Factors affecting follicle and oocyte development in cattle

• Main Author: Ralph, John Hunter
• Published: University of Edinburgh 1997
• Subjects: 636.089
• Online Access: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.660890

The mechanisms governing development of mammalian oocytes are not well understood. Isolation and in vitro growth of immature cattle follicles will enable determination of the factors affecting bovine follicular development, have potential applications in assisted reproduction and provide a suitable model for studying human infertility. Intercommunication of the oocyte and somatic cells is necessary for normal oocyte and follicle development. Studies using systems where oocyte-somatic cell communication is preserved allows an accurate assessment of the factors affecting follicular development. The aims of this project were to examine early follicle and oocyte development in cattle and determine whether the bovine oocyte plays a role in follicular development. A non-enzymatic isolation procedure was developed which allowed intact bovine follicles to be isolated. On the basis of follicle size, these could be divided into 3 distinct stages: large preantral, large preantral/early antral and antral follicles. A culture technique was devised which supported in vitro follicle and oocyte development, the key elements of which were: volume of medium (0.25 ml/follicle), serum and insulin minimal number of medium changes and a substrate of collagen. The effect of FSH on preantral to early antral follicles in culture was examined. Initial experiments on large preantral/early antral follicle growth found that all FSH doses stimulated an increase in follicle diameter. The dose of FSH was important as low levels did not stimulate proliferation or affect oocyte size whilst high levels reduced proliferation, inhibited oocyte growth and reduced oocyte quality. Oocyte localised granulosa cell proliferation was observed in some follicles only when a healthy oocyte was present, demonstrating the importance of oocyte-somatic cell communication in granulosa cell proliferation and differentiation. The intensity of oocyte localised proliferation was reduced at high FSH doses, confirming its dose dependent inhibitory effect on follicular development. FSH stimulated the growth of large preantral/early antral and antral follicles but not oocyte growth in any of the stages. The increase in size was due to an increase in intercellular spacing and, as antral cavities were neither maintained or formed during culture, this may be analogous to antrum development. FSH maintained granulosa cell proliferation in all follicle size classes. No detectable effect of FSH on preantral follicles were found, therefore the effect of FSH depends on the stage of follicle examined.