Funding priorities in animal reproduction at the United States Department of Agriculture's Cooperative State Research, Education, and Extension Service.

The National Research Initiative (NRI) Competitive Grants Program is the U.S. Department of Agriculture's major competitive grants program and is administered by the Cooperative State Research, Education, and Extension Service (CSREES). Since its inception in 1991, the NRI has funded competitive grants in the discipline of animal reproduction. Previously, this program provided funding for a broad range of projects encompassing almost every subdiscipline in reproductive biology of farm animals, including aquatic species important to the aquaculture industry. During fiscal year 2004, the NRI Animal Reproduction Program narrowed the focus of funding priorities to the topics of infertility, basic mechanisms regulating fertility, cryopreservation of gametes, reducing the postpartum interval to conception, and sterilization methods or development of monosex populations. In response to a directive to further narrow the focus of funding priorities for fiscal year 2005 and beyond, CSREES conducted a Stakeholder Workshop on Funding Priorities in Animal Reproduction at the 37th Annual Meeting of the Society for the Study of Reproduction in Vancouver, Canada. More than 75 stakeholder scientists from a cross section of federal, public, and private institutions from across the United States participated in the workshop and provided recommendations to CSREES for future NRI-funding priorities in Animal Reproduction. The recommendations provided by stakeholders included continuing efforts to focus funding priorities into fewer high-impact areas relevant to animal agriculture and aquaculture. Recommendations also included movement back toward subdisciplines of animal reproduction that cut across all applicable species. The three funding priorities that consistently emerged as recommendations from the workshop participants were 1) gonadal function and production of gametes, 2) pituitary-hypothalamic function, and 3) embryo and conceptus development, including interaction between the conceptus and uterus. These funding priorities were considered when preparing the fiscal year 2006 NRI Request for Applications.

Endocrine secretion of prostaglandin F2alpha in cyclic gilts is decreased by intrauterine administration of exogenous oxytocin.

When administered systemically, oxytocin (OT) stimulates secretion of uterine prostaglandin F2alpha (PGF2alpha) in swine, but the role of endometrially-derived OT in control of PGF2alpha release is not clear. This study determined the effect of exogenous OT, administered into the uterine lumen of intact cyclic gilts, on PGF2alpha secretion during late diestrus. Intrauterine infusion of 40USP units OT (in 30 ml 0.9% saline) was performed for 30 min (1 ml/min) into each uterine horn between 7:00 and 9:00 h on days 10, 12, 14 and 16 after estrus. Beginning 20 min before infusion, samples of jugular venous blood were drawn at 5-10-min intervals for 140 min for quantification of 13,14-dihydro-15-keto-PGF2alpha (PGFM), the major stable metabolite of PGF2alpha. Progesterone was analyzed in samples collected 0, 60 and 120 min after initiation of OT infusion. Treatment with OT did not alter plasma concentrations of PGFM on days 10 or 12 but decreased (P<0.001) PGFM concentrations for 40 min after onset of infusion on day 16. Concentrations of PGFM also were reduced in the pre-treatment samples on day 14 (P=0.05) and day 16 (P<0.001) in OT-infused gilts. Plasma progesterone declined (P<0.01) between days 10 and 16 in control-infused gilts but did not decline until after day 14 (P<0.001) in gilts infused with OT. These results indicate that when OT is administered into the uterine lumen of pigs during late diestrus, it has an anti-luteolytic effect to reduce endocrine secretion of PGF2alpha and delay the decline in progesterone that occurs during luteolysis.

Effect of ovarian steroids on basal and oxytocin-induced prostaglandin F2alpha secretion from pig endometrial cells.

These studies were undertaken to determine how treatment with 100 nM progesterone and/or 10 nM oestradiol-17beta acutely (3 h; Experiment 1) or chronically (72 h; Experiments 2-4) influenced basal and oxytocin (OT)-stimulated prostaglandin (PG) F(2alpha) secretion, in enriched cultures of pig endometrial luminal epithelial, glandular epithelial and stromal cells obtained on Day 16 (Experiments 1, 2 and 4) or Day 12 (Experiment 3) after oestrus. In Experiment 1, acute treatment with progesterone stimulated PGF(2alpha) secretion from each cell type on Day 16, whereas acute oestradiol treatment inhibited the stimulatory action of progesterone on PGF(2alpha) secretion only in glandular epithelial cells. In Experiment 2, OT stimulated phospholipase (PL) C activity in luminal epithelial cells on Day 16 only in the presence of chronic oestradiol treatment. For glandular epithelial cells on Day 16, OT stimulated PLC activity only in the presence of chronic treatment with steroid. In stromal cells on Day 16, OT stimulated PLC activity in the absence of steroids and the response to OT was further enhanced by oestradiol. In the absence of chronic treatment with steroid, OT did not stimulate PGF(2alpha) secretion from luminal epithelial cells, but oestradiol induced a response to OT. For glandular epithelial cells, OT-induced PGF(2alpha) secretion was not altered by steroids, whereas the stimulatory response to OT was inhibited by oestradiol or progesterone in stromal cells. For endometrial cells obtained on Day 12 after oestrus in Experiment 3, OT only stimulated PGF(2alpha) release from glandular epithelial and stromal cells. For luminal epithelial cells obtained on Day 16 after oestrus and cultured under polarizing conditions in Experiment 4, secretion of PGF(2alpha) occurred preferentially from the basolateral surface and was stimulated by OT more from the basolateral surface than from the apical surface. Oxytocin-induced PGF(2alpha) secretion from the apical surface was enhanced by chronic treatment with oestradiol, whereas that from the basolateral surface was enhanced by chronic treatment with progesterone. In summary, oestradiol enhanced OT-induced PGF(2alpha) secretion from the apical surface of luminal epithelial cells and reduced the response of stromal cells to OT, actions that may contribute to the reorientation of PGF(2alpha) from endocrine secretion (i.e. towards the uterine vasculature) to exocrine secretion (i.e. towards the uterine lumen) during pregnancy recognition in pigs.

The role of essential trace elements in embryonic and fetal development in livestock.

This review addresses the concept that essential trace minerals play a vital role in many enzymatic and metabolic pathways that are critical for conceptus development during pregnancy in livestock species. The conceptus relies entirely on the maternal system for a sufficient supply of trace minerals and other nutrients needed for normal development. If this supply is inadequate, growth and/or health of the conceptus can be affected adversely, and many of these effects carry over into the neonatal period. Information, accumulated in our laboratory and presented herein, indicates that zinc, copper and manganese are among the trace minerals that have the greatest impact on reproduction. For example, levels of zinc, copper and manganese were several fold greater in the conceptus than in other reproductive tissues, indicating that the conceptus preferentially accumulates these minerals, an action that may be important for conceptus development, growth and survival. Moreover, some recent results indicate that increasing the biological availability of zinc, copper and manganese, by attachment to short peptide chains (i.e., proteinated trace minerals) can enhance reproductive performance of swine. Mineral concentrations in conceptuses from female pigs consuming proteinated trace minerals were greater than those from females that consumed only inorganic mineral salts. Elucidating the mechanisms whereby conceptus development and survival are enhanced by essential trace minerals may lead to development of specific feeding programs to increase the number and health of offspring at parturition, thereby allowing for further improvements in production efficiency in animal agriculture.

The effects of chronic ethanol consumption during early pregnancy on conceptus health and uterine function in pigs.

BACKGROUND: Maternal ethanol consumption impairs fetal health, but it is unclear if this occurs through direct actions on the conceptus or indirectly through effects on the uterus. The objective of this study was to determine if chronic ethanol consumption in swine would impair early embryonic and fetal health either through direct effects on the conceptus or indirect effects on the endometrium. METHODS: Four experiments evaluated the effects of chronic ethanol consumption during early pregnancy. Female pigs were fed either 350 ml of 95% ethanol or an isocaloric amount of dextrose at 10 to 14-hr intervals beginning on day 10 after pubertal estrus and continuing until ovariohysterectomy 11 to 35 days after mating. At the second estrus, pigs were mated to a fertile boar that did not consume alcohol. RESULTS: In experiment 1, ethanol consumption increased (p < 0.01) blood alcohol concentrations that peaked 2-3 hr after feeding. In experiment 2, ethanol was detectable in uterine flushings 2 hr after feeding on day 11 of pregnancy and was highly correlated (r = 0.989, p < 0.001) with blood alcohol concentration. In experiment 3, ethanol consumption did not affect endometrial phospholipase C activity on days 11 and 16 of pregnancy but decreased (p < 0.05) basal endometrial prostaglandin F(2alpha) production on day 16. However, ethanol consumption did not decrease the number of conceptuses on day 11 or conceptus DNA content on days 11 or 16. In experiment 4, ethanol consumption decreased (p < 0.05) fetal survival rate to 58% versus 85% in dextrose-fed controls on day 35 of pregnancy. For viable conceptuses, ethanol consumption reduced (p < 0.01) fetal weight, fetal crown-rump length, placental weight and volume of placental (chorio-allantoic + amniotic) fluid. CONCLUSION: These results indicate that chronic ethanol consumption may impair conceptus health directly or indirectly through actions upon the endometrium. Thus, the pig may be a valuable experimental model for studies on the effects of maternal alcohol consumption on conceptus development.

Morphometric analysis of the uterine endometrium of swine on days 12 and 16 postestrus.

The uterine endometrium of swine is comprised of luminal epithelial, glandular epithelial, and stromal cells that secrete the luteolysin, prostaglandin F(2alpha) (PGF(2alpha)), during late diestrus. However, which of these cells contribute the most to luteolytic PGF(2alpha) secretion is unknown because the cellular composition of the endometrium has not been quantified. Therefore, this study quantified the cellular composition of the endometrium on days 12 and 16 postestrus by histologic and morphometric analyses. On day 12, the endometrium consisted predominantly of stromal cells (47% of total cell quantity) and glandular epithelial cells (37%), whereas luminal epithelial cells represented only 16% of the total of the three cell types. The number of glandular epithelial cells tended to increase (P < 0.10) between days 12 and 16, such that they comprised 45% of the endometrium by day 16, while the number of stromal and luminal cells did not change and accounted for 45% and 10% of the cells, respectively. Luminal epithelial cells had a 58% greater cross-sectional area (P < 0.001) than glandular epithelial cells, whereas glandular epithelial cells had a 22% greater area (P < 0.001) than stromal cells. Glandular epithelial cells decreased (P < 0.001) in cross-sectional area between days 12 and 16, whereas the area of luminal epithelial and stromal cells remained unchanged. These results indicate that the porcine endometrium is comprised predominantly of stromal and glandular epithelial cells that are likely to contribute substantially to endometrial PGF(2alpha) secretion during luteolysis. The contribution of glandular epithelium to luteolytic PGF(2alpha) secretion probably increases during diestrus as the number of these cells increases.

Angiotensin II increases intracellular calcium concentration in pig endometrial stromal cells through type 1 angiotensin receptors, but does not stimulate phospholipase C activity or prostaglandin F2alpha secretion.

Although the presence of endometrial receptors for angiotensin (Ang) II has been demonstrated, a specific function for AngII in the uterus has not been identified. Cytosolic free Ca2+ concentration [Ca2+]i, phospholipase C (PLC) activity and prostaglandin (PG) F2alpha secretion in response to AngII and oxytocin (OT) were measured in pig endometrial stromal cells collected 16 days after oestrus. Treatment with 100 nM OT or AngII increased (P<0.001) [Ca2+]i in stromal cells similarly (720 +/- 34 v. 690 +/- 33 pM, respectively). Subsequent administration of OT or AngII to the same cells induced smaller [Ca2+]i increases (25% or 35% of the initial responses, respectively) that occurred only if the second exposure to the same agent took place at least 5 min after the first. When administered sequentially, OT and AngII each induced a full response within 1 min of the previous treatment, regardless of which peptide was applied first. Whereas OT increased PLC activity and PGF2alpha secretion in stromal cells (P<0.01), AngII did not increase either PLC activity or PGF2alpha secretion. Type I AngII (AT1) receptors were present on stromal cells, whereas AT2 receptors were absent. Therefore, the effect of AngII in stromal cells was mediated via AT1 receptors. That AngII increased [Ca2+]i in stromal cells, but did not increase PLC or PGF2alpha secretion, indicates that either AngII releases a pool of Ca2+ through a mechanism that is not mediated by PLC and is not involved in PGF2alpha secretion or that a mechanism for PGF2alpha production other than one involving Ca2+ may exist.