Detection of early placental hormone production in embryo transfer cycles lacking a corpus luteum

The present study was conducted to define the action of a mixture obtained by the extraction and purification of real fly ash, on specific toxicity endpoints, such as hormonal secretion, CYP1A1 expression, DNA damage and cell apoptosis. JEG-3 cell line was exposed in vitro to different doses of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) or Polychlorinated dibenzo-p-dioxin/Polychlorinated dibenzo-P-furan (PCDD/PCDF) mixture. Both TCDD and the mixture decreased hCG secretion, while inhibition of progesterone levels was noted only under the influence of TCDD. The changes in hormone production were not due to the action on cell viability. There were time-dependent differences in CYP1A1 expression in cells exposed to TCDD and PCDD/PCDF mixture. Both TCDD and PCDD/PCDF mixture did not induce the DNA damage, as evaluated by the comet assay. Significantly lower DNA migration from the head of comet into the comet tail was noted after the removal of reagents. The highest efficiency of this process was noted 4 h after the TCDD and 24 h after the PCDD/PCDF mixture removal. These results suggest that the DNA adducts and/or DNA—DNA cross-links were formed. Neither TCDD nor PCDD/PCDF mixture had any effect on cell apoptosis assessed by caspase-3 activity and Hoechst 33258. Taken together, these findings clearly indicate a weaker action of the mixture when compared with TCDD. However, in both cases, their action was not due to the induction of the DNA damage and subsequent cell apoptosis but due to a direct influence of these toxicants on placental hormone production. Human & Experimental Toxicology ( 2007) 26, 407—417

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108 FACTORS AFFECTING PREGNANCY RATES IN RECIPIENTS RECEIVING IN VITRO-PRODUCED EMBRYOS BY FIXED TIME EMBRYO TRANSFER

Reproduction Fertility and Development ◽

10.1071/rdv28n2ab108 ◽

2016 ◽

Vol 28 (2) ◽

pp. 184

Author(s):

M. Pelizzari ◽

A. Tribulo ◽

J. Garzon ◽

B. Bernal ◽

R. Tribulo ◽

...

Keyword(s):

Corpus Luteum ◽

Embryo Transfer ◽

Body Condition Score ◽

Prostaglandin F2α ◽

Fixed Time ◽

Pregnancy Rates ◽

Uterine Horn ◽

Device Removal ◽

Embryo Transfer Technique

A retrospective analysis of factors that affect pregnancy rates from 4214 fresh in vitro-produced (IVP) embryos that were transferred at a fixed-time (FTET) in 20 different farms. Recipients were all cycling cows or heifers that were synchronized with 1 of 3 treatments: 1) treatments with progesterone (P4) devices and 2 mg of oestradiol benzoate (EB) on Day 0 (day of insertion) and 24 h after device removal (Day 8); 2) treatments with P4 devices and EB on Day 0, but with 0.5 mg of oestradiol cypionate (ECP) at device removal (Day 8); or 3) treatments with P4 devices and GnRH on Day 0 and a second GnRH 60 h after device removal (Day 5). Cows in all treatment groups also received 500 µg of cloprostenol (prostaglandin F2α) at the time of P4 device removal and 400 IU of eCG either at device removal or 3 days before device removal. All embryos were transferred 7 or 8 days after the expected time of oestrus (24 h after EB, 48 h after ECP or at the time of the second GNRH for each synchronization treatment, respectively). On the day of embryo transfer, recipients were examined by ultrasonography and those with corpus luteum >14 mm in diameter received a fresh, IVP embryo in the uterine horn ipsilateral to the corpus luteum. Pregnancy rates were determined by ultrasonography 35 days after FTET. Data were analysed by logistic regression. Independent variables were classified into the following three categories. 1) Factors related to the recipient and the environment; there were no significant differences in pregnancy rates for corpus luteum diameter (≥14 and <16 mm, ≥16 and <18 mm, or ≥18 mm; P = 0.46), number of corpus luteum (1 or ≥2; P = 0.26), and category of recipient (cow or heifer; P = 0.21). However, there were significant effects of farm (P = 0.01) and body condition score (BCS; P = 0.01). Cows with BCS ≥4.5 (1 to 5 scale) resulted in lower pregnancy rates (4/20, 20.0%) than those with BCS 2 (74/225, 32.9%), 2.5 (502/1434, 35.0%), 3 (570/1467, 38.9%), 3.5 (193/532, 36.3%), and 4 (44/118, 37.3%). 2) Factors related to the synchronization treatment; there were no significant differences between recipients receiving eCG at device removal (84/209, 40.2%) or 3 days before device removal (874/2291, 38.1%; P = 0.35). However, recipients synchronized with P4 devices and ECP had higher (P = 0.01) pregnancy rates (232/483, 48.0%) than those treated with EB (679/1888, 36.0%) or gonadotropin-releasing hormone (47/129, 36.4%). 3) Factors related to the embryo transfer technique; day of the recipient’s oestrous cycle (P = 0.36), stage of embryo transferred (IETS stages 6 or 7; P = 0.62), and operator (P = 0.57) did not affect pregnancy rates. However transfers made in the anterior third of the uterine horn resulted in higher (649/1545, 42.0%) pregnancy rates than those in the mid-third (845/2511, 33.6%) or in the distal third (6/35, 17.1%; P = 0.01). It was concluded that factors related to the recipient and the environment (farm and BCS), the synchronization treatment (ECP), and the embryo transfer technique (site of deposition) affect pregnancy rates in recipients of embryos produced in vitro and transferred at a fixed time.

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109 RELATIONSHIP BETWEEN ULTRASONIC MORPHOLOGY OF CORPUS LUTEUM IN HOLSTEIN HEIFERS AND PREGNANCY RATE AFTER EMBRYO TRANSFER

Reproduction Fertility and Development ◽

10.1071/rdv24n1ab109 ◽

2012 ◽

Vol 24 (1) ◽

pp. 167

Author(s):

A. Shirasawa ◽

Y. Nakamura ◽

A. Ideta ◽

Y. Oono ◽

M. Urakawa ◽

...

Keyword(s):

Corpus Luteum ◽

Pregnancy Rate ◽

Embryo Transfer ◽

Blood Clot ◽

Pregnancy Rates ◽

Bovine Embryo ◽

Tissue Area ◽

The Mean ◽

Rectal Palpation ◽

Experimental Group

Recipient animals for bovine embryo transfer (ET) are routinely selected according to the morphology of the corpus luteum (CL) estimated by rectal palpation. However, rectal palpation is not a precise method of diagnosing the functional status of a CL. Ovarian ultrasonography (US) may be used to improve such diagnoses. The aim of this study was to evaluate the relationship between ultrasonographic images of CL and pregnancy rates after ET in Holstein heifers to determine whether US can be used to select recipients for ET. Recipient heifers (n = 285) were selected by detection of natural oestrus or following oestrus synchronization using a progesterone-releasing intravaginal device (PRID; ASKA Pharmaceutical, Tokyo, Japan). Transrectal US was performed immediately before ET, on Days 6 to 8 of the oestrous cycle (oestrus = Day 0), using a B-mode scanner (HS1500V; Honda Electronics Co. LTD, Aichi, Japan) equipped with a 7.5-MHz linear-array transducer designed for intrarectal placement. A cross-sectional image of the maximal area of the CL and luteal cavity was obtained. The areas of the CL and luteal cavity were each calculated using the formula for the area of an ellipse (height/2 × width/2 × π). (1) Ultrasonic morphology of CL was classified into 3 types: without cavity (n = 128), with cavity (n = 145) and with blood clot (n = 12). (2) The luteal cavity was categorized into 3 groups: small (<100 mm2, n = 93), medium (100 ≤ x < 200 mm2, n = 32) and large (≥200 mm2, n = 20). (3) Luteinized tissue area (total area of CL minus the area of the luteal cavity) was categorized into 3 groups: small (<250 mm2, n = 61), medium (250 ≤ x < 350 mm2, n = 128) and large (≥350 mm2, n = 84). In vivo–produced embryos were transferred nonsurgically into the uterine horn ipsilateral to the CL. Pregnancy was determined by transrectal US on Days 30 to 40 of gestation. The pregnancy rates of each experimental group were analysed by logistic regression. In this study, the pregnancy rate did not differ significantly in each experimental group: (1) without cavity: 77.3% (99/128), with cavity: 75.2% (109/145) and blood clot: 75.0% (9/12); (2) small cavity: 73.1% (68/93), medium: 75.0% (24/32) and large: 85.0% (17/20). The mean area of the cavity was 100.8 ± 110.3 mm2 (mean ± standard deviation) and recipients with 0 to 539.7 mm2 sized cavities had successful pregnancies (observational range was 0 to 539.7 mm2). (3) The pregnancy rates of recipients that had small, medium and large luteinized tissue were 77.0% (47/61), 75.0% (96/128) and 77.4% (65/84), respectively. The mean area of luteinized tissue was 318.9 ± 90.3 mm2 and 155.0 to 620.0 mm2 sized luteinized tissue had pregnancy success (observational range was 132.8 to 620.0 mm2). In conclusion, the results from this study indicate that the presence of a luteal cavity or blood clot has no detrimental effect on pregnancy success after ET in Holstein heifers. Furthermore, no relationship was found between luteinized tissue area at the time of ET and pregnancy rate.

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162 FERTILITY OF BEEF RECIPIENTS FOLLOWING A FIXED-TIME EMBRYO TRANSFER PROTOCOL WHICH INCLUDED FOLLICLE STIMULATING HORMONE DILUTED IN HYALURONAN

Reproduction Fertility and Development ◽

10.1071/rdv25n1ab162 ◽

2013 ◽

Vol 25 (1) ◽

pp. 229

Author(s):

J. W. Thorne ◽

C. R. Looney ◽

J. F. Hasler ◽

D. K. Hockley ◽

D. W. Forrest

Keyword(s):

Corpus Luteum ◽

Pregnancy Rate ◽

Embryo Transfer ◽

Transfer Rate ◽

Animal Health ◽

Fixed Time ◽

Control Groups ◽

Pfizer Animal Health ◽

And Control ◽

Estradiol 17Β

This study was performed to test the viability of administering Folltropin-V® (FSH, Bioniche Animal Health) diluted in hyaluronan (MAP-5 50 mg, sodium hyaluronate, Bioniche Animal Health) to beef cows enrolled in a recipient synchronization protocol to evaluate its effect on recipient fertility. All recipients were administered an estradiol 17β (2.5 mg, IM) and progesterone (50 mg, IM) combination injection on Day 0, a CIDR® (progesterone 1.34 g, Pfizer Animal Health, Groton, CT, USA) was inserted for 7 days. Lutalyse® (dinoprost tromethamine, Pfizer Animal Health, 25 mg, IM) was administered at the time of CIDR removal on Day 7, and estradiol 17β (1 mg, IM) was administered on Day 8. On Day 16, the presence of at least one corpus luteum, detected via ultrasound, resulted in the recipient receiving an embryo (both fresh and frozen–thawed embryos were used). Embryos were not transferred into cows that did not show ultrasonic evidence of a CL. Dependent variables for which data were collected included circulating progesterone levels at the time of transfer and CL diameter, area, and circumference; measured in millimeters. The total study (n = 274) consisted of both wet (n = 85) and dry (n = 189) cows and included both Bos indicus (Brahman-influenced) crossbred (n = 93) and Bos taurus (Angus-based) cows (n = 181). The experiment consisted of cows being placed in either the treated or control groups, with treated cows receiving a single 40 mg (1 mL) IM injection of Folltropin-V in hyaluronan on Day 5 and control cows receiving no additional injections. Results are shown in Table 1. Transfer rate, conception rate, and pregnancy rate were tested for significance with chi-square analysis and remaining statistics were analyzed with a t-test: two-sample assuming equal variances. There were no significant differences found between the treated and control groups for transfer rate, conception rate, or pregnancy rate. Corpus luteum diameter was shown to be larger in control cows (P < 0.05); however, CL area and circumference did not differ significantly. Folltropin-V given with hyaluronan at a 40-mg dose on Day 5 did not improve fertility, induce a larger CL, or increase circulating progesterone levels in synchronized beef recipients as hypothesized. Further work is needed with Folltropin-V in hyaluronan to determine if an alternative dose or timing of administration would be more appropriate for improving fertility in recipients. Table 1.Fertility data in beef recipients following synchronization for fixed-time embryo transfer with a protocol that included (Treated) or did not include (Control) FSH in hyaluronan

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Dynamics of trophoblast differentiation in peri-implantation–stage human embryos

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1911362116 ◽

2019 ◽

Vol 116 (45) ◽

pp. 22635-22644 ◽

Cited By ~ 4

Author(s):

Rachel C. West ◽

Hao Ming ◽

Deirdre M. Logsdon ◽

Jiangwen Sun ◽

Sandeep K. Rajput ◽

...

Keyword(s):

Cell Types ◽

Migration And Invasion ◽

Human Embryos ◽

Main Body ◽

Hormone Production ◽

Embryonic Tissues ◽

Maternal Immune System ◽

Placental Hormone ◽

First Contact ◽

Implantation Period

Single-cell RNA sequencing of cells from cultured human blastocysts has enabled us to define the transcriptomic landscape of placental trophoblast (TB) that surrounds the epiblast and associated embryonic tissues during the enigmatic day 8 (D8) to D12 peri-implantation period before the villous placenta forms. We analyzed the transcriptomes of 3 early placental cell types, cytoTB (CTB), syncytioTB (STB), and migratoryTB (MTB), picked manually from cultured embryos dissociated with trypsin and were able to follow sublineages that emerged from proliferating CTB at the periphery of the conceptus. A unique form of CTB with some features of STB was detectable at D8, while mature STB was at its zenith at D10. A form of MTB with a mixed MTB/CTB phenotype arose around D10. By D12, STB generation was in decline, CTB had entered a new phase of proliferation, and mature MTB cells had begun to move from the main body of the conceptus. Notably, the MTB transcriptome at D12 indicated enrichment of transcripts associated with IFN signaling, migration, and invasion and up-regulation of HLA-C, HLA-E, and HLA-G. The STB, which is distinct from the STB of later villous STB, had a phenotype consistent with intense protein export and placental hormone production, as well as migration and invasion. The studies show that TB associated with human embryos is in rapid developmental flux during peri-implantation period when it must invade, signal robustly to the mother to ensure that the pregnancy continues, and make first contact with the maternal immune system.

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