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Testosterone selectively increases primary follicles in ovarian cortex grafted onto embryonic chick membranes: relevance to polycystic ovaries

A Qureshi, Cellular and Molecular Medicine, St George's University of London, London, United Kingdom
S Nussey, Cellular and Molecular Medicine, St George's University of London, London, United Kingdom
G Bano, Cellular and Molecular Medicine, St George's University of London, London, United Kingdom
P Musonda, Community Health Sciences, St George's University of London, London, United Kingdom
S Whitehead, Division of Basic Medical Sciences, St George's University of London, London, United Kingdom
H Mason, Clinical Developmental Sciences, St George's University of London, London, United Kingdom

Correspondence: Asjid Qureshi, Email: asjidqureshi{at}hotmail.com

Abstract

Histological studies demonstrated that polycystic ovaries contain increased numbers of preantral follicles with a specific increase in primary follicles. Polycystic ovary syndrome is associated with hyperandrogenism and pre- and postnatal androgenisation of primates increases the pool of growing follicles producing changes resembling polycystic ovaries. In vitro studies could test the hypothesis that androgens alter early folliculogenesis, but conventional culture techniques for small follicles are generally unsuitable in non-rodent species. Our objective was to develop and use a method to investigate the effects of testosterone on early folliculogenesis. We adapted an in ovo technique in which lamb cortical ovarian fragments were grafted onto the chorioallantoic membrane of fertilised chick eggs. Optimal experimental conditions for vascularisation and survival of tissue were determined and the model then used to investigate the effects of testosterone on follicle growth. Eggs were inoculated with testosterone at the time of implantation of the ovarian tissue which was retrieved 5 days later. Tissue was sectioned and follicles staged and counted. There was no wholesale initiation of primordial follicle growth over the 5-day in ovo culture. Importantly, the proportion of primordial, primary and secondary follicles remained similar to those in unimplanted tissue. Testosterone increased the number of primary follicles by 50% compared to controls, an effect which was largely due to a reduction in atresia. In conclusion, incubation of ovarian cortex with testosterone reproduces the changes in early folliculogenesis reported in histological studies of polycystic ovaries.