Morphological and transcriptomic effects of endocrine modulators on the gonadal differentiation of chicken embryos: the case of tributyltin (TBT)

Chicken eggs (Gallus gallus domesticus) were incubated, after 24 h of incubation either non-injected (control, C), or injected with the solvent sesame oil (Oleum Sesame Raffinatum, solvent control, SC), and the substances Tributyltin (TBT, 10pg TBT-SN/g egg) or 17alpha-Methyltestosteron (MT, 30 pg/g egg). Animals were decapitated on breeding day 19, 2 days before anticipated hatching. Right and left gonads of female and male chicken were accurately separated from adhesing tissues, stored in RNA lysis buffer (Promega) at -80°C until until RNA isolation with the SV Total RNA Isolation System Kit according to manual 048 (Promega). Sequencing was performed on Illumina's Genome Analyzer IIx, and subsequent base calling was carried out by Illumina's GAPipeline. Our study represents the first detailed analysis of whole chicken gonad transcriptomes, with biologic replicates, generated by SuperSAGE technology. Overall design: SuperSAGE analysis of substance-induced whole-gemome expression in 19-days-bred chicken gonads