Expression of steroidogenic enzymes in porcine polycystic ovaries.

Abstract

Abstract:In the present study the expression pattern of the cholesterol side-chain cleavage cytochrome (P450 scc ), 3β-hydroxysteroid dehydrogenase (3β-HSD) and aromatase (P450 arom ) was analyzed in the health and polycystic ovaries of gilts by means of the Western blot and immunohistochemistry. The polycystic status of ovaries was induced by i.m. dexamethasone (DXM) injections on days 7-21 of the estrous cycle. Macroscopic observation of ovaries of DXM-treated gilts revealed the presence of cysts (1-2 cm in diameter, with a mean number of 7.0±1.2 per ovary), a decrease (P<0.05) in number of small follicles (1-3 mm in diameter), as well as the lack of medium-sized follicles (4-6 mm in diameter) and corpora lutea, as compared to the control animals. The expression of P450 scc (P<0.01), 3β-HSD (P<0.05) and P450 arom (P<0.001) proteins in the cysts was higher than in the medium-sized follicles of the control gilts. Moreover, DXM injections resulted also in an enhancement (P<0.05) in the level of P450 scc protein in the walls of small follicles as compared to the control gilts. Following DXM administration the immunoreactivity (IR) of P450 scc in the primordial follicles was lower than in the control group. Comparing to the control gilts, the reaction for this enzyme in DXM-treated animals was observed in secondary follicles, while for 3β-HSD, in primordial, primary, as well as secondary follicles. The immunostaining for P450 scc (theca cells) and P450 arom (granulosa cells) in the small follicles of the DXM-treated gilts were more prominent than those found in the gonads of control animals. However, IR for P450 scc was not found in the granulosa cells of small follicles in the gilts receiving DXM. The intensity of P450 scc and P450 arom labelling was distinctly enhanced in the cysts as compared to the medium follicles of the control animals. Furthermore, in contrary to the medium follicles of the control animals, faint IR for 3β -HSD was found in the granulosa cell layer of cysts. Our data revealed that both the expression of P450 scc , 3β-HSD and P450 arom and localization of these enzymes in polycystic ovaries were different from those, found under physiological conditions. These results suggested that above-mentioned enzymes may, by influencing the ovarian steroid synthesis, play an essential role in the creation and/or course of cystic ovarian disease.

Localization of substance P, calcitonin gene related peptide and galanin in the nerve fibers of porcine cystic ovaries

“…Also, long-term estrogen treatment led to a decrease in the number of both noradrenergic and ERs-expressing ovarian neurons in the caudal mesenteric ganglion of adult gilts [39]. It is worth noting that in the gilts from which cystic ovaries were collected for the present study, the levels of estrogens and androstendione increased in the peripheral blood (unpublished data), and the intraovarian expression of steroidogenic enzymes also changed [26]. It is possible that changes in steroid metabolism in cystic ovaries partly result from DXM, as has been indicated previously [40].…”

“…Follicular structures exceeding 1.0 cm in diameter were classified as cysts [25]. Morphological examination of ovaries particularly focused on the results described previously by Kozłowska et al [26], including on the number of partly luteinized follicular cysts (Figure 1), a decrease in the number of follicles measuring 1-3 mm in diameter, and a lack of follicles measuring 4-6 mm in diameter.…”

Supraphysiological leptin levels shift the profile of steroidogenesis in porcine ovarian follicles toward progesterone and testosterone secretion through increased expressions of CYP11A1 and 17β-HSD: a tissue culture approach

“…By acting on the above-mentioned enzymes to influence ovarian steroid synthesis, leptin may play an essential role in the creation and/or course of cyst formation both in prepubertal and in regularly cycling animals. It has previously been reported that alterations to the concentrations of steroid hormones in the cystic ovaries of swine (Kozłowska et al 2009) are accompanied by changes in the expressions and cellular distributions of CYP11A1, 3b-HSD, 17,20-lyase, 20a-HSD, and/or CYP19. Kozłowska et al (2009) showed that the expression of CYP11A1, -CYP17, -17b-HSD in cysts was higher than that in MFs of control gilt pigs.…”

“…It has previously been reported that alterations to the concentrations of steroid hormones in the cystic ovaries of swine (Kozłowska et al 2009) are accompanied by changes in the expressions and cellular distributions of CYP11A1, 3b-HSD, 17,20-lyase, 20a-HSD, and/or CYP19. Kozłowska et al (2009) showed that the expression of CYP11A1, -CYP17, -17b-HSD in cysts was higher than that in MFs of control gilt pigs. Moreover, an injection of DXM resulted in increased levels of P450scc in the walls of SFs relative to control gilt pigs.…”

“…Sirotkin & Meszarosová (2010) noted an increase in P 4 secretion by porcine granulosa cells after the addition of 1 and 10 ng/ml leptin but observed no effect at a dose of 100 ng/ml. Kozłowska et al (2009) published a study demonstrating the expressions and localization patterns of CYP11A1, 3b-hydroxysteroid dehydrogenase (3b-HSD) and CYP19 in porcine ovaries with a polycystic status induced by dexamethasone (DXM) injections on days 7-21 of the estrous cycle. They found higher expressions of these markers in the follicles from polycystic pigs than in the medium-sized follicles from control pigs.…”