Retinoic acid affects basement membrane formation of the seminiferous cords in 14-day male rat gonads in vitro

Abstract

The vitamin A derivative retinoic acid (RA) has been previously shown to have teratogenic effects and an ability to modulate cell differentiation in vivo and in vitro. In this study bilateral testicular primordia with the mesonephroi attached were isolated from rat fetuses at 14.5 days of gestation. The gonads were cultured on agar-coated grids in a synthetic medium. RA was added to male rat embryonic gonad cultures at a final concentration of 10 −6 M for 3 h. Two types of controls were prepared: (1) by omitting RA from the culture medium (alcohol controls) and (2) by using plain medium (untreated controls). When applied to gonad cultures RA was found to affect basement membrane development and disturb the general appearance of the tissue. All controls exhibited normal morphology. In order to evaluate the morphological changes observed due to the RA treatment, constituents of the basement membrane, laminin and collagen IV, were localized immunohistochemically at the light microscope level. Basement membrane was also studied at the electron microscope level in control and RA-treated cultures. We propose that one of the effects RA has on rat testicular morphogenesis is the irreversible suppression of seminiferous cord basement membrane formation and the disruption of normal testicular morphogenesis.