Abstract
Vinclozolin is a pesticide with antiandrogenic activity as an endocrine disruptor compound. Its effects upon the progression of primordial follicles were assessed in cultures of mouse fetal ovaries from the onset of meiotic differentiation of germ cells (13.5 days post coitum) and from both in vivo exposed mice and in vitro exposed ovaries. Exposure of ovaries to vinclozolin—at in vitro dosages ranging from 10 to 200 μM and in 3D ex vivo culture following in vivo exposure to 50 mg/kg bw/day—showed delays in meiocyte differentiation and in follicle growth, even at the lowest in vitro dose exposure. Immunofluorescent analysis showed the presence of the proteins MSY2 and NOBOX in the primary follicles but no difference in the level of protein signals or in the number of follicles in relation to treatment. However, assessing the cytological differentiation of germ cells by detecting the synaptonemal complex protein SYCP3, the exposure to vinclozolin delayed meiotic differentiation from both in vitro- and in vivo-exposed ovaries. These effects were concomitant with changes in the energy metabolism, detected as a relative increase of glycolytic metabolism in live-cell metabolic assays in exposed ovaries.
Highlights
Vinclozolin is a pesticide with antiandrogenic activity as an endocrine disruptor compound
The modifications made to the initial culture protocol did not affect the established progression of cell differentiation and folliculogenesis in mouse ovaries in the developmental window from 13.5 dpc followed by 17 days of culture (Fig. 1)
The modifications of the culture procedures allowed us to perform functional analyses of the processes from 3D in vitro cultures and evaluation of the effects of potential reproductive toxicants, such as the endocrine disruptor vinclozolin (VZN), delivered in vitro during the culture process or by the culture of ovaries previously exposed in vivo
Summary
Vinclozolin is a pesticide with antiandrogenic activity as an endocrine disruptor compound. Its effects upon the progression of primordial follicles were assessed in cultures of mouse fetal ovaries from the onset of meiotic differentiation of germ cells (13.5 days post coitum) and from both in vivo exposed mice and in vitro exposed ovaries. Assessing the cytological differentiation of germ cells by detecting the synaptonemal complex protein SYCP3, the exposure to vinclozolin delayed meiotic differentiation from both in vitro- and in vivo-exposed ovaries These effects were concomitant with changes in the energy metabolism, detected as a relative increase of glycolytic metabolism in live-cell metabolic assays in exposed ovaries. Between 10.5 and 11.5 days post coitum (dpc), PGCs colonize the genital ridges They divide mitotically to increase the number of progenitor germ cells, initiating gonadal sex differentiation at about 12.5 dpc[5]. While the incidence of reproductive disorders in females caused by exposure to EDCs, especially during fetal life, is much higher than initially estimated, it has scarcely been assessed in either mice or humans[16,17]
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