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Peroxisome proliferator activated receptors (PPARs) are transcriptional factors belonging to the nuclear receptor superfamily. Three members of the PPAR family have been characterized as PPARα, PPARβ and PPARγ, encoded by separate genes. These isoforms exhibit diverse expression patterns and are activated by different ligands that cause various biological effects. The PPARs are mainly responsible for homeostasis and lipid metabolism. Recently it has been discovered that PPARs play a crucial role in the female reproductive system. PPARγ is expressed in the ovary, endometrium, placenta and fetal membranes. The lack of functional receptors PPARγ or PPARβ results in aberrant placentation and leads to lethality in rodents. PPARβ is abundantly expressed at implantation sites in mice uteri which suggests its role in implantation. Summarizing, in the recent decade studies have revealed new functions of PPARs as the family of receptors that regulate reproductive processes.
We have studied in the porcine endometrium the expression of oxytocin receptor (OTR) mRNA and the effect of progesterone (P₄) on oxytocin/oxytocin receptor (OT/OTR) function concerning intracellular Ca²⁺mobilisation ([Ca²⁺]i), prostaglandin F2α (PGF2α) and E2 (PGE2; PG) secretion. Tissue was taken from cyclic and early pregnant pigs (days 14-16). A higher expression of OTR mRNA (P<0.05) was observed in the endometrium of cyclic than pregnant pigs. The stimulatory (P<0.05) effect of OT (10⁻⁷ M) on [Ca²⁺]i mobilisation was noticed within 15-60 s and 30-60 s in endometrial stromal cells of cyclic and pregnant pigs, respectively. In the presence of P₄ (10⁻⁵ M) basal and OT-stimulated [Ca²⁺]i concentrations decreased in stromal cells during luteolysis and pregnancy. In stromal cells P₄ delayed mobilisation of [Ca²⁺]i in response to OT by 15 s during luteolysis and had no effect during pregnancy. In cyclic and pregnant epithelial cells OT stimulated mobilisation of [Ca²⁺]i in 45 s and 60 s, respectively. Oxytocin increased (P<0.05) PGF2α secretion during luteolysis and pregnancy and PGE2 during luteolysis from endometrial slices. Progesterone did not inhibit this stimulatory effect. During luteolysis OT increased (P<0.05) PGF2α in epithelial and stromal cells and PGE2 secretion in epithelial cells. In the presence of P₄ this effect of OT was reduced only in stromal cyclic cells (6 h culture). The presence of P₄ decreased the effect of OT on [Ca²⁺]i mobilisation only in stromal cells. We found that, in most conditions, P₄ did not inhibit the OT-stimulated secretion of PG in the porcine endometrium.
The aim of the present study was to evaluate the possible direct effects of GnRH, oxytocin (OT) and vasoactive intestinal peptide (VIP) on the release of LH and PRL by dispersed porcine anterior pituitary cells. Pituitary glands were obtained from mature gilts, which were ovariektomized (OVX) one month before slaughter. Gilts randomly assigned to one of the four groups were treated: in Group 1 (n=8) with 1 ml/100 kg b.w. corn oil (placebo); in Group 2 (n=8) and Group 3 (n=8) with estradiol benzoate (EB) at the dose 2.5 mg/100 kg b.w., respectively, 30-36 h and 60-66 h before slaughter; and in Group 4 (n=9) with progesterone (P4) at the dose 120 mg/100 kg b.w. for five consecutive days before slaughter. In gilts of Group 2 and Group 3 treatments with EB have induced the negative and positive feedback in LH secretion, respectively. Isolated anterior pituitary cells (106/well) were cultured in McCoy's 5a medium with horse serum and fetal calf serum for 3 days at 37°C under the atmosphere of 95% air and 5% CO2. Subsequently, the culture plates were rinsed with fresh McCoy's 5A medium and the cells were incubated for 3.5 h at 37°C in the same medium containing one of the following agents: GnRH (100 ng/ml), OT (10-1000 nM) or VIP (1-100 nM). The addition of GnRH to cultured pituitary cells resulted in marked increases in LH release (p<0.001) in all experimental groups. In the presence of OT and VIP we noted significant increases (p<0.001) in LH secretion by pituitary cells derived from gilts representing the positive feedback phase (Group 3). In contrast, OT and VIP were without any effect on LH release in Group 1 (placebo) and Group 2 (the negative feedback). Pituitary cells obtained from OVX gilts primed with P4 produced significantly higher amounts (p<0.001) of LH only after an addition of 100 nM OT. Neuropeptide GnRH did not affect PRL secretion by pituitary cells obtained from gilts of all experimental groups. Oxytocin also failed to alter PRL secretion in Group 1 and Group 2. However, pituitary cells from animals primed with EB 60-66 h before slaughter and P4 produced markedly increased amounts of PRL in the presence of OT. Neuropeptide VIP stimulated PRL release from pituitary cells of OVX gilts primed with EB (Groups 2 and 3) or P4. In contrast, in OVX gilts primed with placebo, VIP was without any effect on PRL secretion. In conclusion, the results of our in vitro studies confirmed the stimulatory effect of GnRH on LH secretion by porcine pituitary cells and also suggest a participation of OT and VIP in modulation of LH and PRL secretion at the pituitary level in a way dependent on hormonal status of animals.
The study was conducted to determine gene expression of short form of leptin receptor (OB-Rs) using real time RT-PCR in distinct tissues of the central nervous system (medial basal hypothalamus, preoptic area, stalk median eminence), pituitary and reproductive tract (corpus luteum, ovarian stroma, endometrium, myometrium, and trophoblast) in pigs during luteal phase of the cycle and early gestation. The expression of OB-Rs mRNA in SME did not differ between analyzed stages of the cycle and pregnancy. In anterior pituitary, transcript levels were almost identical in mid- and late-luteal periods, but significantly decreased on 30-32 day of gestation when compared with day 14-16. In posterior pituitary, significantly higher expression was observed in two periods of pregnancy when compared with two stages of luteal phase. In corpus luteum the lowest expression was observed during days 10-12 of the cycle, whereas markedly higher levels were detected in late-luteal stage and gestation. In ovarian stroma the expression of Ob-Rs mRNA was markedly diminished during days 14-16 of the cycle when compared with days: 10-12 of the cycle and 30-32 of pregnancy. The expression of Ob-Rs mRNA in endometrium and myometrium reached the lowest levels on 30-32 day of pregnancy in comparison with earlier stage, 14-16 day. Summarizing, the expression of the short form of leptin receptor mRNA was found in majority of tested tissues including hypothalamus, pituitary and reproductive tract and their levels fluctuated depending on the phase (mid- and late-luteal) of the cycle and the day of pregnancy (early and late stage of implantation).
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