The effect of chronic steroid treatment of ovariectomized ewes on LH-RH priming effect, exogenous LH-RH induced LH output and pituitary LH content

Abstract

The objective of this study was to determine whether gonadotrophin-releasing hormone (GnRH), oxytocin (OT) and vasoactive intestinal polypeptide (VIP) modulate β-endorphin-like immunoreactivity (β-END-LI) secretion by dispersed anterior pituitary cells of pigs and in vivo priming with steroid hormones, estradiol benzoate (EB) and progesterone (P4), influences the cell reactivity to peptide hormones tested. Additionally, the aim of this research was to examine the involvement of cyclic nucleotides (cAMP and cGMP) in transduction of signals induced by GnRH, OT and VIP in porcine pituitary cells. Pituitaries were collected from ovariectomized (OVX) gilts that were divided into four experimental groups. Animals of group 1 (OVX) received 1 ml corn oil (placebo)/100 kg body weight (b.w.), group 2 (OVX+EB I) and group 3 (OVX+EB II) were treated with EB at the dose 2.5 mg/100 kg b.w., 30–36 and 60–66 h before slaughter, respectively. Animals of group 4 (OVX+P4) were injected with P4 at the dose 120 mg/100 kg b.w. for 5 subsequent days before slaughter. Anterior pituitaries were dispersed with trypsin and then pituitary cells were cultured (106 per well) in McCoy’s 5A medium containing horse serum (10%) and fetal calf serum (2.5%) for 3 days at 37 °C under an atmosphere of 95% air and 5% CO2. Subsequently, plates were rinsed with fresh McCoy’s 5A medium and pituitary cells were treated with one of the following agents: GnRH (100 ng/ml), OT (10−6 M) or VIP (10−7 M) and incubated for 3.5 h at 37 °C.GnRH did not affect β-END-LI secretion by pituitary cells of OVX (group 1) and OVX+P4 (group 4) gilts. When the pituitary cells were incubated in the presence of OT and VIP, significant increases were observed. After priming of OVX gilts with EB, 30–36 h before slaughter (group 2), we noted a significant increase in β-END-LI release from pituitary cells only in the presence of VIP. Pituitary cells from gilts treated with EB, 60–66 h before slaughter (group 3), produced markedly elevated amounts of β-END-LI after GnRH, OT or VIP addition.GnRH markedly stimulated cGMP release from cultured pituitary cells in all experimental groups and significantly increased cAMP production by the cells from OVX, OVX+EB II and OVX+P4 animals. The addition of OT enhanced both cAMP and cGMP output in all experimental groups of pigs. VIP stimulated cAMP release from pituitary cells derived from OVX, OVX+EB I and OVX+EB II animals. cGMP output was markedly elevated under the influence of VIP from pituitary cells of OVX, OVX+EB II and OVX+P4 gilts.In conclusion, our results suggest that GnRH, OT and VIP can modulate β-endorphin release from porcine pituitary cells and imply the involvement of cAMP and cGMP in transduction of signals induced by studied peptides in the cells.