Contralateral Uterine Horn Research Articles

Possible roles of bone morphogenetic protein 4 in regulating endometrial function in cows.

This study investigated the expression dynamics of bone morphogenetic protein 4 (BMP4) and its receptors (BMPR1A, BMPR1B, and BMPR2) in bovine endometrium and examined the physiological function and regulatory mechanism of BMP4 expression. The messenger RNA (mRNA) expression of BMP4 and its receptors was detected in bovine endometrium of both ipsilateral (corpus luteum [CL]-side) and contralateral (non-CL-side) uterine horns during the estrous cycle and early pregnancy. BMP4 protein levels were higher in the endometrial tissues obtained from those cows in early pregnancy than in the estrous cycle. Immunohistochemical analysis showed that BMP4 and its receptors were localized in endometrial epithelial cells. The addition of BMP4 to cultured endometrial epithelial cells did not affect caspase-3/-8 mRNA expression, whereas it significantly inhibited cell proliferation. Both prostaglandin (PG) E2 and PGF2α concentrations in the culture supernatant were decreased when stimulated by BMP4. Furthermore, BMP4 mRNA expression was increased by stimulation with tumor necrosis factor-α (TNF) and interferon-γ (IFNG). In conclusion, BMP4 is produced in bovine endometrial epithelial cells and may contribute to the regulation of cell proliferation and suppression of PG secretion through autocrine or paracrine mechanisms. BMP4 expression in the bovine endometrium may be regulated by TNF and IFNG.

The efficiency of obtaining twins by replanting embryos in preinseminated animals

The purpose of our work was to determine the effectiveness of using of dairy cows as recipients for obtaining twin calves of the meat direction of produc-tivity. The study was conducted in LLC "Agricultural enterprise" Nikolaevskoe", Lugovets village, Mglinsky district. For the experiment cows and first-calf heifers (n=50) were selected and divided into three groups according to the breed princi-ple. In group I (n=22) Swiss breed animals were selected, in group II (n=13) - black-and-white cows, in group III (n=15) - crossbred cattle. All animals selected for the experiment were previously subjected to hormonal stimulation in order to synchronize the sexual cycles. The synchronization scheme included the injection of GnRh analogues at a dose of 10 ml IM and PgF2a cloprostenol at a dose of 500 mg per animal. With the manifestation of symptoms of estrus, all animals were arti-ficially inseminated with the sexed semen of a Belgian blue bull. On the seventh day of the sexual cycle, all inseminated animals had the procedure of replanting a frozen-thawed embryo into the contralateral uterine horn relative to the corpus lu-teum diagnosed on the ovary. To avoid freemartinism of heifers, the recipients were implanted with sexed embryos which were previously obtained from donor cows of the Hereford breed. During the study, it was found that the recipients in groups I, II and III had twins in 45.46% (10/22), 15.39% (2/13) and 20.0% (3/15) cases of joint application of biotechnological methods. The birth of one transplant calf was recorded in group I in 18.18% (4/22) of cows, in group II - 23.08% (3/13) and in group III - 33.33% (5/15) respectively. The birth of one calf obtained from artificial insemination was noted in group I in 31.82% (7/22) of animals, in group II - 38.46% (5/13) and in group III - 40.0% (6/15) respectively. The rest of the cows had no calves. Thus, it was found that the percentage of twins in the progeny was significantly higher (P?0.01) in group I compared with groups II and III. In groups I, II and III the total yield of calves was 140.92% (31/22), 92.32% (12/13) and 113.33% (17/15), respectively.

A Rabbit Model for Optimization of Amniotic Fluid Components in the EXTrauterine Environment for Newborn Development (EXTEND) System

In this model article, we present a protocol for continuous amniotic fluid exchange in rabbits using a novel system to test the effects of growth factor-deficient, artificial amniotic fluid on bowel development. Background: Ideally, the EXTrauterine Environment for Neonatal Development (EXTEND) will provide physiologic support to the extreme premature infant. An important component of that environment is the amniotic fluid. Thus, we developed an animal model to study the growth factors found within amniotic fluid and inform design of a synthetic fluid to optimize fetal development. Methods: We designed a model of amniotic fluid exchange within the pregnant rabbit, continuously removing the natural fluid from around 2 fetuses per doe and replacing it with a physiologic electrolyte solution during the final 100 h of gestation. Two fetuses from the contralateral uterine horn were used as sham-operated controls. Thirty-eight fetuses were analyzed, 19 in each group. We analyzed the fetal growth and bowel development. Results: Ultrasound after 100 h of exchange showed equivalent fluid volumes, p = 0.63. Cultures were negative for bacterial colonization. Final fluid protein concentrations were 11.6% that of control fluid (mean 1,451 ± 224.2 vs. 12,491 ± 849.2 μg/mL). There was no significant difference in fetal growth, with experimental weights 91.4% of control weights, p = 0.07. Fetal bowel weights (90.1%, p = 0.16) and lengths (94.2%, p = 0.49) were also not significantly less compared to controls. There was no significant difference in villous height or crypt depth measurements between the groups, and absorptive capacity of the bowel was not different between groups, p = 0.44. Conclusion: This animal model allows for manipulation of the components of amniotic fluid. Marked reduction of natural amniotic fluid proteins during gestation does not appear to significantly impair fetal growth or bowel development. Further work with this model will assess the importance of amniotic fluid components for normal development to inform design of a synthetic fluid for use during EXTEND.

Effect of the side of ovulation on the uterine morphometry, blood flow, progesterone, oestradiol and nitric oxide during spontaneous and induced oestrus in lactating dairy cows

This study investigates the changes in endometrial thickness between ipsilateral and contralateral uterine horns, diameter, blood flow area and hormonal concentrations in cows during natural and induced oestrus. In the induced group, six cows received a controlled internal device insert for 7days. In the normal and induced oestrous group (n=12), Doppler was performed day after day from day 5 till day 15. Oestradiol, progesterone and nitric oxide metabolites (NOMs) were measured. Results showed that dorsal, ventral and cross-section diameter of the ipsilateral horn increased during the induced oestrous group to reach a maximum on day 2 than during normal one. The total coloured area in both horns in a normal and induced group was also affected (p<.001), as the total coloured area in the ipsilateral horns, increased in the induced group, reach a maximum on day 2 after ovulation, while in normal it reaches a maximum on day of ovulation. Oestradiol increased during the induced cycle than during the normal one, but progesterone increased during days after ovulation reaches a higher value on day 10 in both groups. Nitric oxide showed two peaks, one on the day 3 and another peak on day 5. Conclusion: The ipsilateral uterine horn different diameters and thickness increased more rapidly in the induced oestrous group than the natural oestrus group. A complete analysis of variations in P4 and E2 concentrations and endometrial thickness suggested that decreases in P4 concentrations were related to an increase in endometrial thickness and that increases in E2 concentrations enhanced these endometrial changes.

Technical note: Relationship between placentome location and gene expression in bovine pregnancy.

A novel, non-terminal surgical procedure to remove a single placentome from the pregnant ewe for gene expression and histological analyses was recently developed in our laboratory. This technique allows for evaluation of nutritional insults on placental development at more than one stage of gestation using a single animal. Early attempts to develop a similar technique in cattle were met with complications due to inaccessibility of the gravid uterine horn because of its location and mass. One alternative is to collect a placentome from the contralateral uterine horn; however, the question remains as to whether gene expression varies among placentomes based on location relative to the fetus. Pregnant heifers were maintained on forage during early gestation and later moved into pens with a Calan gate system (American Calan, Northwood, NH). On gestational day (GD) 158, five heifers were assigned to receive a hay-based diet formulated to meet 100% of NRC requirements, and five heifers were fed 70% of NRC requirements until necropsy on GD244. At necropsy, a single representative placentome was selected for analysis from the antimesometrial side: (1) of the gravid uterine horn central to the amnion, (2) over the allantois immediately adjacent to the amnion, (3) in the tip of the gravid uterine horn, and (4) in the tip of the contralateral uterine horn. Mean placentome weight was greater (P < 0.05) for locations central to the amnion and allantois compared to locations within the tips of the ipsilateral and contralateral horns, respectively. Gene expression for angiogenic factors (FGF2, ODC1, VEGFA, and FLT1), nutrient transporters (SLC7A1 and SLC2A1), and factors associated with hormone action (ESR1, IGF1, IGFBP3, CSH1, and PAG1) were unaffected (P > 0.05) by dietary treatment or location of the placentome. Results indicate that location of the placentome in relation to the fetus does not impact gene expression, enhancing the efficacy of nonterminal methodologies for sampling gene expression in placentomes.

Location relative to the corpus luteum affects bovine endometrial response to a conceptus.

In cattle, embryo transfer into the uterine horn contralateral to the corpus luteum results in a higher incidence of pregnancy loss compared to transfer into the ipsilateral horn. We have previously reported temporal changes in the endometrial transcriptome during the estrous cycle which differ between uterine horns. The objective of this study was to compare the transcriptomic response of endometrium from the ipsilateral and contralateral horns to an elongating conceptus. Cross-bred beef heifers (n = 16) were synchronized and either used to generate day 14 conceptuses following the transfer of in vitro-produced blastocysts or to obtain day 14 endometrial explants. Conceptuses were recovered on day 14 by post-mortem uterine flushing, placed individually on top of explants collected from the ipsilateral (IPSI-D14) or the contralateral (CONTRA-D14) uterine horn of cyclic heifers, and co-cultured for 6 h. The response to a conceptus was markedly different between uterine horns, with 61 and 239 differentially expressed genes (DEGs; false discovery rate <0.05) in the ipsilateral and contralateral horns, respectively, compared to their controls. Direct comparison between IPSI-D1 and CONTRA-D14 revealed 32 DEGs, including CXCL11, CXCL10, IFIT2, RSAD2 and SAMD9. Gene Ontology analysis of these 32 genes revealed ten enriched biological processes, mainly related to immune response and response to an external stimulus. These data indicate that the endometrial response to the presence of a conceptus varies between uterine horns in the same uterus and may contribute to the higher incidence of pregnancy loss following embryo transfer to the contralateral horn.

Do differences in the endometrial transcriptome between uterine horns ipsilateral and contralateral to the corpus luteum influence conceptus growth to day 14 in cattle?

Embryo transfer to the uterine horn contralateral to the ovary containing the corpus luteum (CL) negatively impacts pregnancy establishment in cattle. Our aim was to compare the transcriptome and ability of the ipsilateral and contralateral uterine horns to support preimplantation conceptus survival and growth to day 14. In experiment 1, endometrial samples from both horns were collected from synchronized heifers slaughtered on day 5, 7, 13, or 16 post-estrus (n=5 per time) and subjected to RNA sequencing. In experiment 2, 10 day 7 in vitro produced blastocysts were transferred into the uterine horn ipsilateral (n=9) or contralateral to the CL (n=8) or into both horns (i.e., bilateral, n=9) of synchronized recipient heifers. Reproductive tracts were recovered at slaughter on day 14, and the number and dimensions of recovered conceptuses were recorded for each horn. A total of 217, 54, 14, and 18 differentially expressed genes (>2-fold change, FDR P<0.05) were detected between ipsilateral and contralateral horns on days 5, 7, 13, and 16, respectively, with signaling pathways regulating pluripotency of stem cells, ErbB signaling pathway, and mTOR signaling pathway amongst the top canonical pathways. Site of embryo transfer did not affect recovery rate (48.0%, 168/350) or length of conceptuses (mean±SE 2.85±0.27 mm). Although differences in gene expression exist between the endometrium of uterine horns ipsilateral and contralateral to the CL in cattle, they do not impact conceptus survival or length between day 7 and 14.

Creation of a rabbit model for intrauterine adhesions using electrothermal injury.

The pathogenesis and therapeutic treatment of intrauterine adhesions (IUAs) remain unsolved, highlighting the need for stable and effective experimental animal models. In this study, uterine electrocoagulation of twenty-one female New Zealand White rabbits was carried out to establish an IUA model. As rabbits have two completely separate uterine horns, each rabbit had its own internal control: one uterine horn was given an electrothermal injury (Group A, n=21), and the contralateral uterine horn received no treatment and served as the control (Group B, n=21). The endometrial morphology, number of endometrial glands, area of endometrial fibrosis, and number of implanted fetuses were compared between the two groups. In Group A, the numbers of endometrial glands on Days 7 and 14 and the number of implanted fetuses were significantly lower than those in Group B (P<0.05, P<0.05, and P<0.01, respectively), while the ratio of the area with endometrial stromal fibrosis to the total endometrial area was significantly increased (P<0.01). These results suggest that this method of electrothermal injury is effective for the establishment of a rabbit IUA model between 7 and 14 d after surgery.

110 Temporal Changes in Endometrial Gene Expression Between Ipsi- and Contralateral Uterine Horns in Cattle

The transfer of an embryo into the uterine horn contralateral to the ovary bearing the corpus luteum has been associated with a decreased pregnancy rate in cattle compared with transfer into the ipsilateral horn. These findings suggest that the environment in the contralateral horn is less conducive to supporting conceptus development than that of the ipsilateral horn. Therefore, this study compared the endometrial transcriptome of the ipsi- and contralateral uterine horns during the luteal phase. Endometrial samples from the ipsi- (IPSI) and contralateral (CONTRA) horns were collected from synchronized nonpregnant beef heifers on Days 5, 7, 13 or 16 post-oestrus (n = 5 heifers per time point). Total RNA was isolated and sequenced. Differences in the transcriptome were determined by edgeR-robust analysis. Principal component analysis found that IPSI and CONTRA have distinct patterns of gene expression on each day, with Day 5 exhibiting the most variation and Day 16 being least variable. Further, the 2 uterine horns had distinct expression patterns on Day 5, with IPSI exhibiting significantly higher variation in gene expression compared twitho CONTRA. EdgeR-robust analysis found 217 (201 up- and 16 down-regulated), 54 (44 up- and 10 down-regulated), 14 (13 up- and 1 down-regulated), and 18 (14 up- and 4 down-regulated) differentially expressed genes (DEG; &gt;2-fold change, false discovery rate P &lt; 0.05) between IPSI and CONTRA endometria on Days 5, 7, 13, and 16 of the oestrous cycle, respectively. The top 5 canonical pathways associated with DEG between IPSI and CONTRA during the luteal phase of the oestrous cycle were involved in signalling pathways regulating pluripotency of stem cells (73/138), progesterone-mediated oocyte maturation (55/89), endometrial cancer (31/51), ErbB signalling pathway (50/87), and mTOR signalling pathway (36/61). The impact of DEG on signalling pathways was assessed using a pathway perturbation algorithm called Signalling Pathway Impact Analysis (SPIA). This topology-based pathway analysis was conducted using the Bioconductor ToPAseq package (https://bioconductor.org/packages/release/bioc/html/ToPASeq.html) and revealed that signalling pathways regulating pluripotency of stem cells showed the highest perturbation score when IPSI was compared with CONTRA irrespective of day. Discovering and cataloguing which pathways are perturbed in each uterine horn throughout the oestrous cycle may contribute to our understanding of the mechanisms underlying early embryonic loss. Ths study was supported by Science Foundation Ireland (13/IA/1983) and the Irish Department of Agriculture, Food and The Marine (13S528).

Establishment of mouse endometrial injury model by curettage or coagulation

Objective: To establish mouse endometrial injury model by curettage or coagulation. Methods: Female ICR mice were randomly allocated into 2 groups: in curettage group, a blunt 20G needle was inserted in one uterine horn with 0.05 megapascals negative pressure; in coagulation group, one uterine horn was coagulated using a monopolar electric needle with 0.5 watts power. In both groups the contra-lateral uterine horn was used as control. The morphological changes and thickness of endometrium were evaluated 1 week after operation. The endometrial samples were taken on d4 of pregnancy, and the expressions of endometrial receptivity-related cytokines were examined. The number of implanted embryos on each side of uterus was calculated on d10 of pregnancy. Results: There was no difference in operation time between 2 groups. In both groups, the endometrial glands and stroma were significantly reduced, and the endometrial thickness was also significantly decreased on injury side compared to contra-lateral horn. However, local injury was more severe in coagulation group, uterine obliteration and hydrops were developed in 2 mice of coagulation group, and none in curettage group. The expressions of leukemia inhibitory factor (LIF) and oncostatin M (OSM) were significantly reduced on injured side in both groups compared to opposite side; however, the expression of LIF and OSM in curettage group was higher than that in coagulation group. The numbers of implanted embryos were decreased in both groups on injured side compared to opposite side, and fetal death was only observed in coagulation group. Conclusion: Both curettage and coagulation can make injury on mouse endometrium, impair endometrial receptivity and reduce fertility. Curettage can cause moderate injury, and coagulation may lead to more severe injury.

Differential gene expression profiling of endometrium during the mid-luteal phase of the estrous cycle between a repeat breeder (RB) and non-RB cows

BackgroundRepeat breeding directly affects reproductive efficiency in cattle due to an increase in services per conception and calving interval. This study aimed to investigate whether changes in endometrial gene expression profile are involved in repeat breeding in cows. Differential gene expression profiles of the endometrium were investigated during the mid-luteal phase of the estrous cycle between repeat breeder (RB) and non-RB cows using microarray analysis.MethodsThe caruncular (CAR) and intercaruncular (ICAR) endometrium of both ipsilateral and contralateral uterine horns to the corpus luteum were collected from RB (inseminated at least three times but not pregnant) and non-RB cows on Day 15 of the estrous cycle (4 cows/group). Global gene expression profiles of these endometrial samples were analyzed with a 15 K custom-made oligo-microarray for cattle. Immunohistochemistry was performed to investigate the cellular localization of proteins of three identified transcripts in the endometrium.ResultsMicroarray analysis revealed that 405 and 397 genes were differentially expressed in the CAR and ICAR of the ipsilateral uterine horn of RB, respectively when compared with non-RB cows. In the contralateral uterine horn, 443 and 257 differentially expressed genes were identified in the CAR and ICAR of RB, respectively when compared with non-RB cows. Gene ontology analysis revealed that genes involved in development and morphogenesis were mainly up-regulated in the CAR of RB cows. In the ICAR of both the ipsilateral and contralateral uterine horns, genes related to the metabolic process were predominantly enriched in the RB cows when compared with non-RB cows. In the analysis of the whole uterus (combining the data above four endometrial compartments), RB cows showed up-regulation of 37 genes including PRSS2, GSTA3 and PIPOX and down-regulation of 39 genes including CHGA, KRT35 and THBS4 when compared with non-RB cows. Immunohistochemistry revealed that CHGA, GSTA3 and PRSS2 proteins were localized in luminal and glandular epithelial cells and stroma of the endometrium.ConclusionThe present study showed that endometrial gene expression profiles are different between RB and non-RB cows. The identified candidate endometrial genes and functions in each endometrial compartment may contribute to bovine reproductive performance.

Prediction of blastocyst development and implantation potential in utero based on the third cleavage and compaction times in mouse pre-implantation embryos.

Cytokinesis and cell division during pre-implantation embryonic development occur as an orchestrated spatiotemporal program. Cleavage, compaction, and blastulation in pre-implantation embryos are essential for successful implantation and pregnancy. Their alteration is associated with chromosomal imbalance and loss of developmental competence. In this study, we evaluated the time of cleavage and compaction as predictors for in vitro pre- and peri-implantation development and in utero implantation potential by time-lapse monitoring. Mouse 2-cell embryos were collected on 1.5 days post coitum (dpc) and were individually cultured to the outgrowth (OG) stage (7.5 dpc). Developmental stages were classified as 3-cell, 4-cell, 8-cell, morula, blastocyst, and OG. Cut-off times for successful blastocyst development were determined by receiver operating characteristic curve analysis. When cut-off times were set as 9 h for the third cleavage from the 2- to 4-cell stage, and 40 h for compaction from the 2-cell to morula stage, blastocyst and OG development rates, respectively, were significantly higher (P < 0.0001). Embryos were grouped according to the above cut-off time and transferred to the contralateral uterine horn on 3.5 dpc. Implantation rates in utero on 5.5 dpc were significantly higher in early third cleaved (≤ 9 h from 2- to 4-cell) and early compacted embryos (≤ 40 h from 2-cell to morula) than those in delayed embryos (P < 0.05). Therefore, the time of the third cleavage from 2- to the 4-cell stage and compaction from 2-cell to morula stage may be a useful morphokinetic parameter for predicting developmental potential, including successful implantation and pregnancy in human in vitro fertilization-embryo transfer programs.

Assessment of Uterine Vascular Perfusion During the Estrous Cycle of Mares in Connection to Circulating Leptin and Nitric Oxide Concentrations

The objective of this study was to find difference in vascular perfusion of uterine horns or uterine body throughout the estrous cycle and their relation to circulating nitric oxide and leptin concentrations. Five cyclic mares were subjected to transrectal Doppler ultrasonography and blood sampling for 18 days. Area of color and power Doppler modes was measured in pixels. Day (P = .0001) of the estrous cycle and ovulation (P = .0001) influenced uterine blood flow. Uterine body blood flow directed away from the transducer (blue, P = .0001) increased from day −5 until day 0 (day of ovulation), and its power (P = .0001) blood flow increased from day −6 until day 0; then, both decreased until days 12 and 10, respectively. Conversely to the contralateral uterine horn, ipsilateral uterine horn blood flow directed away from the transducer (blue, P = .0001) increased from day −5 until day −1, and its power (P = .0001) blood flow increased from day −6 until day 0; then, both decreased until day 10. Nitric oxide concentrations (P = .0001) attained two major peaks; the first on day −3 and the other persisted from day 2 until day 5. Leptin concentrations increased (P > .001) with a maximum value on day 0 and then decreased until a minimum value on day 9. In conclusion, during the estrous cycle, ipsilateral uterine horn and uterine body blood vessels had similar blood flow. Both leptin and nitric oxide played a role during follicle growth, ovulation, and corpus luteum development and modulated uterine blood flow before and after ovulation.

Migração espermática em suínos após inseminação artificial intrauterina profunda

A inseminação artificial intrauterina profunda (IIP) é de grande importância para a indústria suinícola, em função do maior número de doses produzidas por reprodutores de alto mérito genético e da possibilidade da utilização de biotecnologias, como sêmen sexado e/ou congelado. Entretanto, necessita-se compreender com maior propriedade os mecanismos pelos quais os espermatozoides colonizam as tubas uterinas. Assim sendo, pretende-se com o presente experimento avaliar a existência ou não de migração intraperitoneal de espermatozoides inseminados profundamente em um dos cornos uterinos, mediante a obtenção de oócitos fertilizados no corno contralateral à inseminação e seccionado na base, na junção com o corpo do útero. Quatorze fêmeas pluríparas foram divididas em dois grupos experimentais, sendo que em um deles as fêmeas foram submetidas à secção da base de um dos cornos uterinos (Grupo Operado, n = 7), enquanto as do Grupo Controle (n = 7) não foram submetidas a nenhuma intervenção cirúrgica. Ambos os grupos foram submetidos à IIP, sendo as fêmeas abatidas 5±1,2 dias após a última inseminação. Os sistemas genitais das fêmeas foram coletados, dissecados e o número de corpos lúteos contados em ambos os ovários. A recuperação dos embriões foi feita por meio de lavagem das tubas e cornos uterinos com solução de PBS (Phosphate Buffered Saline), após o que se avaliou os fluidos coletados em lupa para a identificação de embriões. Em ambos os grupos experimentais, foram encontrados embriões nos segmentos do sistema genital de ambos os lados. Apenas uma fêmea apresentou embriões nos segmentos em somente um dos lados no grupo operado. Diante dos resultados aqui observados, concluiu-se que a migração espermática no suíno pode ocorrer tanto por via retrógrada pelo útero quanto por migração intraperitoneal. Estes achados certamente contribuirão para aumentar a eficiência da técnica de IIP, sendo de grande valia para o aprimoramento da indústria suinícola.

Composition of the bovine uterine proteome is associated with stage of cycle and concentration of systemic progesterone

Early embryonic loss accounts for over 70% of total embryonic and foetal loss in dairy cattle. Early embryonic development and survival is associated with the concentration of systemic progesterone. To determine if the uterine proteome is influenced by stage of cycle or systemic progesterone concentrations, uterine flushings were collected from the ipsi- and contralateral uterine horns of beef heifers on Days 7 (n = 10) and 15 (n = 10) of the oestrous cycle. Animals were separated into low or high progesterone groups based on plasma progesterone concentrations on Day 5 of the cycle. Samples were albumin depleted before iTRAQ R labeling and subsequent strong cation exchange-LC-MS/MS analyses. A total of 20 proteins were up to 5.9-fold higher (p<0.05) and 20 were up to 2.3-fold lower on Day 15 compared toDay 7. In addition, the expression of a number of proteins on Day 7 and/or 15 of the cycle was correlated with progesterone concentrations during Days 3–7 or the rate of change in progesterone between Days 3 and 7. This study highlights the dynamic changes occurring in the microenvironment surrounding the embryo during this period. The findings here also support the hypothesis that progesterone supports embryonic development by altering the maternal uterine environment.

Effect of Conceptus on Transforming Growth Factor (TGF) β1 mRNA Expression and Protein Concentration in the Porcine Endometrium— &lt;i&gt;In Vivo&lt;/i&gt; and &lt;i&gt;In Vitro&lt;/i&gt; Studies

Transforming growth factor (TGF) β and its receptors are expressed at the conceptus-maternal interface during early pregnancy in the pig. The present studies were conducted to examine: (1) the effect of conceptus products on TGFβ1 mRNA expression and protein concentration in the porcine endometrium using in vivo and in vitro models, and (2) the effect of TGFβ1 on proliferation of porcine trophoblast cells in vitro. During in vivo experiments, gilts with one surgically detached uterine horn were slaughtered on days 11 or 14 of the estrous cycle and pregnancy. For in vitro studies, endometrial explants and luminal epithelial (LE) cells co-cultured with stromal (ST) cells were treated with conceptus-exposed medium (CEM). Moreover, porcine trophoblast cells were treated with TGFβ1, and the number of viable cells was measured. On day 11, the presence of conceptuses had no effect on TGFβ1 mRNA expression, but decreased the TGFβ1 protein concentration in the connected uterine horn compared with the detached uterine horn. In contrast to day 11, on day 14 after estrus, TGFβ1 mRNA expression and protein content in the endometrium collected from the gravid uterine horn were greater when compared with the contralateral uterine horn. The treatment of endometrial slices with CEM resulted in greater TGFβ1 mRNA expression and protein secretion. LE cells responded to CEM with an increased TGFβ1 mRNA level. Moreover, TGFβ1 stimulated the proliferation of day 14 trophoblast cells. In summary, porcine conceptuses may regulate TGFβ1 synthesis in the endometrium at the time of implantation. TGFβ1, in turn, may promote conceptus development by increasing the proliferation of trophoblast cells.

Improved Fertilization Success Using Laparoscopic Oviductal Artificial Insemination with Low Sperm Numbers in Domestic Cats.

Artificial insemination (AI) is a valuable tool for infusing genetic variation into stagnant populations, especially using frozen-thawed spermatozoa, and for propagating physically or behaviorally incompatible individuals. Within zoological institutions, AI has received growing interest for genetic management of endangered felids, provided that AI success can be optimized for applied usage. Traditionally, laparoscopic AI in cats involves treatment with equine chorionic gonadotropin (eCG) followed 80-85h later by human CG (hCG) to induce follicular development and ovulation, with subsequent bilateral sperm deposition into the uterine lumen. However, hCG, a large glycoprotein, may remain in circulation for several days post-injection, generating undesirable secondary ovulations. Uterine AI also requires relatively high numbers of spermatozoa to achieve sperm transport through the uterotubal junction and fertilization within the oviduct. Furthermore, sperm recovery from male cats frequently is poor, limiting the number of spermatozoa available for AI. Alterations in the AI protocol, using short-acting porcine luteinizing hormone (pLH) as the ovulatory signal and oviductal AI for sperm deposition, could improve fertilization success while requiring fewer spermatozoa. Our objectives in this study were to assess pregnancy and fertilization success in cats treated with one of two gonadotropin regimens (eCG/hCG vs. eCG/pLH) and inseminated laparoscopically at two sperm deposition sites, the uterus (UT) and the contralateral oviduct (OV). Sixteen female domestic cats were randomly assigned to either eCG (100 IU)/hCG (75 IU) or eCG/pLH (1000 IU) treatment groups. All 16 females ovulated following gonadotropin treatment and were inseminated with low sperm numbers (1x106 motile sperm/site; 5 µl volume) in one uterine horn and one contralateral oviduct using freshly collected semen from a different male for each site. Semen samples were obtained from two males of proven fertility via artificial vagina. Pregnant females were spayed at 20-21 days post-AI and recovered fetuses assessed for paternity using genetic analysis. Comparing gonadotropin regimens, similar numbers of females became pregnant following eCG/hCG (75%, 6/8) versus eCG/pLH (63%, 5/8). The number of corpora lutea (CL) at AI was similar between regimens, but hCG treatment increased the number of CLs at day 20 post-AI. Although hCG and pLH treatments produced similar numbers of normal fetuses, implantation abnormalities (e.g. empty gestational sacs, malformed placentae) were observed in the hCG, but not pLH, group. Overall, 11 (of 16; 69%) females became pregnant (5 cats from OV AI only, 2 from UT AI only, and 4 from both sites). In comparing insemination sites, more fetuses resulted from OV AI (36/49; 73%) than UT AI (13/49; 27%). To assess capacity for term pregnancies, three additional females were treated with eCG/pLH and inseminated in one oviduct and the contralateral uterine horn. All three females became pregnant (2-3 fetuses each) and healthy kittens were produced. In summary, laparoscopic oviductal AI with low sperm numbers in eCG/pLH-treated females resulted in high pregnancy and fertilization percentages in domestic cats. These findings suggest that this technique may have value for propagating endangered nondomestic cat species. Our recent success in using oviductal AI in eCG/pLH-treated ocelots (Leopardus pardalis) to produce a healthy ocelot kitten supports this cross-species applicability. (platform)

V1713 PEDIATRIC ROBOT-ASSISTED LAPAROSCOPIC RESECTION OF A FUNCTIONAL NON-COMMUNICATING UTERINE HORN

You have accessJournal of UrologyPediatrics & Reconstruction1 Apr 2011V1713 PEDIATRIC ROBOT-ASSISTED LAPAROSCOPIC RESECTION OF A FUNCTIONAL NON-COMMUNICATING UTERINE HORN Manoj Rao, Andrew Marks, and Bruce Lindgren Manoj RaoManoj Rao Chicago, IL More articles by this author , Andrew MarksAndrew Marks Chicago, IL More articles by this author , and Bruce LindgrenBruce Lindgren Chicago, IL More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2011.02.2038AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES We present robot assisted laparoscopic resection of a non-communicating functional uterine horn in a pediatric patient. To our knowledge, this is the first reported robot assisted procedure for this indication in a pediatric patient. METHODS A 13-year-old female presented from an outside hospital with left abdominal pain. She was found to have a cystic pelvic mass and underwent open resection via a pfannensteil incision with pathology showing hematosalpinx and encased ovary. Post operatively, she returned with recurrent left abdominal pains and left leg numbness and tingling. MRI showed a right unicornuate uterus and a non-communicating, functional left uterine horn with hematometros as well as left sided renal agenesis. She was transferred to our facility for further care. A robotic resection of the non communicating uterine horn was performed. RESULTS After positioning in dorsal lithotomy, we performed examination under anesthesia, cystoscopy and vaginoscopy, identifying a single right-sided cervix and a mass effect on the left pushing on the bladder. We proceeded with robot assisted laparoscopic resection. We used an 8.5 mm camera port 2 cm superior to the umbilicus. We identified the left uterine horn and the normal contralateral uterine horn, ovary and fallopian tube. Two 8 mm robotic ports were placed in the right and left lower quadrants and a 10 mm assistant port was placed through the right lateral aspect of her prior pfannensteil incision. The distended uterine horn was punctured and 60 ml of hemolyzed blood aspirated for decompression. The sigmoid colon was aherent to the lateral aspect of the uterine horn due to previous salpingo-oophorectomy, and was dissected away. The vascular pedicle to the left uterine horn was ligated with a harmonic scalpel. The fibrotic attachment of the uterine horn to the vagina was identified and divided, with guidance from an assistant's finger to delineate the lateral wall of the vagina and the rectum. The specimen was placed into an endocatch bag inserted via the assistant port, and the incision extended along her prior Pfanensteil incision specimen retrieval. Estimated blood loss was 100 ml. She was discharged home post operative day three. On follow up she had no abdominal pain and lower extremity numbness had resolved. CONCLUSIONS Robot-assisted laparoscopic resection of a functional uterine horn can be performed successfully, safely, and with good outcome. This technique offers a minimally invasive approach to a complicated internal genital anomaly and is another robot assisted procedure for the urologist's armamentarium. © 2011 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 185Issue 4SApril 2011Page: e688 Advertisement Copyright & Permissions© 2011 by American Urological Association Education and Research, Inc.MetricsAuthor Information Manoj Rao Chicago, IL More articles by this author Andrew Marks Chicago, IL More articles by this author Bruce Lindgren Chicago, IL More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Direct ovarian - uterine transfer of progesterone increases embryo survival in gilts

This study employed a unilateral ovariectomy model to investigate the relevance of the local supply of progesterone (ovary) compared with the systemic supply of progesterone, in terms of embryo survival in the ipsilateral uterine horn as opposed to the contralateral uterine horn. Thirty gilts were unilaterally ovariectomised (ULO) during the luteal stage of their first oestrous cycle. Half of the ULO gilts were fed at 1.2 maintenance requirement (M), while the other half were fed at 2.4M. Across ULO gilts 0.8 more embryos survived in the ipsilateral horn compared with the contralateral horn at Day 35 of gestation (P<0.05). In ULO gilts on the 2.4M feed level the difference (+1.3; P<0.05) between the ipsi- and contralateral horn was more pronounced than on the 1.2M feed level (+0.4; NS). The higher feed level reduced circulating levels of systemic progesterone on Day 5 of pregnancy but not embryo survival at Day 35. However, post-implantation embryo survival was lower on the low feed level. In conclusion, these data indicate that local progesterone supply from the ovaries to the uterus contributes to the probability of embryo survival.

Effect of intra-uterine injection of H2 receptor blockers on implantation in albino Wistar rats

The role of histamine in various stages of female reproductive functions is now understood. The anti-implantation activity of various H2 receptor blockers had been established earlier by our laboratory. The present studies were conducted to evaluate the local effects of direct intra-uterine injection of H2 receptor blockers on implantation in albino Wistar rats. The individual administration of ranitidine, famotidine and roxatidine by direct intra-uterine injection on Day 4 of pregnancy in female albino Wistar rats produced 100% anti-fertility activity probably by acting on the H2 receptors on the blastocyst. Control group animals delivered normal litters. Unilateral injection of roxatidine inhibited implantation in the treated and contralateral uterine horns of rats indicating its systemic as well as local anti-implantation activity while ranitidine and famotidine showed only the local effects on the treated uterine horns suggesting the blockade of locally derived uterine histamine in implantation. Histopathological examination of uterus revealed the absence of morphological damage to the endometrial epithelium. It is concluded that, the intra-uterine application of H2 receptor blockers prevents conception in rats. Key words: Anti-implantation activity H2 receptor blockers, ranitidine, famotidine, roxatidine, intra-uterine injection, implantation.