Identification of transcriptional effects of ethynyl oestradiol in male plaice (Pleuronectes platessa) by suppression subtractive hybridisation and a nylon macroarray Margaret Brown a, * , Ian M. Davies b , Colin F. Moffat b , Craig Robinson b , John Redshaw c , John A. Craft a a Biological and Biomedical Sciences, Glasgow Caledonian University, Cowcaddens Rd., Glasgow, Scotland, UK b Fisheries Research Services, Marine Laboratory, Aberdeen, Scotland, UK c Scottish Environment Protection Agency, Peel Park, East Kilbride, Scotland, UK Abstract Suppression subtractive hybridisation (SSH) was used to generate cDNA libraries repre- senting genes differentially expressed in response to ethynyl oestradiol (EE2) exposure in liver from male plaice (Pleuronectes platessa) previously analysed for vitellogenin (VTG) induction. Characterisation of the cDNA clones identified many as VTG (2 genes) and zona radiata proteins (ZRP) (3 genes), but 40 encoded other proteins, with more than half cryptic. Further analysis identified 85 non-redundant clones suitable for array on nylon membrane. Radiola- belled cDNAs were prepared from hepatic mRNA from EE2 treated plaice (0 and 21 days) and hybridised with the arrayed clones. Analysis of the data showed that 11/17 novel, 21/22 VTG, 13/14 ZRP, 2/2 liver aspartic proteinase (LAP) and 8/10 other mRNAs were up-regu- lated by EE2 exposure. Crown copyright Ó 2004 Published by Elsevier Ltd. All rights reserved. Keywords: Pleuronectes platessa; Suppression subtractive hybridisation; Ethynyl oestradiol; Endocrine disruption; Transcription profiling * Corresponding author. Tel.: +44-141-331-3222; fax: +44-141-331-3208. E-mail address: m.a.brown@gcal.ac.uk (M. Brown). 0141-1136/$ - see front matter. Crown copyright Ó 2004 Published by Elsevier Ltd. All rights reserved. doi:10.1016/j.marenvres.2004.03.045 www.elsevier.com/locate/marenvrev Marine Environmental Research 58 (2004) 559–563 MARINE ENVIRONMENTAL RESEARCH