Caenorhabditis elegans as an environmental monitor using DNA microarray analysis.
In order to assist in the identification of possible endocrine disrupting chemicals (EDC) in groundwater, we are developing Caenorhabolitis elegans as a high throughput bioassay system in which responses to EDC may be detected by gene expression using DNA microarray analysis. As a first step we examined gene expression patterns and vitellogenin responses of this organism to vertebrate steroids, in liquid culture. Western blotting showed the expected number and size of vitellogenin translation products after estrogen exposure. At 10(-9) M, vitellogenin decreased, but at 10(-7) and 10(-5), vitellogenin was increased. Testosterone (10(-5) M) increased the synthesis of vitellogenin, but progesterone-treated cultures (10(-5) M) had less vitellogenin. Using DNA microarray analysis, we examined the pattern of gene expression after progesterone (10(-5), 10(-7), and 10(-9) M), estrogen (10(-5) M), and testosterone (10(-9) M) exposure, with special attention to the traditional biomarker genes used in environmental studies [vitellogenin, cytochrome P450 (CYP), glutathione s-transferase (GST), metallothionein (MT), and heat shock proteins (HSP)]. GST and P450 genes were affected by estrogen (10(-5) M) and progesterone (10(-5) and 10(-7) M) treatments. For vitellogenin genes, estrogen treatment (10(-5) M) caused overexpression of the vit-2 and vit-6 genes (2.68 and 3.25 times, respectively). After progesterone treatment (10(-7) M), the vit-5 and vit-6 were down-regulated and vit-1 up-regulated (3.59-fold). Concentrations of testosterone and progesterone at 10(-9) M did not influence the expression of the vit, CYP, or GST genes. Although the analysis is incomplete, and low doses and combinations of EDC need to be tested, these preliminary results indicate C. elegans may be a useful laboratory and field model for screening EDC.
Custodia
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N
Department of Biology, Boston University, Massachusetts 02215, USA.
Won
S J
SJ
Novillo
A
A
Wieland
M
M
Li
C
C
Callard
I P
IP
D000818:Animals; D015153:Blotting, Western; D017173:Caenorhabditis elegans / Q000187:drug effects* / Q000235:genetics; D002784:Cholesterol / Q000633:toxicity; D004527:Egg Proteins / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004784:Environmental Monitoring / Q000379:methods*; D005260:Female; D020869:Gene Expression Profiling; D020411:Oligonucleotide Array Sequence Analysis; D011374:Progesterone / Q000633:toxicity; D012333:RNA, Messenger / Q000096:biosynthesis / Q000302:isolation & purification; D013256:Steroids / Q000633:toxicity*; D013739:Testosterone / Q000633:toxicity; D014158:Transcription, Genetic; D014819:Vitellogenins / Q000032:analysis / Q000096:biosynthesis; D014874:Water Pollutants, Chemical / Q000633:toxicity*
D004527:Egg Proteins; D012333:RNA, Messenger; D013256:Steroids; D014819:Vitellogenins; D014874:Water Pollutants, Chemical; D013739:Testosterone; D011374:Progesterone; D002784:Cholesterol