Insulin-like growth factor-I (IGF-I) stimulates the development of cultured rat pre-antral follicles

The aim of the present study is to investigate the effect of insulin-like growth factor (IGF-I) on the development of cultured rat pre-antral follicles. Pre-antral follicles with a diameter between 140 and 160 microm are mechanically isolated from ovaries of 10-day-old rats and cultured in groups for 6 days in FSH and insulin-containing serum-free medium in the absence or presence of IGF-I at concentrations of 1, 10, and 100 ng/ml, respectively. DNA content of the follicles before and after culture is measured to evaluate whether a possible growth is due to proliferation of follicular cells and the ultrastructure of the cultured follicles is studied with transmission electron microscopy to evaluate the quality of follicles cultured under different conditions. Furthermore, reverse transcriptase polymerase chain reaction (RT-PCR) is used to assess the gene expression of IGF-I and type I IGF receptor in pre-antral follicles. When follicles were cultured in medium containing IGF-I at a concentration of 1 and 10 ng/ml, the increase in follicle diameter after a 6-day culture differs significantly from that in the absence of IGF-I. The ultrastructure of follicles cultured in IGF-I-containing medium is better sustained when compared to that of follicles cultured in the absence of IGF-I. Especially, theca cells, which are undergoing degeneration under controlled culture condition, demonstrate normal cellular ultrastructure with some characteristics of steroid-secreting cells in the presence of IGF-I. Moreover, in oocytes of follicles cultured in the presence of IGF-I, cortical granules are observed distributed along the ooplasma membrane whereas cortical granules are hardly present in follicles cultured without IGF-I. With RT-PCR, the presence of the mRNA of IGF-I and type I IGF receptor is demonstrated in both the somatic follicular cells and the oocytes. Taken together, these results suggest that IGF-I, in the presence of follicle-stimulating hormone (FSH), enhances rat pre-antral follicle development in vitro and supports the morphology of cultured pre-antral follicles. The stimulatory effect of exogenous IGF-I on the development of pre-antral follicle together with the gene expression of both IGF-I and type I IGF receptor in pre-antral follicles suggests the involvement of IGF-I in early folliculogenesis and its actions are likely mediated by the putative type I IGF receptor, via both endocrine and paracrine/autocrine pathways.