Development of the ovary and ontongeny of mRNA and protein for P450 aromatase (arom) and estrogen receptors (ER) alpha and beta during early fetal life in cattle.
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Estradiol-17beta is the predominant steroid produced during early stages of ovarian development in ruminants and steroid hormones have been hypothesized to regulate ovigerous cord formation, germ cell meiosis and ovarian vascular development. Therefore, the objective was to determine the presence and localization of mRNA and protein encoding cytochrome P450 aromatase (P450arom), and estrogen receptors alpha (ERalpha) and beta (ERbeta) during ovarian development in fetuses of cattle on days 35, 45, 60, 75, 90 and 105 after breeding (n=4/age) using in situ hybridization and immunohistochemistry. No ovarian tissue was found in the day 35 fetuses, but was found in all later ages studied. There appeared to be little organization of specific structures in ovaries on days 45 and 60, although germ cells could be identified. Evidence of the beginning of ovigerous cord formation was found on day 60. By day 75 of gestation, the ovigerous cords were more extensive and mesonephric-derived cell streams were detectable. By day 90 (and still present at day 105), both ovigerous cords and cell streams/rete tubules were definitive structures of the developing ovaries. Ovaries appeared to develop in "lobular" segments around the periphery of the ovary. Some lobes appeared to be at slightly different developmental stages, as assessed by the extent or definition of ovigerous cord formation. The localization of mRNAs for P450arom, ERalpha and ERbeta were closely associated with protein content. At days 45 and 60, mRNA and protein of P450arom and ERbeta were located throughout ovaries with signal in medulla being denser than in the cortex. P450arom mRNA or protein was punctate, but not evident in germ cells. From day 75, P450arom was increasingly becoming localized to cell streams or clusters of cells (rete tubules) in the medulla, and by days 90 and 105 of gestation, was more definitively localized to cell streams and/or rete tubules. Similar to P450arom, ERbeta mRNA and protein were observed in cells in the medulla, and also in germ cells, pre-granulosa cells and some surface epithelial cells. ERalpha mRNA and protein were predominately in the surface epithelium in ovaries of all ages with fainter signal for ERalpha protein also being observed in pre-granulosa and stromal cells including the cell streams/rete tubules. ERalpha protein was also detected in a few germ cells at days 90 and 105 of gestation. Thus, in cattle, estradiol-17beta has the potential to regulate, in an autocrine/paracrine manner, a number of different cell types during ovarian development.