Corpus Luteum Blood Flow Research Articles

Conceptus-induced changes in the gene expression of blood immune cells and the ultrasound-accessed luteal function in beef cattle: how early can we detect pregnancy?

We aimed to identify the functional characteristics of the corpus luteum (CL) by color Doppler ultrasonography and changes in interferon-stimulated gene (ISG) expression in peripheral blood mononuclear cells (PBMCs) during early pregnancy in beef cows. We then aimed to use these features to establish earlier pregnancy diagnosis methods. In experiment 1, the CL size and blood flow were accessed by Doppler ultrasonography, and the PBMCs were isolated on Days 8, 12, 15, 18, 20, 22, 25, 30, 45, and 60 post-timed artificial insemination (TAI) from pregnant (n = 10) and nonpregnant cows (n = 12). The abundance of ISG (OAS1, MX1, MX2, and ISG15) transcripts was measured by quantitative PCR. Analyses of OAS1 and MX2 expression in isolated PBMCs (ISG-PBMC method) and Doppler imaging of CL (Doppler-US method) were performed to test the accuracy of these methods for the diagnosis of pregnancy on Day 20 post-TAI (n = 110; experiment 2). In experiment 1, the luteal volume and blood flow were reduced in nonpregnant cows during the first weeks post-TAI, but an evaluation of CL vascularization and size was efficient in identifying nonpregnant cows on Day 20 post-TAI. The expression of ISGs in PBMCs can be stimulated by the presence of a viable conceptus between Days 15 and 22 post-TAI, and the expression of these genes reaches a peak on Day 20. In experiment 2, the Doppler-US and ISG-PBMC methods resulted in similar specificity (85.5 and 87.7%, respectively). However, only the Doppler-US method resulted in 100% sensitivity. In conclusion, the greatest abundance of ISGs in PBMCs and a high detection of luteolysis by Doppler imaging on Day 20 post-TAI can be feasibly used for the earlier detection of nonpregnant cows in reproductive programs. The level of accuracy for our described pregnancy methods is high on Day 20 (80%-91%), but only the Doppler imaging method results in an absence of false-negative diagnoses.

Early pregnancy diagnosis on days 18 to 21 postinsemination using high-resolution imaging in lactating dairy cows

The aim was to assess the ability of corpus luteum (CL) and uterine ultrasound characteristics on d 18 to 21 to predict pregnancy status in lactating dairy cows. Ultrasound examinations were carried out on cows (n=164) on d 18 to 21 following artificial insemination (AI). Images of the uterus and CL were captured using a Voluson i ultrasound device (General Electric Healthcare Systems, Vienna, Austria) equipped with a 12-MHz, multi frequency, linear array probe. Serum concentrations of progesterone were determined from blood samples collected at each ultrasound examination. Images of the CL were captured and stored for calculation of CL tissue area and echotexture. Images of the CL and associated blood flow area were captured and stored for analysis of luteal blood flow ratio. Longitudinal B-mode images of the uterine horns were stored for analysis of echotexture. Diagnosis of pregnancy was made at each ultrasound examination based on CL blood flow, CL size, and uterine echotexture. Pregnancy was confirmed by ultrasonography on d 30 after AI. The relationship between ultrasound measures and pregnancy outcome, as well as the accuracy of the pregnancy diagnosis made at each ultrasound examination was assessed. Progesterone concentrations and CL tissue area were greater in pregnant compared with nonpregnant cows on all days. The CL blood flow ratio was higher in pregnant compared with nonpregnant cows on d 20 and 21 after AI. Echotexture measures of the CL and uterus were not different between pregnant and nonpregnant cows on any day of examination. The best logistic regression model to predict pregnancy included scores for CL blood flow, CL size, and uterine echotexture on d 21 following AI. Accuracy of pregnancy diagnosis was highest on d 21, with sensitivity and specificity being 97.6 and 97.5%, respectively. Uterine echotexture scores were similar for pregnant and nonpregnant cows from d 18 to 20. On d 21, pregnant cows had higher uterine echotexture scores compared with nonpregnant cows. The logistic regression equation most likely to provide a correct pregnancy diagnosis in lactating dairy cows included the visual score for CL blood flow, CL size, and uterine echotexture on d 21 after AI. In support of this finding, the diagnostic accuracy for visual scores of CL blood flow, CL size, and uterine echotexture were also highest on d 21.

Corpus Luteum Development and Function after Supplementation of Long‐Acting Progesterone During the Early Luteal Phase in Beef Cattle

Strategic supplementation of P4 may be used to increase conception rates in cattle, but timing of supplementation in relation to ovulation, mass of supplementary P4 and formulation of the P4-containing supplement has not been determined for beef cattle. Effects of supplementation of long-acting progesterone (P4) on Days 2 or 3 post-ovulation on development, function and regression of corpus luteum (CL) were studied in beef cattle. Cows were synchronized with an oestradiol/P4-based protocol and treated with 150 or 300mg of long-acting P4 on Day 2 or 3 post-ovulation (6-7 cows/group). Colour-doppler ultrasound scanning and blood sample collection were performed from Day 2-21.5. Plasma P4 concentrations were greater (p<0.05) from Day 2.5-5.5 in the Day 2-treated groups and from Day 3.5-5.5 in the Day 3-treated cows than in the control group. CL area and blood flow during Day 2-8.5 did not differ (p>0.05) among groups, suggesting no effect of P4 treatment on luteal development. The frequency of cows that began luteolysis before Day 15 was greater (p<0.04) in cows treated with 300mg than in the controls, but there were no differences between non-treated and 150mg-treated cows. The interval from pre-treatment ovulation to functional and structural luteolysis was shorter (p<0.01) in the combined P4-treated groups than in the control cows. In conclusion, was showed for the first time that long-acting P4 supplementation on Day 2 or 3 post-ovulation increases P4 concentrations for ≥3day, has no effect on luteal development, but anticipates the beginning of luteolysis in beef cattle.

Color Doppler flow imaging for the early detection of nonpregnant cattle at 20 days after timed artificial insemination

The objective was to determine the accuracy of a pregnancy test for predicting nonpregnant cattle based on the evaluation of corpus luteum (CL) blood flow at 20d (CLBF-d20) after timed artificial insemination (TAI). Crossbred Holstein-Gir dairy heifers (n=209) and lactating cows (n=317) were synchronized for TAI using the following protocol: intravaginal implant (1.0g of progesterone) and 2mg of estradiol benzoate i.m. on d −10, implant removal and 0.526mg of sodium cloprostenol i.m. on d −2, 1mg of estradiol benzoate i.m. on d −1, and TAI on d 0. On d 20, animals underwent grayscale ultrasonography (US) to locate the CL and color flow Doppler to evaluate CLBF-d20 using a portable ultrasound equipped with a 7.5-MHz rectal transducer. Based only on a visual, subjective CLBF evaluation, the animals were classified as pregnant or not pregnant. On d 30 to 35, blinded from results of the previous diagnosis, the same operator performed a final pregnancy diagnosis using US to visualize the fetal heartbeat (gold standard; US-d30). A second evaluator also analyzed the CLBF-d20 in the same animals by watching 7-s recorded videos. Blood samples were collected from a subset of 171 females to determine, by RIA, plasma progesterone (P4) concentrations, which indicate CL function. The final pregnancy outcome (US-d30) was retrospectively compared with the CLBF-d20 diagnoses and then classified either as correct or incorrect. The number of true positive, true negative, false positive, and false negative decisions were inserted into a 2×2 decision matrix. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the CLBF-d20 test were calculated using specific equations. Binomial variables (pregnancy rate and proportions) were analyzed using Fisher’s exact test for the effect of parity and to compare between evaluators and tests (CLBF-d20 vs. plasma P4). The kappa values were calculated to quantify the agreement between CLBF-d20 and the gold standard (US-d30) and between evaluators. The performance parameters of CLBF-d20 test were as follows: sensitivity=99.0%, specificity=53.7%, positive predictive value=65.1%, negative predictive value=98.5%, and accuracy=74.8%. False negatives represented only 0.4% of the exams. No differences existed in these parameters between evaluators (no. 1 vs. no. 2) and tests (CLBF-d20 vs. plasma P4). Moreover, a high level of agreement was observed between evaluators (0.91). In conclusion, visual evaluation of CLBF-d20 represents a quick, reliable, and consistent diagnostic test that enables the early detection of nonpregnant cattle.

Effect of dinoprost and cloprostenol on serum nitric oxide and corpus luteum blood flow during luteolysis in ewes

In this study we compared the effect of dinoprost and cloprostenol on changes of corpus luteum blood flow during luteolysis. Ten nonlactating cyclic ewes were synchronized with double PGF2α injections 11 days apart. At Day 10, the animals were classified into 2 groups and received the third dose of PGF2α after confirmation of the presence of a mature CL. The first group received (12.5 mg/im) dinoprost and the second group received (250 μg/im) cloprostenol. A color Doppler ultrasound scan was performed by the same operator according to the following timeline: 0, 0.5, 1, 2, 4, 6, 12, and 24 hours, then every 24 hours until Day 4). The size, morphology, and blood flow of the CL was evaluated during the regression. The results showed that regression of the CL did not differ between the dinoprost and cloprostenol groups. There was no significant effect on diameter of the CL in both groups, though the size of the CL decreased gradually and slowly. Pretreatment progesterone concentration did not differ between groups. The results showed that the nitric oxide level was significantly increased within half an hour after the dinoprost treatment, and was significantly decreased in the cloprostenol group after half an hour. The blood velocity was increased significantly half an hour after the dinoprost treatment and it was decreased in the cloprostenol-treated group. In conclusion, both cloprostenol and dinoprost affect CL by controlling the nitric oxide level and blood supply of the CL via different mechanisms to induce luteolysis.

173 USE OF CORPUS LUTEUM AREA AS A PREDICTOR OF ONGOING FUNCTIONAL LUTEOLYSIS IN DAIRY HEIFERS

The beginning of functional luteolysis in cattle ranges from Days 16–19 (Day 0 = ovulation). During functional luteolysis, luteal size and blood flow decrease in heifers. The wide range of beginning of luteolysis and the low sensitivity of quick progesterone (P4) assays preclude targeting an individual for a specific period of luteolysis. The present study aimed to evaluate the changes in area and blood flow of corpus luteum (CL) during spontaneous luteolysis and test the hypothesis that a decrease in CL area (cm2) is more efficient in targeting the luteolytic period than using a specific day postovulation. Blood sampling and ultrasound scanning of CL were done every 12 h from Days 13–22 in dairy heifers (Holstein) and was used for a retrospective study (Experiment 1; n = 6 heifers). The P4 concentrations, maximum CL area, and the percentage of CL area with coloured signals of blood flow were centralized to 12-h sample when the end of luteolysis (P4 &lt; 1 ng mL–1) was detected. The percentage decrease in CL area and blood flow between Day 13 and the 12-h sample before the end of luteolysis were calculated. In Experiment 2, a blood sampling and transrectal ultrasonography of CL were performed every 8 h starting on Day 14 to determine the beginning and end of functional luteolysis (n = 20 heifers). The hour of detection of luteolysis was defined as the scan at 8-h intervals when maximum CL area decreased by 25 or 12.5% on Day 14. Based on P4 concentrations, the preluteolytic, luteolytic, and postluteolytic periods in each heifer were identified. The area and blood flow were measured by a duplex B-mode and pulse-wave colour Doppler ultrasound instrument. In Experiment 1, the retrospective study indicated a 25% decrease in CL area and 65% decrease in blood flow 12 h before the end of functional luteolysis. In Experiment 2, the 25% and 12.5% CL area decrease from Day 14 occurred, respectively, on Days 18.1 ± 0.6 and 17.2 ± 0.5 postovulation. The hypothesis that a decrease in CL area is more efficient in targeting ongoing luteolysis than using a specific day postovulation was supported. This was indicated by the greater (P &lt; 0.05) frequency of heifers detected in luteolytic period (12 of 20 heifers) using the criteria of a 12.5% decrease in CL area than using a specific day (Table 1). Using a day postovulation for obtaining heifers in the luteolytic phase was only 0 to 30% effective for targeting luteolysis. In conclusion, the technique of using an ultrasonic 12.5% reduction in CL area is a good tool to detect ongoing luteolysis and may be useful as an experimental design to study factors related to the luteolytic period in cattle. Table 1.Percentage values of heifers in preluteolytic, luteolytic, or postluteolytic periods according to a decrease in CL area criteria or a day postovulation Supported by grants from FAPESP (2012/04004-8 to GP).

P11.04: Correlation between ultrasound corpus luteum blood flow and serum progesterone concentrations of infertile women in Medan, Indonesia

P11.04: Correlation between ultrasound corpus luteum blood flow and serum progesterone concentrations of infertile women in Medan, Indonesia

Quantification of blood volume and vascular perfusion in the corpus luteum of rhesus macaques during the menstrual cycle

OBJECTIVE: To determine changes to luteal vascular flow and volume in rhesus monkeys using dynamic contrast enhanced ultrasound (d-CEU). DESIGN: In vivo, repeated measures. MATERIALS AND METHODS: Hormonal status of 8 adult cycling female macaques was monitored to detect the day of ovulation. D-CEU with Definity (Lantheus) microbubble perfusion was performed on the corpus luteum (CL) at the early, mid, mid-late and late luteal phase (LP; Mol Reprod Dev, 76: 367-378, 2009), as recently described for the uterus (Fertil Steril 95:1316, 2011). Vascular parameters were quantified in d-CEU video captures using a region of interest (ROI) focused on the CL. Video intensity (VI) was normalized to brachial artery flow to provide an estimate of blood volume (BV); change in VI (DVI/sec) was used to determine vascular perfusion rate (VP). RESULTS: Significant changes in BVand VP (P<001 and P1⁄40.07) were observed between stages of the LP. BV was negatively correlated with luteal day (P<0.02). In contrast, serum progesterone (P) levels correlated positively with BV (P1⁄40.04) and VP (P1⁄40.001). P area under curve (AUC) also positively correlated with AUC for BV (P1⁄40.03). In typical LPs, BV was highest during the early LP, before decreasing 3-fold in late LP CL. In contrast, VP peaked at mid LP, with values 5 and 11-fold greater than early and late LP, respectively. Two females were identified with luteal insufficiency based on peak P levels during the mid and mid-late LP (2.3 0.2 ng/ml), whereas 5 had normal LPs (6.2 1.2 ng/ml). Another female was identified with a luteal cyst on the contralateral ovary, and a functional CL that regressed before P levels declined. Insufficient LPs trended toward lower peak BVand VP, compared to typical LPs. CONCLUSION: D-CEU provides a non-invasive measurement of the dynamics of CL blood volume and flow during the luteal lifespan in the menstrual cycle. Reduced vascular perfusion is evident in animals with reduced P secretion and in animals with aberrant luteal function. Supported by: P51OD011092, R01HD020869 (RLS).

Corpus luteum development and function and relationship to pregnancy during the breeding season in the Mediterranean buffalo

The aim of this study was to ascertain corpus luteum (CL) development and function in buffaloes synchronized and mated by artificial insemination (AI) during the breeding season. Italian Mediterranean buffalo cows (n = 43) at 86.5 ± 2.7 days postpartum were synchronized by the Ovsynch-TAI Program and inseminated using frozen thawed semen at 20 and 44 h after the second injection of GnRH. The CL dimensions (diameter and area) and blood flow were examined on Days 5, 10, 15, 20, and 25 after AI by realtime B-mode/colour-Doppler ultrasonography. The resistive index (RI), pulsatility index (PI) and time average medium velocity (TAMV) were recorded at each time, together with CL dimensions. Blood samples were taken on the days of ultrasonography for progesterone (P4) assay by RIA. Data were grouped into pregnant or non-pregnant and retrospectively analyzed by repeated measure ANOVA and correlation analyses. Dimensions of the CL on Days 10, 20, and 25 after AI were greater (P < 0.01) in buffaloes pregnant on Day 45 (n = 18) compared with non-pregnant buffaloes (n = 25). The former buffaloes also showed a greater (P < 0.01) rate of CL growth between Days 5 and 10 after AI. Blood flow to the CL on Day 10 after AI showed a higher TAMV (P < 0.01) and lower RI (P < 0.05) in pregnant buffaloes compared with non-pregnant buffaloes. Negative correlations were observed on Day 10 after AI between CL diameter and RI (r = −0.61; P < 0.01) and PI (r = −0.60; P < 0.01); P4 concentrations and RI (r = −0.46; P < 0.02); and RI and pregnancy (r = 0.45; P < 0.02). Positive correlations were observed between pregnancy and CL size (r = 0.54; P < 0.01), ΔCL diameter between Days 5 and 10 (r = 0.52; P < 0.01), ΔCL area between Days 5 and 10 (r = 0.48; P < 0.015), and ΔP4 between Days 5 and 10 (r = 0.50; P < 0.01). Based on these findings it is concluded that the period between Day 5 and 10 is very important for CL growth and crucial in evaluating pregnancy. Accordingly, the assessment of CL parameters during the period from Day 5 to Day 10 after AI might be used to predict the likelihood of an ongoing pregnancy.

Effect of oxytocin infusion on luteal blood flow and progesterone secretion in dairy cattle

The objective of this study was to investigate the effects of oxytocin infusion on corpus luteum (CL) function during early to mid-diestrus by measuring luteal size (LS) and luteal blood flow (LBF) along with plasma levels of progesterone (P4) and prostaglandin metabolites (13,14-dihydro-15-keto-prostaglandin F2α, PGFM). On day (D) 7 of the estrus cycle (D1 = ovulation), seven cows received 100 IU of oxytocin (OXY) or placebo (PL) following a Latin square design. LS and LBF increased in both groups over time and no differences were observed between the groups. PGFM did not differ either within the groups over time or between the groups at any time point. P4 of the OXY group was higher compared to that of the the PL group 360 min after the infusion (p = 0.01) and tended to be higher at the time points 450 min, 48 h, and 72 h (all p = 0.08). Results from this study support the hypothesis that OXY is not directly involved in the mechanism(s) governing blood flow of the CL and has no remarkable effects either on luteal size or P4 and PGFM plasma levels. Further investigation is needed to elucidate the role of OXY in CL blood flow during early and late luteal phases.

Efficacy of PGF2α on Pre‐ovulatory Follicle and Corpus Luteum Blood Flow

The aim of this study was to evaluate the effect of cloprostenol administration on the blood flow of pre-ovulatory follicle (PF) and corpus luteum (CL), progesterone secretion and pregnancy outcome in buffaloes subjected to AI. The trial was performed on 75 Italian buffaloes at 182 ± 8 days in milk. Synchronized animals were randomly divided into two groups on the day of oestrus: Group T (n = 37) received a 0.524 mg intramuscular injection of cloprostenol and Group C (n = 38) received saline. Ultrasound examinations of the ovaries were performed 5 h after AI on the PF and 10 and 20 days after AI on the CL. Resistive (RI) and pulsatily index (PI) were calculated by colour-Doppler mode in each examination. Blood samples were collected on days 10, 20 and 25 after AI for progesterone assay and 25 days after AI, ultrasonography was performed to assess pregnancy, which was confirmed on day 45. Subjects pregnant on day 25 but not on day 45 were considered to have undergone late embryonic mortality (LEM). Statistical analysis was performed by anova. No differences were found in PF dimensions, CL size and blood flow on day 10 and 20 after AI between treated and control groups. Pre-ovulatory follicle area was higher in buffaloes that resulted pregnant on day 25 after AI compared to those that were non-pregnant (2.13 vs 1.66 cm in pregnant and non-pregnant buffaloes, respectively), while non-pregnant buffaloes showed higher values of RI (0.49 vs 0.30; p < 0.05) and PI (1.0 vs 0.37; p = 0.07) compared to pregnant subjects. Treatment by cloprostenol did not influence pregnancy rate both on day 25 (31/75; 41.3%) and 45 (27/75; 36.0%), progesterone levels and incidence of LEM (4/31; 12.9%). In conclusion, cloprostenol administration at the time of AI does not seem to affect PF and CL blood flow.

Corpus luteum Function and Pregnancy Outcome in Buffaloes during the Transition Period from Breeding to Non‐Breeding Season

The aim in this study was to investigate corpus luteum function and embryonic loss in buffaloes mated by artificial inseminations (AI) during the transitional period from breeding to non-breeding season. The study was carried out using 288 multiparous Italian Mediterranean Buffalo cows at 110 ± 4 days in milk. The buffaloes were mated by AI after synchronization of ovulation by the Ovsynch-TAI protocol 25 days after AI buffaloes underwent trans-rectal ultrasonography to assess embryonic development. Pregnancy diagnosis was confirmed on Days 45 and 70 after AI by rectal palpation. Buffaloes pregnant on Day 25 but not on Day 45 were considered to have undergone late embryonic mortality (LEM), whilst buffaloes pregnant on Day 45 but not on Day 70 were considered to have undergone foetal mortality (FM). Corpus luteum size and blood flow were determined by real-time B-mode/colour-Doppler on day 10 after AI in 122 buffaloes. The resistive index (RI) and pulsatility index (PI) were recorded at the time. Milk samples were collected on Days 10, 20 and 25 after AI in all inseminated buffaloes for the assay of whey P4 concentrations. Data were analysed by anova. Pregnancy rate on Day 25 after AI was 48.6% (140/288) and declined to 35.4% (102/288) and 30.6% (88/288) by Day 45 and Day 70 respectively. The incidences of LEM and FM were respectively 27.1% (38/140) and 13.7% (14/102). Pregnant buffaloes had greater (p < 0.01) whey concentrations of P4 from Day 20 onwards than buffaloes which showed LEM, whilst P4 in buffaloes that showed FM did not differ from the other two groups on Day 10 and Day 20. Corpus luteum blood flow on Day 10 after AI showed higher RI (p < 0.05) and PI (p = 0.07) values in buffaloes that subsequently were not pregnant on Day 25 compared with pregnant buffaloes. Buffaloes that were not pregnant on Day 45 also had a higher (p = 0.02) RI value on Day 10 than pregnant buffaloes, whilst PI values on Day 10 did not differ for the two groups of buffaloes. It was concluded that blood flow to the corpus luteum on Day 10 after AI influences corpus luteum function as judged by P4 secretion and also embryonic development and attachment in buffaloes.

Luteal blood flow and luteal function.

BackgroundBlood flow in the corpus luteum (CL) is associated with luteal function. The present study was undertaken to investigate whether luteal function can be improved by increasing CL blood flow in women with luteal phase defect (LFD).MethodsBlood flow impedance in the CL was measured by transvaginal color-pulsed-Doppler-ultrasonography and was expressed as a resistance index (RI). The patients with both LFD [serum progesterone (P) concentrations < 10 ng/ml during mid-luteal phase] and high CL-RI (≥ 0.51) were given vitamin-E (600 mg/day, n = 18), L-arginine (6 g/day, n = 14) as a potential nitric oxide donor, melatonin (3 mg/day, n = 13) as an antioxidant, or HCG (2,000 IU/day, n = 10) during the subsequent menstrual cycle.ResultsIn the control group (n = 11), who received no medication to increase CL blood flow, only one patient (9%) improved in CL-RI and 2 patients (18%) improved in serum P. Vitamin-E improved CL-RI in 15 patients (83%) and improved serum P in 12 patients (67%). L-arginine improved CL-RI in all the patients (100%) and improved serum P in 10 patients (71%). HCG improved CL-RI in all the patients (100%) and improved serum P in 9 patients (90%). Melatonin had no significant effect.ConclusionVitamin-E or L-arginine treatment improved luteal function by decreasing CL blood flow impedance. CL blood flow is a critical factor for luteal function.

The evaluation of corpus luteum blood flow using color-flow Doppler ultrasound for early pregnancy diagnosis in bovine embryo recipients

The objective of this experiment was to evaluate corpus luteum blood flow (CLBF) as an early indicator of pregnancy status in bovine embryo recipients. Fifty crossbred beef cows were submitted to embryo transfer on Day 7 after estrus. On Days 7, 11, 13, 15, 17, 19, 21, 26, 33, and 40, a blood sample was taken, the CL examined using a color-flow Doppler ultrasound scanner, and video was recorded of each scanning session. Ultrasound data were grouped by the first day progesterone concentrations were <1 ng/mL (indicating early embryo loss, EEL) through Day 21 (EEL-17, n = 3; EEL-19, n = 9; EEL-21, n = 3), absence of an embryo on Days 26, 33, or 40 (late embryo loss; LEL; n = 12), or remained pregnant ( P; n = 23). The first decrease in CLBF of EEL-17, EEL-19, and EEL-21 cows compared to P cows occurred on Days 17, 19, and 21, respectively ( P < 0.05). There was no difference in CLBF between LEL and P cows on Days 17, 19, and 21. Six evaluators diagnosed pregnancy from randomized video clips on Days 17, 19, and 21. Evaluators made more ( P < 0.004) correct diagnoses on Day 19 than Day 17. Sensitivity (82.9 ± 10.1%) was not affected by day. From Days 17 to 19, diagnostic specificity increased ( P = 0.046) from 43.2 ± 3.0 to 54.3 ± 3.0% but remained unchanged thereafter. Due to low specificity and sensitivity, evaluation of CLBF alone was insufficient for early pregnancy diagnosis.

Reduction in Circulating Estradiol by Ablation of Follicles Delays Luteolysis in Heifers.

Circulating estradiol (E2) and progesterone (P4) may regulate timing of prostaglandin F2α (PGF2α) release from the uterus. We hypothesized that removal of circulating E2 by ablating follicles >4 mm would therefore delay luteolysis. On Day 9 (Day 0 = ovulation), Holstein heifers were randomized by replicate into three groups. Follicle ablations (all follicles ≥ 4 mm ablated) were done either once on Day 9 (FA9; n=6), on Days 9, 11, 13 and 15 (FA9-15; n=6), or on Days 9, 11, 13, 15, 17, 19 and 21 (FA9-21; n=7). A duplex B-mode and pulsed-wave color-Doppler ultrasound scanner equipped with a linear array 7.5 MHz transrectal transducer was used to estimate corpus luteum (CL) blood flow, based on percentage of the CL with color-Doppler signals during real time scanning. Blood samples were collected daily and plasma assayed by RIA for P4 and E2 concentrations. The CL blood-flow and hormone data were analyzed by SAS MIXED procedure with a REPEATED statement for main effects of group and day and their interaction. Both main effects and the interaction were significant (P<0.01) for P4, E2, and CL blood flow. Mean P4 concentrations first began to decrease (P<0.02) on Day 14 in FA9 and were lower (P<0.05) than in the other two groups on Days 15 and 16. In FA9-15, P4 first began to decline on Day 16 (P<0.01) and was lower (P<0.05) than in FA9-21 on Days 17, 18 and 19. In FA9-21, the first P4 decrease (P<0.01) occurred on Day 19, and there was no significant difference among groups after Day 19. For CL blood flow, the first decrease (P<0.05) began in FA9 on Day 15 and the values were lower (P<0.05) than in FA9-15 on Day 16. In FA9-15, the first decrease (P<0.05) in blood flow occurred on Day 17 and the values were lower (P<0.05) from Day 17 to Day 21 than in FA9-21. The first decrease (P<0.05) in CL blood flow for FA9-21 occurred on Day 19. In FA9, the E2 concentration began to increase (P<0.01) on Day 13 and the values were higher (P<0.05) from Day 14 to Day 17 than in FA9-15 and FA9-21. In FA9-15, the first increase (P<0.01) in E2 occurred on Day 16, whereas in FA9-21 the first increase (P<0.04) occurred on Day 22. Follicular ablation delayed the rise in circulating E2 with earlier (P<0.002) mean peak E2 concentrations in FA9 (17.6±0.7 days after ovulation) than in FA9-15 (20.3±0.3 d) and FA9-21 (24.9±0.3 d). In FA9 and FA9-15 luteolysis did not begin until after the last ablation and 1 day after the first continuous increase in E2 plasma concentration, but in FA9-21 luteolysis began 3 days before the last ablation and first continuous increase in E2 concentration. The first significant decreases in blood flow occurred one day after the first significant decrease in P4 (FA9) or both occurred on the same day (FA9-15, FA9-21). Sequential ablations of all follicles (≥4 mm) delayed both functional (decreased P4) and structural (decreased CL blood flow) indicators of luteolysis. Thus, normal follicle growth was essential for normal timing of luteolysis in heifers. It seems likely that the delay in luteolysis caused by follicular ablation was due to reduced circulating E2, but luteolysis occurred before a detectable increase in plasma E2 concentration when the CL was maintained for long period as in the FA9-21 group.

Temporal Associations among Pulses of 13,14-Dihydro-15-keto-PGF2alpha, Luteal Blood Flow, and Luteolysis in Cattle1

Luteal blood flow was studied in heifers by transrectal color-Doppler ultrasound. Data were normalized to the decrease in plasma progesterone to <1 ng/ml (Day 0 or Hour 0). Blood flow in the corpus luteum (CL) was estimated by the percentage of CL area with color flow signals. Systemic prostaglandin F2alpha (PGF) treatment (25 mg; n=4) resulted in a transient increase in CL blood flow during the initial portion of the induced decrease in progesterone. Intrauterine treatment (1 or 2 mg) was done to preclude hypothetical secondary effects of systemic treatment. Heifers were grouped into responders (luteolysis; n=3) and nonresponders (n=5). Blood flow increased transiently in both groups; induction of increased blood flow did not assure the occurrence of luteolysis. A transient increase in CL blood flow was not detected in association with spontaneous luteolysis when examinations were done every 12 h (n=6) or 24 h (n=10). The role of PGF pulses was studied by examinations every hour during a 12-h window each day during expected spontaneous luteolysis. At least one pulse of 13,14-dihydro-15-keto-PGF2alpha (PGFM) was identified in each of six heifers during the luteolytic period (Hours -48 to -1). Blood flow increased (P<0.02) during the 3-h ascending portion of the PGFM pulse, remained elevated for 2 h after the PGFM peak, and then decreased (P<0.03) to baseline. Results supported the hypothesis that CL blood flow increased and decreased with individual PGFM pulses during spontaneous luteolysis.

Effect of Intraluteal Injection of Endothelin Type A Receptor Antagonist on PGF2.ALPHA.-induced Luteolysis in the Cow

Endothelin-1 (ET-1) is a luteolytic mediator in the bovine corpus luteum (CL), and its action appears to be via endothelin type A receptor (ETR-A). Thus, the aim of the present study was to determine the effect of ETR-A antagonist on PGF2alpha-induced luteolysis in the cow. Cows on days 10-12 of the estrous cycle were subjected to five intraluteal injections of the ETR-A antagonist LU 135252 in saline or only saline at -0.5, 2, 4, 6, and 8 h after PGF2alpha administration (=0 h). Serial luteal biopsies were conducted to determine the expression of mRNA in the luteal tissue. There were no significant differences in the decrease in plasma progesterone (P) concentrations and the mRNA expressions of steroidogenic acute regulatory protein and 3beta-hydroxysteroid dehydrogenase/Delta5, Delta4-isomerase between the ETR-A antagonist-treated group and the control group. However, the start of the decline in CL volume and blood flow area surrounding the CL was delayed for almost two days in the ETR-A antagonist-treated group compared to the control group. The mRNA expression of preproET-1 and endothelin type B receptor increased in both groups, while the ETR-A mRNA remained unchanged. In addition, caspase-3 mRNA expression increased significantly at 24 h in the control group only and its level was higher than that of the ETR-A antagonist-treated group. Thus, the present study suggests that ET-1 regulates structural luteolysis via ETR-A by controlling blood vessel contraction in the CL of the cow.

Power and colour Doppler ultrasonography for the evaluation of the vasculature of the human corpus luteum.

We used power Doppler imaging to examine neovascularization in the corpus luteum (CL) in 12 healthy volunteers. We also investigated whether CL blood flow reflected luteal function. The ratio of the area of vessels in the CL to the area of a sectional plane at the maximum diameter of the CL observed by power Doppler (FA ratio) was used as a quantitative index of the vascularity of the CL. The pulsatility index (PI) was significantly lower in ovarian arteries with CL than without CL (P < 0.05). Changes in ovarian arterial and intra-luteal PI appeared to reflect physiological changes in the vasculature of the CL. There was no correlation between the volume of the CL or the FA ratio and the concentration of progesterone. The pattern of changes in the product of the FA ratio and the CL volume and in the progesterone concentration was similar. The progesterone concentration was positively correlated with this product (r = 0.74, P < 0.01). The product of the FA ratio and the CL volume plateaued during the mid- to late luteal phase, suggesting the presence of functional and structural luteolysis. These findings suggest that colour Doppler ultrasonography, including power Doppler imaging, can detect physiological changes in the blood flow of the ovary in the luteal phase, and may be a useful noninvasive tool for evaluating CL function.

The assessment of normal and abnormal luteal function by transvaginal color Doppler sonography

Objective: To evaluate intraovarian resistance index (RI) in 47 healthy fertile volunteers with ovulatory cycles, 28 patients with luteal phase defect (LPD) and four patients with luteinized unruptured follicle (LUF Sy). Study design: Transvaginal color Doppler assessment of the follicular and corpus luteum blood flow and plasma progesterone (P) levels were obtained in each patient. Results: Significantly higher intraovarian artery RI (< 0.001) was obtained for LPD group than for controls during the luteal phase. In the control group both follicular and corpus luteum RI were significantly lower ( P < 0.001) on the dominant side, while in LPD group no difference ( P > 0.05) between the sides occurred. Mean P levels were significantly lower ( P < 0.001) in the LPD group (6.9 ± 2.3 ng/ml) than in controls (24.1 ± 11.4 ng/ml). In all the LPD patients histopathology revealed delayed endometrial pattern, while normal endometrial dating was found in all the evaluated patients from the control group ( n = 15). In the patients with LUF Sy ( n = 4) similar RI values were obtained in the follicular and corpus luteum phase. There was no difference between the sides in terms of the intraovarian RI. while subnormal values of P were obtained in all the examined patients (14.1 ± 6.2 ng/ml). Conclusion: Transvaginal color Doppler may predict the functional capacity of the corpus luteum.

Corpus luteum blood flow in abnormal early pregnancy.

Thirty-eight consecutive patients with the diagnosis of abnormal early singleton pregnancy were studied with transvaginal color velocity imaging and pulsed Doppler ultrasonography to assess corpus luteum blood flow. Mean gestational age was 8.5 weeks (range, 6 to 12 weeks). Patients' mean age was 30.9 years (range, 24 to 43 years). There were 19 (50%) threatened abortions, 13 (34.2%) missed abortions, and six (15.8%) anembryonic pregnancies. Blood flow impedance in the corpus luteum was estimated by calculating the resistive index. The results of this study group were compared with those obtained in a series of 85 normal singleton early pregnancies used as controls. Overall, detection of corpus luteum blood flow was 78.9% in study group and 76.4% in control group (P = 0.51). Mean resistive index +/- standard deviation in the control group was 0.50 +/- 0.10; the corresponding values in threatened abortion, anembryonic pregnancy, and missed abortion were 0.52 +/- 0.10, 0.42 +/- 0.06, and 0.57 +/- 0.05, respectively. No statistical. differences in mean resistive index were found among threatened abortion and anembryonic pregnancy with respect to control group. A higher mean resistive index was observed in missed abortion than in control group (P < 0.01). In conclusion, our data show that luteal vascularization might be decreased in missed abortion but not in threatened abortion and anembryonic pregnancy.