may be involved in the establishment of parasitism and maintenance of infection in these important life cycle stages.
Vermeire
Jon J
JJ
Department of Pathobiological Sciences, University of Wisconsin-Madison, 2115 Observatory Drive, Madison, WI 53706, USA.
Taft
Andrew S
AS
Hoffmann
Karl F
KF
Fitzpatrick
Jennifer M
JM
Yoshino
Timothy P
TP
D000818:Animals; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D015801:Helminth Proteins / Q000235:genetics / Q000378:metabolism*; D007814:Larva / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D033661:Oocysts / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012550:Schistosoma mansoni / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D014158:Transcription, Genetic
D015801:Helminth Proteins; D012333:RNA, Messenger