title: Characterisation of genes transcriptionally upregulated in the liver of sand goby (Pomatoschistus minutus) by 17α-ethinyloestradiol: identification of distinct vitellogenin and zona radiata protein transcripts.
abstract: The sand goby (Pomatoschistus minutus), is a marine and estuarine teleost that is used in environmental, reproductive and behavioural studies of oestrogenic endocrine disruption. The xeno-oestrogen, 17α-ethinyloestradiol (EE2), induces expression of egg proteins vitellogenin (VTG) and zona radiata protein (ZRP) in male fish and impairs reproduction. Multiple forms of VTG and ZRP genes are found in other teleost species, yet the characteristics of VTG and ZRP in the sand goby are unknown. In this investigation, Suppressive Subtractive Hybridization was used to isolate cDNA fragments from liver, identified as belonging to 11 distinct sand goby genes, suggesting that these genes are transcriptionally upregulated by EE2. Assembly of these fragments revealed three VTG genes which shared homology with VTG classes A, B and C in other fish and two ZRP genes sharing homology with ZRP classes Ba and Bb. RTqPCR of RNA from the sand goby liver was used to show that these VTGs and ZRPs were present in low levels in control males and high levels in mature females. Exposure of males to a concentration of 11 ngL(-1) EE2 caused a significant increase in all VTG and ZRP transcript levels. The identification of these egg protein transcripts and the development of validated assays for their quantification will facilitate future work with this useful model species.
authors.affiliation: School of Health and Life Sciences, Glasgow Caledonian University, Cowcaddens Road, Glasgow G4 0BA, United Kingdom. joseph.humble@gcu.ac.uk
authors.forename: Joseph L
authors.initials: JL
authors.lastname: Humble
authors.forename: Elaine
authors.initials: E
authors.lastname: Hands
authors.forename: Minna
authors.initials: M
authors.lastname: Saaristo
authors.forename: Kai
authors.initials: K
authors.lastname: Lindström
authors.forename: Kari K
authors.initials: KK
authors.lastname: Lehtonen
authors.forename: Oihane
authors.initials: O
authors.lastname: Diaz de Cerio
authors.forename: Ibon
authors.initials: I
authors.lastname: Cancio
authors.forename: Gemma
authors.initials: G
authors.lastname: Wilson
authors.forename: John A
authors.initials: JA
authors.lastname: Craft
mesh_terms: D000818:Animals; D004527:Egg Proteins / Q000235:genetics* / Q000378:metabolism; D052244:Endocrine Disruptors / Q000633:toxicity*; D004997:Ethinyl Estradiol / Q000633:toxicity*; D005260:Female; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D008297:Male; D010473:Perciformes; D014819:Vitellogenins / Q000235:genetics* / Q000378:metabolism; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D004527:Egg Proteins; D052244:Endocrine Disruptors; D029941:Fish Proteins; D014819:Vitellogenins; D014874:Water Pollutants, Chemical; C423083:zona radiata protein, fish; D004997:Ethinyl Estradiol