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Title: Steroids and development of the rat Wolffian duct
Author: Marchetti, Nicholas Ian
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2005
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The aims of the studies described in this thesis were threefold. Firstly, to establish an in vitro culture system for the Wolffian ducts; secondly, to elucidate the role of androgens in the development and differentiation of the Wolffian duct from day 19 p.c. onwards, and finally to examine the impact of steroids and steroidogenic compounds on protein expression in the developing Wolffian duct. Immunoexpression of six markers was examined in the freshly isolated Wolffian duct on days 19.5 p.c. through to d0, the day of birth: the androgen receptor (AR), the two oestrogen receptors (ERα and ERβ), the two 5a-reducatase isotypes (types 1 and 2) and a maker of testicular differentiation, smooth muscle α-actin. AR was expressed throughout the duct in both stromal and epithelial cells. ERβ expression followed a similar pattern to AR. ERα expression was localised to the epithelial cells of the efferent ducts. Both 5α-reductase isotypes were found to be immunoexpressed in epithelial cells along the length of the Wolffian ducts. Smooth muscle α-actin was localised to the peri-ductal stromal cell layer immediately surrounding the epithelial cells, and expression proceeded temporally in a cranial-caudal direction. Wolffian ducts were cultured successfully in vitro. The elongation/convolution of the ductal lumen responded to testosterone in a dose-dependent manner. Flutamide, an anti-androgen caused the duct to die within 48 hours of addition in culture. The ducts did not respond to the addition of oestrogens to the culture medium. The efferent ducts were found to be necessary for survival of the ducts in vitro. Following in vitro culture, the same six immunohistochemical markers were examined, and compared to their immunoexpression before culture. No major differences were noted. To elucidate the role of androgens in vivo, developing fetuses were exposed to three compounds in utero: the anti-androgen flutamide, the synthetic oestrogen diethylstilboestrol (DES), and di-n-butyl phthalate (DBP). Again, immunoexpression of the six markers was examined at day 19 p.c. following in utero exposure and compared to controls. Exposure to DES in fetal life caused an increase in immunoexpression of smooth muscle α-actin at the distal end of the Wolffian duct, perhaps indicating premature differentiation of the stromal tissue. Exposure to DBP in vivo reduced the concentration of testosterone in homogenates of fetal rat testes.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available