Synthetic Strigolactone GR24 Improves Arabidopsis Somatic Embryogenesis through Changes in Auxin Responses

Somatic embryogenesis in Arabidopsis encompasses an induction phase requiring auxin as the inductive signal to promote cellular dedifferentiation and formation of the embryogenic tissue, and a developmental phase favoring the maturation of the embryos. Strigolactones (SLs) have been categorized as a novel group of plant hormones based on their ability to affect physiological phenomena in plants. The study analyzed the effects of synthetic strigolactone GR24, applied during the induction phase, on auxin response and formation of somatic embryos. The expression level of two SL biosynthetic genes, MOREAXILLARY GROWTH 3 and 4 (MAX3 and MAX4), which are responsible for the conversion of carotene to carotenal, increased during the induction phase of embryogenesis. Arabidopsis mutant studies indicated that the somatic embryo number was inhibited in max3 and max4 mutants, and this effect was reversed by applications of GR24, a synthetic strigolactone, and exacerbated by TIS108, a SL biosynthetic inhibitor. The transcriptional studies revealed that the regulation of GR24 and TIS108 on somatic embryogenesis correlated with changes in expression of AUXIN RESPONSIVE FACTORs 5, 8, 10, and 16, known to be required for the production of the embryogenic tissue, as well as the expression of WUSCHEL (WUS) and Somatic Embryogenesis Receptor-like Kinase 1 (SERK1), which are markers of cell dedifferentiation and embryogenic tissue formation. Collectively, this work demonstrated the novel role of SL in enhancing the embryogenic process in Arabidopsis and its requirement for inducing the expression of genes related to auxin signaling and production of embryogenic tissue.

The effect of Propofol versus Dexmedetomidine as anesthetic agents for oocyte pick-up on in vitro fertilization (IVF) outcomes

This study aimed to evaluate the effects of propofol and dexmedetomidine over different timescales on the IVF outcomes for transvaginal oocyte retrieval (TVOR). Twenty-four rats included in the study were divided into two main groups and three subgroups were subjected to the ovulation induction process. Group 1 was administered propofol (100 mg/kg i.v.) and group 2 were administered dexmedetomidine (25 µg/kg i.p.) The oviduct collection procedure was completed within 15 min for subgroup Pro15min, Dex15min (n = 4), within 16 to 30 min for subgroup Pro30min, Dex30min (n = 4) and within 31 to 60 min for subgroup Pro60min, Dex60min (n = 4) after euthanasia. The total number of oocytes was counted. After in vitro fertilization, the number and quality of embryos were evaluated. The number of pups born were evaluated after embryo transfer. The embryo number, quality and pup count decreased as the administration time for propofol increased (p < 0.05). No statistically significant difference was found between the dexmedetomidine subgroups for embryo number, quality and pup count(p > 0.05). As the exposure time to propofol increased, the number and quality of embryos obtained, and the pup count, decreased. The use of dexmedetomidine had no negative impacts on the number of embryos, their quality or the number of pups.

Effect of Female Age on Infertility Diagnostic Factors and In-vitro Fertilization Treatment Outcomes: A Single-center Retrospective Cohort Study

BackgroundInfertility has become an important issue in modern world because of the increasing number of infertile couples around the world. Advanced maternal age was considered to be a main factor of infertile problem. With second child policy published in China and more women in China intend to seek help for infertility problem, this study provided information of fertility diagnostic factors and IVF treatment outcomes of female IVF patients in different age groups, which can be a guidance for infertility diagnostic and treatment.MethodsA retrospective cohort study was conducted to IVF patient data collected by Jinjiang District Maternal and Child Health Hospital, Chengdu, China. Clinical and laboratory data of 45,743 fresh, autologous IVF cycles from January 2008 to September 2017 were included in the analysis. The diagnostic factors and treatment outcomes were analyzed for different age groups (age<35, n=30,708; age 35-41, n=11,921 and age≥42, n=3,114) as well as further stratified advanced age groups (age 42, n=933; age 43, n=744; age 44, n=556; age≥45, n=881). The characteristics including number of previous cycles, duration of infertility, BMI, basic FSH, basic AFC, AMH, retrieved oocyte number, fertilized oocyte number, transferred embryo number, baby number and economic cost were stratified according to patient age.ResultsThe basic characteristics of obesity rate, basic FSH and cancellation rate of IVF cycles in ≥42 years old group were significantly higher than other groups (p<0.01). Basic AFC, AMH, retrieved oocyte number, fertilized oocyte number and transferred embryo number in ≥42 years old group were significantly lower than other groups (p<0.01). Diagnostics characteristics and IVF-ET outcome declined significantly when maternal age increased (p<0.05). In the meanwhile, a preliminary analysis of cost per cycle showed that cost per child increased with patient’s age increase.ConclusionAlthough with increasing number of advanced maternal age IVF cycles, the age group of ≥42 years would intend to get unsatisfied outcome and higher cost per child. More guidance and considerations should be focused on encouraging earlier age treatment of infertility.Plain English summaryWith more and more women in the global range choose to get late pregnancy because of changes in society and economy, age has become an unavoidable factor in infertility diagnostic and treatment. Advanced age women may experience more infertility problems and negative IVF outcomes. A better understanding of the effect of maternal age on infertility would offer help to both diagnostic and treatment of IVF patient. This study conducted a single-center retrospective cohort analysis to female patients of different age groups and found that women with more advanced age (≥42) would be more easily to experience unsatisfied IVF outcome and higher economic cost to obtain one child. After the publication of second child policy in China in 2013, the number of advanced age patients increased, the necessity of special guidance for AMA patient may need to be taken into consideration.

Effect of Serum Progesterone (P4) Level at the Day of Human Chorionic Gonadotropin (HCG) Administration on the Outcome of Intracytoplasmic Sperm Injection and Fresh Embryo Transfer

Our aim was to investigate the effect of different progesterone (P4) at day of triggering HCG on the outcomes of ICSI with fresh embryo transfer in cases who underwent controlled ovarian stimulation (COS). Pituitary desensitization done either with long agonist or antagonist pituitary desensitization protocols, then analyzing the effect of serum P4-hCG level on ICSI-fresh embryos transfer outcome including; number and quality of retrieved oocytes (OR), embryo number (ER) & quality and ongoing pregnancy outcomes. The cases are which continued their pregnancy till 12 weeks, aborted cases, ectopic pregnancy & cancelled cases (for different reasons). Materials and Methods: This prospective cohort study was carried on 120 cases who underwent ICSI cycles for different causes and types of infertility. Controlled ovarian stimulation (COS) protocol and pituitary down-regulation either by; GnRH long agonist /antagonist protocols then, fertilization, embryo grading, embryo transfer and hormonal luteal support was done. Blood samples were taken on the day of hCG administration to measure P4 in all cases. All patients who got pregnant and continue till 12 weeks, they were categorized into 3 subgroups as regard serum P4 level: Group A: (P4< 0.5 ng/ml), Group B: (P4=O.5-I.5 ng/ml) & Group C: (P4 >I.5 ng/ml) and correlated with their ICSI outcome. The outcomes of ICSI-ET cycles in those cases were compared with 3 groups of P4 levels in the controlled ovarian stimulation with two protocols. Results: P4 level had insignificant relation with number and quality of retrieved oocytes, fertilized embryo number and quality and endometrial thickness regardless the protocol of COS. The low and high levels of P4 both, had a detrimental effect on CPR, meanwhile, with no harmful impact on the ongoing pregnancy rate was noticed, regardless the protocol of ovarian stimulation. P4-hCG isn’t considered to be the only predictive measure of ICSI outcome. Highest CRP was noticed in cases with P4 level (0.5-I.5 ng/ mI) regardless the type of protocol used.

Comparing the Individual and Combined Effects of Ant Attendance and Wing Formation on Aphid Body Size and Reproduction

Species employ multiple strategies to deal with stressful environments, but these strategies often incur costs. Aphids frequently utilize multiple predator avoidance strategies, including attracting mutualist ants for protection and dispersing by producing winged forms. While both strategies can be physiologically costly, the magnitudes of these costs have not been previously compared. In this study, we experimentally manipulated ant attendance in the field and measured the individual and interactive effects of ant attendance and wing formation on body size and reproduction of the ant-tended aphid Cinara schwarzii (Wilson) (Hemiptera: Aphididae). Aphid adult body size was smaller in the presence of ants (18%), but controlling for body size, there were no differences in embryo number or size. In contrast, wing formation did not affect adult body size but strongly reduced embryo number (46%) and size (8%). Although ant attendance reduced C. schwarzii wing formation, ant attendance and wing formation acted independently on aphid body size and reproduction. For comparison, we confirmed that the manipulation of ant presence had no effect on body size or reproduction of the untended co-existing congener Cinara solitaria (Gillette and Palmer) (Hemiptera: Aphididae). Complementing our empirical study, a meta-analysis of 78 responses from 24 publications showed that wing formation consistently and significantly reduces aphid body size and reproduction (37%), while the effects of ant attendance showed a mean positive effect (9%) that did not significantly differ from zero. Together, our empirical study and meta-analysis provide strong evidence for costs of wing formation but not ant attendance for aphids.

Practical application of an impractical bovine genotype: creating bilateral twin pregnancies in Trio allele carriers

Bovine twin birth is associated with detriments, including increased embryo/fetal losses, malpresentation, and dystocia. Incidence of these is lessened in bilateral compared with unilateral twin pregnancy. This study was undertaken to assess the use of follicular ablation by aspiration to create bilateral twin pregnancies in females with genetic potential for ~3.5 ovulations per cycle (Trio allele carriers). In experiment 1, carriers (n = 30) and noncarriers (n = 10) were synchronized for ovulation and timed artificial insemination (TAI). Follicles (&gt;5 mm) in excess of one per ovary were aspirated ~16 h preceding TAI. Follicle count for females with follicles on only one ovary was reduced to two. Blood was sampled 2 wk post-TAI to assess progesterone (P4) concentrations; embryo count was determined by ultrasound 6 wk post-TAI. Circulating P4 concentration post-TAI was significantly (P &lt; 0.001) associated with both genotype and subsequent pregnancy status (pregnant noncarriers: 7.06 ± 0.68 ng/mL; pregnant carriers: 5.54 ± 0.55 ng/mL; nonpregnant noncarriers: 5.22 ± 1.05 ng/mL; nonpregnant carriers: 3.13 ± 0.42 ng/mL). Experiment 2 was undertaken to offset the negative effects of follicular aspiration on subsequent P4 concentration observed in experiment 1. Carriers (n = 38) and noncarriers (n = 32) were submitted to TAI and follicle ablation as described for experiment 1. Additionally, accessory corpora lutea (CL) were induced in carriers by the administration of human chorionic gonadotropin (carriers) at day 6 post-TAI. Consequently, P4 concentration post-TAI was significantly (P &lt; 0.05) associated with subsequent pregnancy status (pregnant: 8.48 ± 0.61 ng/mL; nonpregnant: 6.70 ± 0.63 ng/mL) but not with genotype (carrier: 8.01 ± 0.59 ng/mL; noncarrier: 7.17 ± 0.64 ng/mL). Embryo number was greater in carriers (exp. 1: 1.64 ± 0.81; exp 2: 1.45 ± 0.09) vs. noncarriers (1.00 ± 0.00, both experiments). Single, twin, and triplet pregnancies occurred in carriers in experiment 1, whereas multiples in experiment 2 were limited to twin pregnancies. Genotype effects on pregnancy rate were not significant (P &gt; 0.10) in either experiment. Results suggest that follicular ablation to create bilateral twin pregnancies in Trio carriers is feasible but requires the induction of accessory CL to offset the negative effects of follicular aspiration on subsequent P4 concentration and associated fertility outcomes.

C57BL/6J and B6129F1 Embryo Transfer: Unilateral and Bilateral Transfer, Embryo Number and Recipient Female Background Control for the Optimization of Embryo Survival and Litter Size

Embryo transfer (ET) is a common procedure in rodent facilities. Optimizing this technique may help to reduce the number of animals, but little information is available regarding wild type strains and the conditions that affect embryo transfer. To explore this theme, 2-cell C57BL/6J embryos were transferred after overnight culture of freshly collected zygotes using different conditions: unilateral transfers using a total of 6, 8, 12, 15, 20 and 25 embryos were performed initially; then, this strain was also used for bilateral transfers using a total of 6, 12 and 20 embryos equally divided by the two oviducts. Groups of 25 embryos were not tested for the bilateral technique, since this condition produced the lower success rate when using the unilateral technique and 20 embryos would still represent a large number of embryos. A group of 2-cell B6129F1 embryos was also transferred using unilateral and bilateral ET with 6, 12 and 20 embryos. Crl:CD1(ICR) were used as recipient females for non-reciprocal transfers and C57BL/6J were used to test reciprocal transfers (only tested for six C57BL/6J unilateral transfers). Unilateral transfers using C57BL/6J mice produced higher success rates using six embryos, compared to the other groups transferred unilaterally (p-values between 0.0001 and 0.0267), but the mean number of pups per litter was not different among groups. Bilateral transfer produced higher number of pups when 20 embryos were divided by the two oviducts compared to six (p = 0.0012) or 12 (p = 0.0148) embryos, but with no differences in success rates. No statistical differences were found between the groups of B6129F1, but better results were obtained on bilateral transfers using a total of six embryos. For the strain tested (C57BL/6J), the uterine environment (Crl:CD1(ICR) or C57BL/6J recipient) does not impact the outcome of the technique. These results complement previous work published using genetically engineered mice strains and show that unilateral transfers using low number of embryos (6), produce better outcomes when compared to bilateral or unilateral transfers using more embryos. It also highlights differences between the outcome of bilateral transfers in the two strains tested. A set of historical data of genetically engineered mice at a C57BL/6J background was also included, confirming that lower embryo numbers are related to higher success rates. Together, the outcome of these experiments can be important to reduce the number of recipient and donor females, optimize embryo transfers and improve animal welfare discouraging the use of a more invasive technique.

The therapeutic effect of interleukin-22 in high androgen-induced polycystic ovary syndrome

Polycystic ovary syndrome (PCOS) is a complex syndrome involving both endocrine and metabolic disorders. Gut microbiota and the intestinal immune factor IL-22 play an important role in the pathogenesis of PCOS. However, the therapeutic role of IL-22 in high androgen-induced PCOS mice is not clear. We aimed to determine the therapeutic effects of IL-22 on the DHEA-induced PCOS mouse model and to explore the possible mechanism of IL-22 in regulating hyperandrogenism-associated PCOS. Insulin resistance levels and ovarian functions were investigated in DHEA-induced PCOS mice with or without additional IL-22 treatment. We found that IL-22 could reverse insulin resistance, disturbed estrous cycle, abnormal ovary morphology, and decreased embryo number in DHEA mice. Mechanistically, IL-22 upregulated the browning of white adipose tissue in DHEA mice. This study demonstrated that IL-22-associated browning of white adipose tissue regulated insulin sensitivity and ovarian functions in PCOS, suggesting that IL-22 may be of value for the treatment of PCOS with a hyperandrogenism phenotype.

Reproductive traits of deep-sea armoured shrimp, Glyphocrangon investigatoris from Bay of Bengal, Indian Ocean

Details on size at first maturity, embryo number and size, brood chamber volume and reproductive output of deep-sea armoured shrimp, Glyphocrangon investigatoris caught off the south-east coast of India by using EXPO trawl from 633 m depth in FORV ‘Sagar Sampada’ are reported here. Eighty-four female shrimps ranging from 17.29–36.31 mm carapace length and 2.28–16.54 g weight formed 7.73% of total catch, 30% of which was constituted by embryo-bearing females. Regression of weight on carapace length revealed negatively allometric growth (r2 = 0.85, P < 0.01). The size at first maturity was estimated as 19.96 mm. The embryo number ranged from 55 to 233 with a mean of 120.24 ± 34 embryos and showed a positive correlation to body size. Embryo diameter varied between 1.0 to 3.34 mm and more than 50% of embryos constituted the 2.0–2.5 mm size class. Brood chamber volume and percentage frequency of embryo stage development revealed a linear relationship with carapace length. Based on dry weight, mean reproductive output was estimated to be 0.16. The female armoured shrimps showed a high reproductive investment evidenced from few, large, yolky embryos, indicating their deep-sea adaptation.

Teratogenic Effect of Congenital Toxoplasmosis in Chicken Embryo

This research is designed to observe the teratogenic effect of Toxoplasma gondii infection in chick embryos, based on the number of somites, embryo length and the development of embryonic brain vesicles. Methods in the research: Chicken eggs were infected with 1 x 103 tachyzoites of T. gondii. The eggs were incubated in eggs hatching box. Observation of somite performed on embryonated eggs 24 hours after incubation and the embryonic development of vesicles performed 72 hours after incubation then the length of each embryo were measured. Results: Revealed that there was a significant difference in the number of somites (p < 0.1), T. gondii infection reduced the number of somites. While in the number of brain vesicles in 3 - days old chicken embryos, although there was no significant difference, the size declining emerged. The length of the embryos both at 24 or 72 hours old showed that T. gondii infection reduced the length (p < 0.1). Conclusions: T. gondii infection influences the development of chicken embryos in the declining of length and the decreasing of somite embryo number. Keywords: IGF-I crossbreed mare serum pregnant; Follicle; Mus musculus