Messenger ribonucleic acid for insulin-like growth factors-I and -II, insulin-like growth factor-binding protein-2, gonadotropin receptors, and steroidogenic enzymes in porcine follicles

Abstract

Several aspects of growth, development, and steroidogenesis in ovarian follicles are controlled by the interaction of insulin-like growth factors (IGF-I and IGF-II) and gonadotropins (LH and FSH). The objective of this study was to determine the developmental stage and location at which mRNA for IGF, IGF-binding protein (IGFBP)-2, gonadotropin receptors, and steroidogenic enzyme genes are expressed within porcine follicles. Ovaries from pigs on Days 10 (n = 3; midluteal phase) and 19 (n = 3; preovulatory phase) of the estrous cycle were collected, frozen, and sectioned. The mRNA for IGF-I, IGF-II, IGFBP-2, LH receptor, FSH receptor, cytochrome P450 side-chain cleavage (P450scc), 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), 17 alpha-hydroxylase (P45017 alpha), and aromatase (P450arom) were detected by in situ hybridization, and relative amounts were determined by image analysis. Expression of mRNA for IGF-I and IGF-II was cell specific (granulosa and theca interna cells, respectively) and increased with follicular size. The IGFBP-2 mRNA was expressed in greater amounts for theca interna cells than for granulosa cells. The pattern of expression for LH and FSH receptors diverged as follicles grew. For LH receptor, mRNA was detected in the theca interna cells of small antral follicles and in both the theca interna and granulosa cells of large antral follicles. Amount of LH receptor mRNA increased with follicular diameter. For FSH receptor, mRNA increased as follicles grew to a small antral size. However, FSH receptor mRNA disappeared in large antral follicles on Day 19 of the estrous cycle. Steroidogenic enzyme mRNA for 3 beta-HSD (theca interna cells), P450scc (theca interna cells), P45017 alpha (theca interna cells), and P450arom (granulosa cells) increased with follicular diameter. In summary, coordinated expression of IGF, IGFBP-2, gonadotropin receptors, and steroidogenic enzyme mRNA throughout different stages of follicular development was observed. Changes in mRNA suggested increased IGF-I and -II action and a switch from FSH to LH dependence in porcine preovulatory follicles.