Follicle development in the mouse ovary has been studied using two experimental approaches: firstly, a novel culture system was designed to investigate the metabolism of individual follicles for the first time, and secondly, chimaeric mice and molecular biology techniques were employed to study follicle morphogenesis and the developmental relationships between the different ovarian cell types. Physiological studies of follicle metabolism have been limited by the absence of a suitable culture system which is capable of supporting normal follicle growth and maturation. A novel culture system was developed to provide a physiological model for studies of metabolism by individual mouse ovarian follicles. This system supports the growth of individual primary mouse follicles to Graafian stages <i>in vitro</i> over a period of 5 days and is the first model demonstrating apparently normal ovulation of <i>in vitro</i>-grown follicles in response to LH. Preantral follicles of c.180 microns diameter were micro-dissected from the ovaries of 4 week old mice using fine needles and cultured individually in 20 μl of medium under mineral oil in V-shaped wells of a microtitre plate for a period of 6 days. Medium was supplemented with 1IU/ml hFSH and 5% serum from hypogonadal (<i>hpg/hpg</i>) mice. After every 24 hours of culture, follicles were washed and transferred to wells containing 20 μl of fresh medium: samples of 5-10 μl were then taken from the previous well for the analysis of metabolites. During growth and ovulation in culture, follicles and oocytes were morphologically indistinguishable from those during culture, indicating that premature luteinization does not occur in this system. Approximately 60% of dissected follicles reach preovulatory sizes during culture, which is considerably greater than the maturation rate <i>in vivo</i>, indicating that atresia is not a pre-programmed event. Experiments demonstrated that FSH-stimulation is required for full antral development and LH-induced ovulation, confirming that FSH confers LH-responsiveness to the follicle.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:641764 |
| Date | January 1993 |
| Creators | Boland, Nicola I. |
| Publisher | University of Edinburgh |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://hdl.handle.net/1842/20104 |
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