-
Reproduction (Cambridge, England) May 2020We previously demonstrated that 5'-AMP-activated protein kinase (AMPK) is essential for normal reproductive functions in female mice. Conditional ablation of Prkaa1 and...
We previously demonstrated that 5'-AMP-activated protein kinase (AMPK) is essential for normal reproductive functions in female mice. Conditional ablation of Prkaa1 and Prkaa2, genes that encode the α1 and α2 catalytic domains of AMPK, resulted in early reproductive senescence, faulty artificial decidualization, uterine inflammation and fibrotic postparturient endometrial regeneration. We also noted a delay in the timing of embryo implantation in Prkaa1/2d/d female mice, suggesting a role for AMPK in establishing uterine receptivity. As outlined in new studies here, conditional uterine ablation of Prkaa1/2 led to an increase in ESR1 in the uteri of Prkaa1/2d/d mice, resulting in prolonged epithelial cell proliferation and retention of E2-induced gene expression (e.g. Msx1, Muc1, Ltf) through the implantation window. Within the stromal compartment, stromal cell proliferation was reduced by five-fold in Prkaa1/2d/d mice, and this was accompanied by a significant decrease in cell cycle regulatory genes and aberrant expression of decidualization marker genes such as Hand2, Bmp2, Fst and Inhbb. This phenotype is consistent with our prior study, demonstrating a failure of the Prkaa1/2d/d uterus to undergo decidualization. Despite these uterine defects, ovarian function seemed to be normal following ablation of Prkaa1/2 from peri-ovulatory follicles in which ovulation, luteinization and serum progesterone levels were not different on day 5 of pregnancy or pseudopregnancy between Prkaa1/2fl/fl and Prkaa1/2d/d mice. These cumulative findings demonstrate that AMPK activity plays a prominent role in mediating several steroid hormone-dependent events such as epithelial cell proliferation, uterine receptivity and decidualization as pregnancy is established.
Topics: AMP-Activated Protein Kinases; Animals; Cell Proliferation; Embryo Implantation; Epithelial Cells; Estradiol; Estrogen Receptor alpha; Female; Mice; Mice, Knockout; Stromal Cells; Uterus
PubMed: 32191914
DOI: 10.1530/REP-19-0402 -
Scientific Reports Feb 2020Embryo transfer has been used as one of the essential reproductive technologies for production of new strains and maintenance of genetic resources in animals. Mating...
Embryo transfer has been used as one of the essential reproductive technologies for production of new strains and maintenance of genetic resources in animals. Mating with vasectomised male rats is a requirement for inducing pseudopregnancy in female rats selected for embryo transfer. Although this procedure has been used routinely, large breeding space and high expenditure are required to maintain a sufficient number of females and vasectomised males. This study was performed to induce pseudopregnancy in females by artificial stimulation using sonic vibration instead of vasectomised males. The females continued to be in the dioestrus stage for at least 14 days after artificial stimulation was performed. Of fresh 2-cell embryos that transferred into the oviducts of females after artificial stimulation, 56% was implanted and 50% was developed to offspring. Approximately 46% of the frozen 2-cell embryos were implanted and 24% developed into offspring. Furthermore, 66% of the fresh pronuclear embryos were implanted and 60% developed into offspring. This study successfully induced pseudopregnancy in rat females by artificial stimulation using a sonic vibration. This method, 'Easy-ET', was useful for efficient production and maintenance of rat strains.
Topics: Animals; Breeding; Cryopreservation; Embryo Implantation; Embryo Transfer; Embryo, Mammalian; Estrus; Fallopian Tubes; Female; Male; Pregnancy; Pseudopregnancy; Rats; Sound; Vasectomy; Vibration
PubMed: 32066799
DOI: 10.1038/s41598-020-59611-1 -
Molecular Human Reproduction Mar 2020The study investigated the effect of normal and supraphysiological (resulting from gonadotropin-dependent ovarian stimulation) levels of estradiol (E2) and progesterone...
Uterine aquaporin expression is dynamically regulated by estradiol and progesterone and ovarian stimulation disrupts embryo implantation without affecting luminal closure.
The study investigated the effect of normal and supraphysiological (resulting from gonadotropin-dependent ovarian stimulation) levels of estradiol (E2) and progesterone (P4) on mouse uterine aquaporin gene/protein (Aqp/AQP) expression on Day 1 (D1) and D4 of pregnancy. The study also examined the effect of ovarian stimulation on uterine luminal closure and uterine receptivity on D4 of pregnancy and embryo implantation on D5 and D7 of pregnancy. These analyses revealed that the expression of Aqp3, Aqp4, Aqp5 and Aqp8 is induced by E2 while the expression of Aqp1 and Aqp11 is induced by P4. Additionally, P4 inhibits E2 induction of Aqp3 and Aqp4 expression while E2 inhibits Aqp1 and Aqp11 expression. Aqp9, however, is constitutively expressed. Ovarian stimulation disrupts Aqp3, Aqp5 and Aqp8 expression on D4 and AQP1, AQP3 and AQP5 spatial expression on both D1 and D4, strikingly so in the myometrium. Interestingly, while ovarian stimulation has no overt effect on luminal closure and uterine receptivity, it reduces implantation events, likely through a disruption in myometrial activity and embryo development. The wider implication of this study is that ovarian stimulation, which results in supraphysiological levels of E2 and P4 and changes (depending on the degree of stimulation) in the E2:P4 ratio, triggers abnormal expression of uterine AQP during pregnancy, and this is associated with implantation failure. These findings lead us to recognize that abnormal expression would also occur under any pathological state (such as endometriosis) that is associated with changes in the normal E2:P4 ratio. Thus, infertility among these patients might in part be linked to abnormal uterine AQP expression.
Topics: Animals; Aquaporins; Embryo Implantation; Embryo Transfer; Estradiol; Female; Gene Expression Regulation, Developmental; Mice; Mice, Inbred C57BL; Mifepristone; Ovulation Induction; Pregnancy; Progesterone; Pseudopregnancy; Uterus; Water
PubMed: 31977023
DOI: 10.1093/molehr/gaaa007 -
Animals : An Open Access Journal From... Sep 2019Aggressiveness is one of the main problems in group housing of rabbit does. The aim of the experiment was to investigate the frequency of aggressiveness and mating...
Aggressiveness is one of the main problems in group housing of rabbit does. The aim of the experiment was to investigate the frequency of aggressiveness and mating behaviour as well as the lifespan of does depending on group composition. The female and male rabbits were housed in one of the 7.7 m pens (four females and one male per pen). Based on the ages of female rabbits two homogenous groups (HOM) were formed containing four 17-week-old females and two heterogeneous groups (HET) containing three 17-week-old and one 1-year-old female. Twenty-four-hour video recordings were taken during the first month after assembling the groups, and the aggressive actions (fights) and matings were counted. The lifespan was examined over a 200-day experimental period. On the day after assembling the groups the number of fights among does was high in HET group. The same aggressive behaviour only started a week later in HOM group, and some fights between females and the male were also observed. The daily peaks of aggressiveness were in the morning (after the light on) and in the evening (before and after the lights off). The primary position of females in the hierarchy was clear but sometimes no differences were detected among the subordinate females. The mortality of does was connected with their rank order. The number of matings was very high on the day of assembling the groups and a second small peak was observed at the end of the hypothetical pseudo-pregnancy. In addition to mating between male and females, female-female and female-male mounting was also observed. Despite of the small sample size it seems that aggressive behaviour is frequent in group housing systems, which is contrary to animal welfare. Natural mating is not effective in group-housing system.
PubMed: 31547162
DOI: 10.3390/ani9100708 -
Scientific Reports Sep 2019Female giant pandas show complex reproductive traits, being seasonally monoestrus, displaying a variable length embryonic diapause and exhibiting pseudopregnancy....
Female giant pandas show complex reproductive traits, being seasonally monoestrus, displaying a variable length embryonic diapause and exhibiting pseudopregnancy. Currently, there is no confirmatory non-invasive biomarker of blastocyst implantation or pregnancy. This study aimed to monitor urinary estrogens across gestation in pregnancy (n = 4), pseudopregnancy (n = 4) and non-birth cycles (n = 5) in the giant panda. A pregnancy-specific profile of estrogens corrected for urinary specific gravity was identified during the gestation period. Pregnant females showed increasing concentrations of estrogens for 29 days until birth, no increase was observed during pseudopregnancy and the two profiles were distinguishable from each other for the final 2 weeks of the cycle suggesting the estrogens are of placental origin. This allowed a nomogram, starting at a known fixed point during the cycle, to be created and tested with cycles of known outcome, and cycles which were inseminated but did not result in a birth. Non-birth profiles showed deviations from that of pregnancy. We believe these deviations indicate the point of failure of the placenta to support a developing cub. Non-invasive longitudinal monitoring of estrogen concentrations therefore has the potential to be developed as a panda pregnancy test to predict viable cub development.
Topics: Animals; Biomarkers; Estrogens; Female; Pregnancy; Ursidae
PubMed: 31484972
DOI: 10.1038/s41598-019-49288-6 -
PloS One 2019Recent studies indicate that FoxO1 has roles in female reproductive system, especially in maternal endometrium. Although various cellular aspects and molecular pathways...
Recent studies indicate that FoxO1 has roles in female reproductive system, especially in maternal endometrium. Although various cellular aspects and molecular pathways have been identified, the exact molecular characteristics of embryo implantation are still not completely understood. In this study, we aimed to investigate uterine expression and regulation of FoxO1 during peri-implantation period in mice. Experimental mouse models including, normal pregnancy, pseudopregnancy, artificial decidualization, and delayed implantation and activation were performed. Our results showed that FoxO1 expression was spatiotemporal in mouse endometrial tissue throughout peri-implantation period and its expression was significantly upregulated in luminal and glandular epithelium at the time of implantation. Moreover, on day 5 morning (09:00 AM) of pregnancy, expression of FoxO1 was cytoplasmic in endometrial luminal epithelial cells where embryo homing takes place. With progressing time on day 5 evening (19:00 PM) of pregnancy FoxO1 expression was nuclear in luminal epithelium at implantation site. Pseudopregnancy and artificial decidualization models indicated that FoxO1 expression was regulated by pregnancy hormones. Delayed implantation and activation model indicated that FoxO1 expression at the time of implantation is dependent upon activation status of blastocyst due to E2 induction and uterine sensitivity to implantation. In conclusion, our findings highlight a perspective for FoxO1 expression and regulation in mouse uterus during peri-implantation period indicating that its expression is regulated by implanting embryo and pregnancy hormones.
Topics: Animals; Blastocyst; Decidua; Embryo Implantation, Delayed; Female; Forkhead Box Protein O1; Gene Expression Regulation; Mice; Mice, Inbred BALB C; Pregnancy; Pseudopregnancy
PubMed: 31120913
DOI: 10.1371/journal.pone.0216814 -
Kisspeptin/kisspeptin receptor system in pseudopregnant rabbit corpora lutea: presence and function.Scientific Reports Mar 2019Kisspeptin (KiSS) and its related receptors (KiSS1R) have a critical role in the reproduction of mammals. The KiSS/KiSS1R system is expressed in numerous reproductive...
Kisspeptin (KiSS) and its related receptors (KiSS1R) have a critical role in the reproduction of mammals. The KiSS/KiSS1R system is expressed in numerous reproductive organs including the ovary. Here, we studied the expression of the KiSS/KiSS1R system and its functional role in rabbit corpora lutea (CL) at days 4 (early-), 9 (mid-), and 13 (late-stage) of pseudopregnancy. In vitro progesterone, prostaglandin (PG) F2α (PGF2α) and E2 (PGE2) productions and prostaglandin-endoperoxide synthase 1 (PTGS1) and 2 (PTGS2) activities were evaluated. Immune reactivity (IR) for KiSS and KiSS1R were detected in luteal cells at nuclear and cytoplasmic level at all luteal stage for KiSS and only at early- and mid-stage for KiSS1R; IR decreased from early- to later stages of pseudopregnancy. The KiSS-10 augmented progesterone and PGE2 and diminished PGF2α secretions by early- and mid-CL; KiSS-10 reduced PTGS2 activity at early- and mid-stages, but did not affect PTGS1 at any luteal stages. The antagonist KiSS-234 counteracted all KiSS-10 effects. This study shows that the KiSS/KiSS1R system is expressed in CL of pseudopregnant rabbits and exerts a luteotropic action by down-regulating PTGS2, which decreases PGF2α and increases PGE2 and progesterone.
Topics: Animals; Cyclooxygenase 2; Dinoprost; Dinoprostone; Female; Kisspeptins; Luteal Cells; Pregnancy; Pseudopregnancy; Rabbits; Receptors, Kisspeptin-1
PubMed: 30911071
DOI: 10.1038/s41598-019-41623-1 -
Scientific Reports Feb 2019Successful pregnancy requires the establishment of a complex dialogue between the implanting embryo and the endometrium. Knowledge regarding molecular candidates...
Successful pregnancy requires the establishment of a complex dialogue between the implanting embryo and the endometrium. Knowledge regarding molecular candidates involved in this early communication process is inadequate due to limited access to primary human endometrial epithelial cells (EEC). Since pseudo-pregnancy in rodents can be induced by mechanical scratching of an appropriately primed uterus, this study aimed to investigate the expression of mechanosensitive ion channels in EEC. Poking of EEC provoked a robust calcium influx and induced an increase in current densities, which could be blocked by an inhibitor of mechanosensitive ion channels. Interestingly, RNA expression studies showed high expression of PIEZO1 in EEC of mouse and human. Additional analysis provided further evidence for the functional expression of PIEZO1 since stimulation with Yoda1, a chemical agonist of PIEZO1, induced increases in intracellular calcium concentrations and current densities in EEC. Moreover, the ion channel profile of human endometrial organoids (EMO) was validated as a representative model for endometrial epithelial cells. Mechanical and chemical stimulation of EMO induced strong calcium responses supporting the hypothesis of mechanosensitive ion channel expression in endometrial epithelial cells. In conclusion, EEC and EMO functionally express the mechanosensitive PIEZO1 channel that could act as a potential target for the development of novel treatments to further improve successful implantation processes.
Topics: Animals; Endometrium; Epithelial Cells; Female; Humans; Ion Channels; Mice; Organoids
PubMed: 30741991
DOI: 10.1038/s41598-018-38376-8 -
Scientific Reports Sep 2018A complex analysis of acute kidney injury (AKI) in pregnant women shows that it is caused by the interaction of gestation-associated pathologies and beneficial signaling...
A complex analysis of acute kidney injury (AKI) in pregnant women shows that it is caused by the interaction of gestation-associated pathologies and beneficial signaling pathways activated by pregnancy. Studies report an increase in the regeneration of some organs during pregnancy. However, the kidney response to the injury during pregnancy has not been addressed. We investigated the mechanisms of the pregnancy influence on AKI. During pregnancy, the kidneys were shown to be more tolerant to AKI. Pregnant animals showed remarkable preservation of kidney functions after ischemia/reperfusion (I/R) indicated by the decrease of serum creatinine levels. The pregnant rats also demonstrated a significant decrease in kidney injury markers and an increase in protective markers. Two months after the I/R, group of pregnant animals had a decreased level of fibrosis in the kidney tissue. These effects are likely linked to increased cell proliferation after injury: using real-time cell proliferation monitoring we demonstrated that after ischemic injury, cells isolated from pregnant animal kidneys had higher proliferation potential vs. control animals; it was also supported by an increase of proliferation marker PCNA levels in kidneys of pregnant animals. We suggest that these effects are associated with hormonal changes in the maternal organism, since hormonal pseudopregnancy simulated effects of pregnancy.
Topics: Acute Kidney Injury; Animals; Apoptosis; Cell Proliferation; Cells, Cultured; Female; Humans; Kidney; Pregnancy; Protective Factors; Rats; Reperfusion Injury
PubMed: 30266919
DOI: 10.1038/s41598-018-32801-8 -
Veterinary World Apr 2018This study was conducted to estimate the prevalence of pseudopregnancy in goats and to investigate potential risk factors associated with the condition in Khartoum State.
AIM
This study was conducted to estimate the prevalence of pseudopregnancy in goats and to investigate potential risk factors associated with the condition in Khartoum State.
MATERIALS AND METHODS
A cross-sectional study was carried out from March 2015 to February 2016. A total of 378 female goats which presented to the Veterinary Teaching Hospital, College of Veterinary Medicine, Sudan University of Science and Technology, for routine ultrasonographic pregnancy diagnosis were examined. Ultrasound scanning was performed using a real-time scanner equipped with dual-frequency (3.5-5 MHz) curvilinear transducer.
RESULTS
The results showed that the prevalence of pseudopregnancy in goats in Khartoum State was 10.6%. Risk factors such as general body condition (=5.974; p=0.05), age (=11.760; p=0.0129), type of estrus (=12.794; p=0.000), and previous reproductive performance (=13.397; p=0.020) showed significant association (p≤0.05) with the occurrence of pseudopregnancy in the univariate analysis. Breed (=12.627; p=0.082), milk yield (=5.951; p=0.114), type of feeding (=1.721; p=0.190), season (=2.661; p=0.264), locality (=7.66; p=0.264), parity number (=0.451; p=0.767), and rearing system (=1.593; p=0.451) were not significantly associated with pseudopregnancy.
CONCLUSION
The prevalence of pseudopregnancy in goats in Khartoum State was 10.6%. Pseudopregnancy in goats is significantly associated with age, type of estrus, general body condition, and previous reproductive performance. This study showed for the first time that pseudopregnancy is a real reproductive problem in goats in Khartoum State.
PubMed: 29805220
DOI: 10.14202/vetworld.2018.525-529