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Type 1 NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase (PGDH) is the key enzyme for metabolism of active primary prostaglandins to inactive forms in gestational tissues. The present study examined the activity and immunolocalization of PGDH in the ovine placenta, fetal membranes and uterus over the latter half of pregnancy, and its potential regulation by oestradiol. Placenta, fetal membranes and myometrium were collected from sheep with known single insemination dates on days 70, 100 and 135 of gestation and in active labour demonstrated by electromyographic activity. In addition, chronically catheterized fetuses were infused with oestradiol (100 microgram kg(-1) per 24 h) (n = 5) or saline vehicle into the fetus from day 120 to day 125. PGDH activity measured in placental extracts remained constant from day 70 to day 135 of gestation, and then significantly (P < 0.05) increased by 300% in active labour. Immunoreactive PGDH was localized in the placentome at all stages and was present predominantly in the fetal component of the placentome in uninucleate, but not in binucleate, trophoblast cells. Similarly, in the fetal membranes PGDH immuno-reactivity was present in the uninucleate trophoblast but not in the binucleate cells of the chorion. PGDH immunostaining was also present in the endometrial luminal epithelium, in the smooth muscle of the myometrium, and the glandular epithelium of the cervix. Infusion of oestradiol into the fetal circulation from day 120 to day 125 of gestation had no effect on placental PGDH activity. Immunohistochemistry was used to localize oestrogen receptor alpha in intrauterine tissues to investigate further the failure of oestradiol to increase PGDH activity. Immunoreactive oestrogen receptor alpha was not present in the fetal component of the placenta, although it was expressed in adjacent maternal-derived cells. It is concluded that (1) PGDH activity increases in late gestation; (2) PGDH is expressed in uninucleate trophoblast cells in the ovine placenta and fetal membranes, and also in the maternal endometrial epithelium and stroma, myometrium and cervix; (3) oestrogen receptor alpha is not expressed in fetal cells in the placenta or fetal membranes; and (4) the increase in PGDH activity is not regulated by oestradiol administered to the fetus.

Placental growth and development is crucial for successful pregnancy. The aim of this study was to characterize the activity and localization of the matrix metalloproteinase 2 (MMP-2) and MMP-9, which are capable of degrading basement membrane collagen (predominantly collagen type IV), and their endogenous tissue inhibitor of matrix metalloproteinases (TIMPs), in amniotic fluid and in the developing ovine placenta. Cell deletion by apoptosis during placental development was also examined. Zymography with gelatin as substrate indicated that MMP-2 (72 kDa gelatinase A; predominantly latent form) was present in increasing amounts in amniotic fluid from day 70 of gestation to labour (days 140-145), and MMP-9 (92 kDa gelatinase B; predominantly latent form) was detectable from day 125 to labour; there was no increase in MMP-2 or -9 in labour. A broad range of TIMPs was detected in amniotic fluid; the molecular masses corresponded to TIMP-1, -2 and -3. Immunohistochemical techniques localized MMP-2, MMP-9 and TIMP-3 in the sheep placenta, predominantly in the trophoblast layer in uninucleate, but not binucleate, cells. However, MMP-2 and -9 activated proteins in placental homogenates were low throughout pregnancy. Apoptosis was identified by morphological criteria and also by TdT-mediated dUTP nick end labelling. Apoptosis was present in discrete regions in the placenta, predominantly in trophoblast cells near the tips and the basal regions of the fetomaternal interdigitations. During pregnancy the sheep placenta becomes more complex and the area of the fetomaternal interface increases. MMP-2 and -9 are likely to be involved in breaking down basement membranes to allow cell migration during this process. It is suggested that digestion of supporting extracellular matrix may trigger apoptosis and in some way increase the branching pattern in the villi.

An experiment was performed in 20 Merino ewes in which ovarian venous blood was collected by venepuncture at surgery and at two stages of the oestrous cycle. The ovarian venous concentrations of inhibin, oestradiol-17 beta and progesterone were determined. The results demonstrate that during the luteal phase of the oestrous cycle the ovarian venous blood draining an ovary containing luteal tissue contains significantly more inhibin bioactivity than ovarian venous blood from an ovary not containing luteal tissue. During the follicular phase the concentration of inhibin bioactivity in ovarian venous blood was reduced compared with the luteal phase. From this data we conclude that the sheep corpus luteum secretes inhibin bioactivity into the ovarian venous blood.

Abstract: Much research has gone into developing a sustainable management tool for haemonchosis, namely the FAMACHA© system, which has been validated by numerous studies worldwide. However, the financial implications on production of implementing this system have not been fully investigated in sheep previously. A trial was conducted on a farm comprising a flock of approximately 300 Mutton Merinos on which the FAMACHA© system was in use. Seventy five maiden and multiparous ewes were randomly allocated to the following three trial groups: (i) FAMACHA© in which ewes judged to be in categories 4 or 5 were dosed with levamisole HCl 2.5% (Nemasol NF, Intervet); (ii) Conventionally dosed in which all ewes were dosed every six weeks with levamisole HCl 2.5% (Nemasol NF, Intervet); (iii) Suppressively dosed in which all ewes were injected every six weeks with moxydectin 1%m/v (Cydectin, Bayer AH). There were no significant differences in body weight gains, FAMACHA© and body condition scores amongst the three treatment regimens for ewes that did not conceive. For those that became pregnant during the trial there was a significant difference when analysed by calendar date but not when analysed by reproductive stage. There were also no significant differences between trial groups in lamb birth weight and, after slow growers had been sold, the average daily gain of the lambs born to ewes in the trial. The cost of anthelmintics was considerably lower for the FAMACHA© group. In conclusion there were no significant differences in selected production parameters when using the FAMACHA© system as opposed to other methods of anthelmintic use in a Mutton Merino flock in a semi-intensive farming system and that the gestation status should be taken into account when measuring these parameters in future studies. [Copyright &y& Elsevier]

Objectives: We studied collagen plugging of the fetoscopic access site in an in vivo fetal lamb model for fetoscopic surgery and possible role for matrix metalloproteinase (MMP)-2 and -9 and tissue inhibitors (TIMPs).Methods: Eight ewes had fetoscopic balloon occlusion of the trachea as an experimental treatment for congenital diaphragmatic hernia between days 88 and 99 of gestation (term 145 days) with sampling of amniotic, allantoic, and tracheal fluid. Nonoperated cotwins were used as controls. The fetoscopy port was closed using a collagen plug. Ten days (range 9–12) later, fluids were sampled and plug sites collected for histologic analysis. Activity of MMP-2 (72 kDa, gelatinase A) and MMP-9 (92 kDa, gelatinase B) was determined in the fluids by zymography and secretion of TIMPs (27–30 kDa; TIMP-1, glycosylated TIMP-3 and TIMP-4, 24 kDa; unglycosylated TIMP-3, 21 kDa; TIMP-2) by reverse zymography and quantified by densitometric analysis.Results: No pregnancy was complicated by amniorhexis or preterm labor. At cesarean, normal volumes of amniotic and allantoic fluid were present in all cases. Histology of the plug sites revealed good integration of the collagen plug without complete restoration of membrane integrity. MMP-2, MMP-9, and TIMPs were detected in all fluids. In the operated animals, significantly (P < .05) higher activity of MMP-9 was found in amniotic fluid, with lower concentrations of TIMPs in allantoic fluid (P < .01). Tracheal occlusion was associated with a significant (P < .02) increase in both MMP-2 and -9 in tracheal fluid.Conclusion: Collagen plugging of the fetoscopic access port sites in sheep resulted in functionally effective sealing of the fetal membranes. Changes in MMP-2, MMP-9, and TIMPs suggest an active remodeling of both the fetal lung and the fetal membranes. [Copyright &y& Elsevier]