Reproduction in Domestic Animals - Facultad de Ciencias Veterinarias

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16 t h International Congress on Animal Reproduction 86 Poster Abstracts P175 Expression of VEGF in Corpora Lutea of progestagen synchronized-sheep Sanchez, MA 1 *; Garcia-Fernandez, RA 1 ; Letelier, C 2-3 ; Sanchez, B 1 ; Garcia- Palencia, P 1 ; Gonzalez-Bulnes, A 2 ; Flores, JM 1 1Faculty of Veterinary Medicine, UCM, Spain; 2 Animal Reproduction Dpt, INIA, Spain; 3 Faculty of Veterinary Medicine, UACh, Chile Introduction Vascular endothelial growth factor (VEGF) is expressed in blood vessels in thecal derived compartiments and granulosa derived-parenchymal lobules in corpora lutea (CL) 1,2 but there are some discrepancies about its expression in luteal cells 2,3,4 in different species. Though, our aim was to study VEGF expression in different compartiments of the CL in progestagen-induced sheep and elucidate its potential relation with different synchronization treatments. Methods Ovaries from progestagen synchronized (n= 30) and natural cycling (n= 30) Manchega ewes were collected on days 9, 11, 13, 15, 17 and 21 post mating, routinely processed in 4% paraformaldehyde and paraffin-embeded. Later, 41 CL of progestagen synchronized and 48 CL of natural cycling ewes were studied using conventional immunohistochemical techniques with a monoclonal anti VEGF. In every CL were considered 4 different compartiments to evaluate VEGF expression: CL peripheral blood vessels, thecal derived compartiments, granulosa derived-parenchymal lobules and luteal cells. Results VEGF was positive in capillaries and small and medium arteries in the different compartiments studied as well as in luteal cells, but with differences in the intensity of immunostaining among different CL. However, regarding treatments, only on day 9 and in the peripheral blood vessels and granulosa derived-parenchymal lobules capillaries was possible to see a significant increase (p
16 t h International Congress on Animal Reproduction Poster Abstracts 87 P178 Echo-Doppler evaluation of umbilical blood flow during pregnancy in sheep Suarez, G 1 *; Panarace, M 1 *; Cané, L 1 ; Garnil, C 1 ; Medina, M 1 1GOYAIKE S.A.A.C.I. y F., Biotechnology Area. Carmen de Areco CC37. CP 6725. Buenos Aires. Argentina. Introduction Placental blood flow and vascular development are essential components of normal placental function and are critical to fetal growth and development. Umbilical Artery Doppler is an ultrasound technique that allows one to gauge how much resistance fetal blood encounters during its passage through the placenta. Ideally, fetal blood should encounter very little resistance; with certain placental abnormalities, increased resistance to flow may limit oxygenation transfer to fetal blood. Therefore, blood flow in umbilical arteries can be used to monitor placental development and fetal hemodynamics and use these findings for assessing the condition of the fetus. Objective Based on the above mentioned considerations, and considering that most of the studies of umbilical arteries in sheep has been done in instrumented and/or anesthetized fetuses, we aimed to characterize, noninvasively, the Doppler flow velocity waveform in umbilical arteries of ewes with apparently normal pregnancies. Materials and methods Fifteen multiparous, nonlactating Merino breed ewes with singleton pregnancies achieved after natural mating were examined weekly from 4 to 20 weeks of gestation. Ultrasonography (Toshiba Nemio 20, Tokyo, Japan) was performed using two different convex transducers, a 5-10 MHz (transrectal) from 4 to 8 weeks of gestation and thereafter using a 3-6 MHz (transabdominal). Three resistance indices were calculated: S/D ratio, Resistance index (RI) = (S-D)/S, and Pulsatility Index (PI) = (S-D)/M. [S = systole, D = diastole, and M = mean maximum Doppler-Shift frequency over the cardiac cycle]. A repeated measure ANOVA was used to detect differences between mean values of each Doppler index for every week of gestation using the Fisher test (InfoStat V1.5, FCA, Universidad de Córdoba, Córdoba, Argentina). Correlation between Doppler indices was also calculated. All data are shown as mean ± S.D. Results The duration of pregnancy was 148 ± 1.5 days, all lambs were born naturally without any type of assistance. All three Doppler indices were highly correlated: S/D versus RI versus PI, r > 0.84. From weeks 4 to 9 of pregnancy, blood flow was characterized by a systolic pattern (i.e. high resistance with absence of diastolic flow). At 10 and 12 weeks of gestation, 50% and 100% of the fetuses showed a diastolic flow consistent with low resistance, respectively. All three resistance indices decreased (by > 45%, P < 0.05) from week 10 (SD = 7.62 ± 1.82; RI = 0.86 ± 0.03; PI = 1.70 ± 0.23) to week 16 (SD = 2.72 ± 0.52; RI = 0.62 ± 0.07; PI = 0.95 ± 0.18) of pregnancy, with no substantial changes thereafter (P > 0.05). Conclusion Umbilical artery blood flow pattern in ewes was initially systolic (high resistance) but became diastolic (low resistance) from week 11 of pregnancy onwards. Noninvasive Doppler sonography was useful for assessment of umbilical blood flow from 4 to 20 weeks of pregnancy, these reference values may be useful for assessing placental function in high-risk pregnancies e.g. cloned derived pregnancies. P179 Estrogen and Progesterone Receptors in the vagina of progesterone primed and GnRH treated anoestrous ewes Tasende, C*, Acuña, S; López, C; Garófalo, E Cellular and Mollecular Biology, Faculty of Veterinary Medicine, Uruguay The Estrogen and Progesterone Receptors (ER, PR) concentrations were investigated in vagina of anestrous Corriedale ewes treated with GnRH or with Progesterone (P) plus GnRH. Twenty two ewes were assigned to two groups: GnRH (n = 11) and P+GnRH (n = 11). The P+GnRH ewes were treated with 0.33 g of P (CIDR) for 10 Days and immediately after CIDR removal they were treated every 2 h with 6.7 ng of GnRH (i.v.) for 16 h, followed by bolus injection of GnRH (4 μg, Day 0) at 18 h. The GnRH ewes were treated according to the same protocol without P pre-treatment. Ewes were killed on Day 1 (n = 6, for each treatment) and Day 5 (n = 5, for each treatment) after bolus injection. Samples of vagina and blood were taken for receptors and P determinations when the ewes were killed. The ER and PR determinations were performed by ligand binding assay. The ligands used were 3H-E2 for ER or 3H-ORG-2058 for PR, while nonlabelled ligands were diethylstilbestrol and ORG-2058 respectively. The ER, PR and P concentration were analyzed by ANOVA. On Day 5 the P concentration (mean ± pooled s.e.) were higher in the P+GnRH ewes than in the GnRH ewes (6.5±0.54 vs. 3.1±0.61 nmol/L, respectively, P
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Reproduction in Domestic Animals - Facultad de Ciencias Veterinarias

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