Assessing gene function in the ruminant placenta.

The placenta provides the means for nutrient transfer from the mother to the fetus, waste transfer from the fetus to the mother, protection of the fetus from the maternal immune system, and is an active endocrine organ. While many placental functions have been defined and investigated, assessing the function of specific genes expressed by the placenta has been problematic, since classical ablation-replacement methods are not feasible with the placenta. The pregnant sheep has been a long-standing animal model for assessing in vivo physiology during pregnancy, since surgical placement of indwelling catheters into both maternal and fetal vasculature has allowed the assessment of placental nutrient transfer and utilization, as well as placental hormone secretion, under unanesthetized-unstressed steady state sampling conditions. However, in ruminants the lack of well-characterized trophoblast cell lines and the inefficiency of creating transgenic pregnancies in ruminants have inhibited our ability to assess specific gene function. Recently, sheep and cattle primary trophoblast cell lines have been reported, and may further our ability to investigate trophoblast function and transcriptional regulation of genes expressed by the placenta. Furthermore, viral infection of the trophoectoderm layer of hatched blastocysts, as a means for placenta-specific transgenesis, holds considerable potential to assess gene function in the ruminant placenta. This approach has been used successfully to "knockdown" gene expression in the developing sheep conceptus, and has the potential for gain-of-function experiments as well. While this technology is still being developed, it may provide an efficient approach to assess specific gene function in the ruminant placenta.

Effect of a CIDR insert and flunixin meglumine, administered at the time of embryo transfer, on pregnancy rate and resynchronization of estrus in beef cattle.

The objectives of this study were to evaluate the effects of flunixin meglumine (FM), an inhibitor of PGF(2alpha) synthesis, and insertion of an intravaginal progesterone-releasing device (CIDR), on pregnancy rates in beef cattle embryo transfer (ET) recipients, and to examine the effect of a CIDR after embryo transfer on the synchrony of the return to estrus in non-pregnant recipients. Cows (n=622) and heifers (n=90) at three locations were assigned randomly to one of four groups in a 2x2 factorial arrangement of treatments with FM administration (500 mg i.m.) 2-12 min prior to ET, and insertion of a CIDR (1.38 g progesterone) immediately following ET as main effects. Fresh or frozen embryos (Stage=4 or 5; Grade=1 or 2) were transferred on Days 6-9 of the estrous cycle and CIDR devices were removed 13 days after ET. Recipients at Location 2 only were observed for signs of return to estrus. Recipients that returned to estrus at Location 2 were either bred by AI or received an embryo 7 days after estrus. Following the initial ET, there was an FMxlocation interaction on pregnancy rate (P<0.01; Location 1, 89% versus 57%; Location 2, 69% versus 64%; Location 3, 64% versus 67% for FM versus no FM, respectively). Pregnancy rates of embryo recipients were not affected by CIDR administration (P>0.05; 65% with CIDR, 70% without CIDR), however, the timing of the return to estrus was more synchronous (P<0.01) for recipients given a CIDR. Pregnancy rate of recipients bred following a return to estrus did not differ between cows receiving or not receiving a CIDR for resynchronization (P>0.13). Effects of FM on pregnancy rate were location dependent and CIDR insertion at ET improved synchrony of the return to estrus.

Effect of P.G. 600 on the timing of ovulation in gilts treated with altrenogest.

We previously reported that ovulation rate, but not pregnancy rate or litter size at d 30 after mating, was enhanced by treatment with P.G. 600 (400 IU of PMSG and 200 IU of hCG, Intervet America, Inc., Millsboro, DE) in gilts fed the orally active progestin, altrenogest (Matrix, Intervet America, Inc.) to synchronize estrus. We hypothesized that in addition to increasing ovulation rate, P.G. 600 may have altered the timing of ovulation. Therefore, mating gilts 12 and 24 h after first detection of estrus, as is common in the swine industry, may not have been the optimal breeding regimen, and as a consequence, pregnancy rate and litter size were not altered. The objective of the present study was to determine the effect of P.G. 600 on the timing of ovulation in gilts treated with altrenogest. Randomly cycling, crossbred gilts (5.5 mo old, 117 kg BW, and 14.7 mm of backfat) were fed a diet containing altrenogest (15 mg/d) for 18 d. Twenty-four hours after altrenogest withdrawal, gilts received i.m. injections of P.G. 600 (n = 25) or saline (n = 25). Gilts were checked for estrus at 8-h intervals. After first detection of estrus, transrectal ultrasonography was performed at 8-h intervals to determine the time of ovulation. Gilts were killed 9 to 11 d after the onset of estrus to determine ovulation rate. All gilts displayed estrus by 7 d after treatment with P.G. 600 or saline. Compared with saline, P.G. 600 increased (P = 0.07) ovulation rate (14.8 vs. 17.5, respectively; SE = 1.1). The intervals from injection to estrus (110.9 vs. 98.4; SE = 2.7 h; P < 0.01) and injection to ovulation (141.9 vs. 128.6; SE = 3.2 h; P < 0.01) were greater in gilts treated with saline than in gilts treated with P.G. 600. Duration of estrus (54.4 vs. 53.7; SE = 2.5 h), the estrus-to-ovulation interval (30.2 vs. 31.7; SE = 2.2 h), and the time of ovulation as a percentage of estrus duration (55.8 vs. 57.5; SE = 3.0%) did not differ for the P.G. 600 and saline-injected gilts, respectively. In summary, P.G. 600 advanced the onset of estrus and ovulation following termination of altrenogest treatment and increased ovulation rate; however, treatment of gilts with P.G. 600 had no effect on the timing of ovulation relative to the onset of estrus.