Data from: All-trans retinoic acid disrupts development in ex vivo cultured fetal rat testes. I: altered seminiferous cord maturation and testicular cell fate

Exposure to excess retinoic acid disrupts the development of the mammalian testicular seminiferous cord. However, the molecular events surrounding retinoic acid-driven loss of cord structure have not previously been examined. To investigate the mechanisms associated with this adverse developmental effect, fetal rat testes were isolated on gestational day 15, after testis determination and the initiation of cord development, and cultured in media containing all-trans retinoic acid (ATRA; 10-8 to 10-6 M) or vehicle for 3 days. ATRA exposure resulted in a concentration-dependent decrease in the number of seminiferous cords per testis section and number of germ cells, assessed by histopathology and immunohistochemistry. Following 1 day of culture, genome-wide expression profiling by microarray demonstrated that ATRA exposure altered biological processes related to retinoid metabolism and gonadal sex determination. Real-time RT-PCR analysis confirmed that ATRA enhanced the expression of the key ovarian development gene Wnt4 and the anti-testis gene Nr0b1 in a concentration-dependent manner. After 3 days of culture, ATRA-treated testes contained both immunohistochemically DMRT1-positive and FOXL2-positive somatic cells, providing evidence of disrupted testicular cell fate maintenance following ATRA exposure. We conclude that exogenous retinoic acid disrupts seminiferous cord development in ex vivo cultured fetal rat testes, resulting in a reduction in seminiferous cord number, and interferes with maintenance of somatic cell fate by enhancing expression of factors that promote ovarian development.Supplemental Tables S1-S7Supplemental tables pertaining to microarray analysis of gestation day 15 rat testes cultured for 24 h in media containing 10^-6 M ATRA or DMSO vehicle, including gene lists and pertinent results of Gene Set Enrichment Analysis and Ingenuity Pathway Analysis2018-06-21_I-ATRA_supplemental-tables.xlsx2018-06-21_I-ATRA_supplemental-figuresSupplemental Figures S1-S