Abstract

Our recent study revealed some early molecular and cellular events in which 17β-estradiol (E2) disrupted testis differentiation and resulted in feminization in Xenopus laevis (the African clawed frog), an ideal species for studying reproductive endocrine disruption by estrogenic endocrine disrupting chemicals (EDCs). On this basis, we aimed to develop multiple biomarkers for early detection of testis differentiation disruption by estrogenic EDCs in X. laevis. Tadpoles at stage 45/46 were exposed to four known estrogenic EDCs with different estrogenic activities, including E2, diethylstilbestrol (DES), mestranol (MES) and 4-n-nonyphenol (NP). At stage 53, gonadal morphological and histological changes as well as altered sex-dimorphic gene expression in gonad-mesonephros complexes (GMCs) showed that these estrogenic EDCs disrupted testis differentiation and caused feminization to different degrees. Then we measured transcriptional changes of 48 candidate genes, which are believed to be associated with E2-induced testis differentiation alterations, in GMCs at stage 50. As a result, 19 genes were found to be transcriptionally altered by all test chemicals and proposed as promising biomarkers for early detection of testis differentiation disruption by estrogenic EDCs. Finally, all biomarker responses were integrated as integrated biomarker response (IBR) index to characterize testis differentiation disruption by these estrogenic EDCs in X. laevis. Compared with the methods used in previous studies, the multiple biomarker test using X. laevis at early developmental stages largely shortens the exposure duration, thereby achieving the goal of rapid detection. Certainly, the biomarker test needs further validations in the future study.

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