Modulating peripheral gonadotrophin levels affects follicular expression of mRNAs encoding insulin-like growth factor binding proteins in sheep

Evidence suggests that the insulin-like growth factor binding proteins (IGFBPs) are involved in modulating the role that IGF-I and -II play in regulating follicular growth and development in sheep. However, little information exists as to the role that key peripheral factors play in regulating the expression of IGFBP components within the follicle. The present study investigated the regulatory effects of FSH and LH on gene expression for IGFBP-2 to -6 in ovine follicles, using bovine follicular fluid (bFF) and gonadotrophin-releasing hormone antagonist (GnRHa) model systems to perturb endogenous gonadotrophin secretion. Gene expression studies were carried out using in situ hybridisation with sheep-specific ribonucleotide probes. Treatment of ewes with bFF had few consistent effects on IGFBP-3 and -4 mRNA expression in follicles, whereas bFF treatment resulted in significant decreases in IGFBP-2 and IGFBP-6 mRNA expression in only medium follicles 60 h after onset of bFF, and in follicles 12 h after the onset of bFF, respectively. Treatment of ewes with GnRHa resulted in transient significantly increased IGFBP-2 mRNA in healthy follicles in comparison to early atretic or atretic follicles 12 h post-GnRHa (P < 0.05), in IGFBP-3 mRNA in early atretic and atretic follicles 36 h post-GnRHa (P < 0.001), and IGFBP-5 mRNA in follicles 12 h post-GnRHa (P < 0.001). In contrast, GnRHa treatment resulted in significant decreases in IGFBP-4 (P < 0.001) and IGFBP-6 (P < 0.01) mRNA expression in large follicles 36 h post-GnRHa, and atretic follicles by 60 h post-GnRHa, respectively. These data highlight that FSH and LH are involved, at least in part, in mediating the proliferative and differentiative changes in intrafollicular IGFBP levels that are observed during follicular growth and atresia in the sheep.