Days Of Pregnancy Research Articles

D-galactose causes embryonic development arrest and placental development disorders in mice by increasing ROS and inhibiting SIRT1/FOXO3a axis

IntroductionDoes an elevation in d-Galactose (D-Gal) levels within the body contribute to abnormal embryonic development and placental dysfunction during pregnancy? MethodsMouse embryos were cultivated to the blastocyst stage under varying concentrations of D-Gal. The blastocyst formation rate was measured, and the levels of reactive oxygen species (ROS), sirtuin 1 (SIRT1), and forkhead box O3a (FOXO3a) in blastocysts were assessed. Mice were intraperitoneally injected with either saline or D-Gal with or without SRT1720. On the 14th day of pregnancy, the fetal absorption rate and placental weight were recorded. Placental levels of superoxide dismutase (SOD) and malondialdehyde (MDA) were determined. The expression of senescence-related factors, such as senescence-associated β-galactosidase (SA-β-gal) in the placenta was examined, and the expression of placental SIRT1, FOXO3a and p21 was evaluated by immunohistochemistry and Western blotting. ResultsD-Gal adversely affects early embryonic development in vitro, resulting in a decreased blastocyst formation rate. Furthermore, D-Gal downregulates SIRT1 and FOXO3a while increasing ROS levels in blastocysts. Concurrently, D-Gal induces placental dysfunction, characterized by an elevated fetal absorption rate, reduced placental weight, diminished SOD activity, and increased MDA content. The senescence-related factor SA-β-gal was detected in the placenta, along with altered expression of placental SIRT1, FOXO3a, and p21. The SIRT1 agonist SRT1720 mitigated this damage by increasing SIRT1 and FOXO3a expression. DiscussionThe inhibition of early embryonic development and placental dysfunction induced by D-Gal may be attributed to the dysregulation of SIRT1. Activating SIRT1 emerges as a potentially effective strategy for alleviating the adverse effects of D-Gal exposure.

Neurodevelopmental effects of prenatal Bisphenol A exposure on the role of microRNA regulating NMDA receptor subunits in the male rat hippocampus

Maternal bisphenol A (BPA) exposure has been reported to cause learning and memory deficits in born offspring. However, little is known that this impairment is potentially caused by epigenetic modulation on the development of NMDA receptor subunits. This study investigates the effect of prenatal BPA exposure on the hippocampal miR-19a and miR-539, which are responsible for regulating NMDA receptor subunits as well as learning and memory functions. Pregnant Sprague Dawley rats were orally administered with 5 mg/kg/day of BPA from pregnancy day 1 (PD1) until gestation day 21 (GD21), while control mothers received no BPA. The mothers were observed daily until GD21 for either a cesarean section or spontaneous delivery. The male offspring were sacrificed when reaching GD21 (fetus), postnatal days 7, 14, 21 (PND7, 14, 21) and adolescent age 35 (AD35) where their hippocampi were dissected from the brain. The expression of targeted miR-19a, miR-539, GRIN2A, and GRIN2B were determined by qRT-PCR while the level of GluN2A and GluN2B were estimated by western blot. At AD35, the rats were assessed with neurobehavioral tests to evaluate their learning and memory function. The findings showed that prenatal BPA exposure at 5 mg/kg/day significantly reduces the expression of miR-19a, miR-539, GRIN2A, and GRIN2B genes in the male rat hippocampus at all ages. The level of GluN2A and GluN2B proteins is also significantly reduced when reaching adolescent age. Consequently, the rats showed spatial and fear memory impairments when reaching AD35. In conclusion, prenatal BPA exposure disrupts the role of miR-19a and miR-539 in regulating the NMDA receptor subunit in the hippocampus which may be one of the causes of memory and learning impairment in adolescent rats.

Effect of Shoutai Pills on expression of key glycolytic proteins and apoptosis related factors at maternal fetal interface in mouse model of threatened abortion with syndrome of kidney deficiency

This study aims to explore the mechanism of Shoutai Pills in treating threatened abortion. According to the random number table method, ICR female mice were randomized into a normal group, a model group, a dydrogesterone group, and a Shoutai Pills group, with 15 mice in each group. Mice were administrated with normal saline(normal and model groups) or the suspension of Shoutai Pills or dydrogesterone by gavage at 9:00 am every day. At 16:00 every day, mice in the normal group were administrated with an equal volume of distilled water, while those in the model, Shoutai Pills, and dydrogesterone groups were administrated with hydrocortisone solution by gavage for 4 consecutive days. ICR female and male mice were caged in a ratio of 2∶1 during the pre-estrous or estrous period. From the first day of pregnancy, drug administration was continued for 5 consecutive days. On day 6, mice were administrated with mifepristone by gavage to establish the model of kidney deficiency-induced abortion. On day 6 of pregnancy, 10 female ICR mice were randomly selected from each group, and the uterus was collected for observation of the pathological changes of trophoblasts at the maternal-fetal interface by hematoxylin-eosin(HE) staining. The protein levels of key enzymes of glycolysis, hexokinase 2(HK2), enolase 1(ENO1), pyruvate kinase M2(PKM2), and lactate dehydrogenase A(LDHA), were determined by Western blot and immunofluorescence. The expression of apoptosis-related proteins including B cell lymphoma-2(Bcl-2), Bcl-2-associated protein X(Bax), and cysteinyl aspartate-specific proteinase-3(caspase-3) was determined by Western blot and real-time PCR. Terminal-deoxynucleoitidyl transferase-mediated nick-end labeling was employed to examine apoptosis. The embryo loss rate of the remaining five female mice was calculated by trypan blue staining method on day 14 of pregnancy. On day 14 of pregnancy, the embryo loss rate of the normal group was 5.00%, which was lower than that(27.78%) in the model group(P<0.05). Dydrogesterone and Shoutai Pills groups showed reduced embryo loss rates(10.26% and 7.50%, respectively) compared with the model group. On day 6 of pregnancy, compared with the normal group, the model group showed down-regulated expression of HK2, ENO1, PKM2, LDHA, and Bcl-2 and up-regulated expression of Bax and caspase-3(P<0.05). Compared with the model group, dydrogesterone and Shoutai Pills up-regulated the expression of HK2, ENO1, PKM2, LDHA, and Bcl-2 and down-regulated the expression of Bax and caspase-3(P<0.05). Compared with that in the normal group, the apoptosis rate in the model group increased(P<0.05). Compared with the model group, dydrogesterone and Shoutai Pills reduced the apoptosis rate(P<0.05). In conclusion, Shoutai Pills can reduce the embryo loss rate and protect embryos by promoting aerobic glycolysis at the maternal-fetal interface and inhibiting the apoptosis of trophoblasts in mice.

The Teratogenic Effect of Dimefluthrin-Based Mosquito Coils on Pregnant Mice (Mus musculus L.)

Dimefluthrin, a synthetic pyrethroid insecticide, claims to be safe and harmless to mammals. However, the potential risk of mosquito coils containing dimefluthrin to pregnant mice remains uncertain. This study aimed to examine the teratogenic effect of dimefluthrin when inhaled by pregnant mice at different stages of gestation. The study used mosquito coils with 0.031% dimefluthrin as the active ingredient and was exposed to pregnant mice at various pregnancy stages. The results revealed that mosquito coils with dimefluthrin had teratogenic effects, reducing the number of live fetuses and increasing the number of dead fetuses, especially from the seventh to the eighteenth day of pregnancy. Additionally, there was a decrease in fetal weight, length, and volume, particularly during the seventh to eighth day of gestation. Hemorrhages were observed as fetal abnormalities and were highest three hr after exposure every day during pregnancy. The conclusion of this research is pregnant mice exposed to mosquito coil smoke containing dimefluthrin experienced a decrease in the number of live fetuses and an increase in fetal mortality. While there was no significant impact on reabsorption, the fetuses showed reduced weight, and size, and an increased incidence of hemorrhages when exposed for three hr daily during pregnancy.

Immunohistochemical Investigation of Autophagy in the Uterus during the First Trimester of Pregnancy in Rats

During placental development, autophagy has an important role at the molecular level, especially in cases such as trophoblast cell proliferation and cell death. Abnormal placental development due to trophoblast dysfunction causes serious gynaecological diseases and various fetal malformations. In the study conducted to investigate autophagy on the 5th day of pregnancy, in the pregnant and non pregnant group uterus tissues, uterine glands LC3 and Beclin 1 (+), in the evaluation of myometrium and perimetrium, weak (+) was observed in myometrium cells in pregnancy, while (+) reaction could not be distinguished in perimetrium. In this study, it was concluded that the immunohistochemical increase in LC3 and Beclin 1 intensity in the uterus, especially in the endometrial areas in the first trimester of pregnancy compared to the control group tissues is related to the physiology of pregnancy, homeostasis in pregnancy and hormonal mechanism.

Fetal protective effect of Indonesian propolis from Apis mellifera against rifampicin-pyrazinamide induced impaired pregnancy in BALB/c mice.

Anti-tuberculosis drugs rifampicin and pyrazinamide combination in pregnancy can cause morphological, visceral and skeletal damage. Several studies showed that propolis improves pregnancy outcomes. This study aims to determine the fetal protective effect of propolis in BALB/c mice given the anti-tuberculosis drug combination rifampicin and pyrazinamide. A total of 21 pregnant mice were randomly divided into three groups: the normal group (N)was given distilled water as a vehicle, the positive control group (RP) were given rifampicin 15 mg/kg BW, pyrazinamide 35 mg/kg BW and the treatment group (IP) were given rifampicin 15 mg/kg BB, pyrazinamide 35 mg/kg BW and propolis 400 mg/kg BW. The treatment was given during the period of organogenesis, from day 6 to day 15. Laparotomy was performed on the 18th day of pregnancy. Maternal and fetal body weight, fetal length, number of fetuses, and skeletal defects of fetuses were used as parameters to identify the teratogenic effect. All data were analyzed using the ANOVA. All groups significantly differed between maternal and fetal body weights (p<0.05). The administration of rifampicin-pyrazinamide and propolis during pregnancy did not significantly affect the number of fetuses (p>0.05). The administration of propolis protects the fetus from skeletal abnormalities. While in the RP and IP groups, we can find resorption sites and haemorrhagic. This study may suggest the protective effects of propolis against rifampicin pyrazinamide-induced impaired pregnancy.

Inactivated Flagellin-Containing Vaccine Efficacy against Ovine Enzootic Abortion.

Chlamydia abortus is the etiological agent of abortion and fetal loss in sheep, goats and bovine cattle in many countries. Even though commercially available vaccines can reduce the incidence in sheep, the development of new, safe, and effective vaccines remains high on the agenda. In this study, an evaluation was made of the efficacy of a vaccine candidate, an inactivated antigen based on the extract of outer membrane proteins of a C. abortus strain known as Chlamydia VNITIBP-21, in combination with recombinant flagellin as an adjuvant. Pregnant sheep (n = 43) were divided into three groups: an experimental vaccinated group, a control infected group and a control non-infected group. The sheep were vaccinated twice, with an interval of 3 weeks, then infected with the homologous virulent strain of Chlamydia abortus on pregnancy day 75. The vaccine candidate reduced C. abortus shedding in vaginal swabs considerably, in comparison with the control group. In addition, ewes in the experimental group experienced no abortions, while those in the control group experienced instances of abortion, as well as births of weak and nonviable lambs. The findings show that the vaccine candidate proved itself to be promising in combatting the agent of ovine abortion and fetal loss.

Ultrasound markers of adverse pregnancy outcomes in embryonal period: literature review and own clinical cases

A goal of the review was to search and describe all currently known prenatal ultrasound markers of adverse pregnancy outcomes. The review is instantiated by multiple ultrasound images of own clinical cases. Taking into account the authors wide experience, it is possible to use all of presented ultrasound signs as predictors (markers) of an adverse pregnancy outcome in the early stages. From our point of view, it is reasonable to divide the markers into two groups, “primary markers” and “dynamic markers”. In the case of “Primary markers” revealing on initial ultrasound, it is reasonable to perform control ultrasound in 7–10 days to confirm the viable pregnancy or missed miscarriage. The “Dynamic markers” can be used on control ultrasound, and some of them may be used in shorter dynamic period when there is a need to predict the outcome earlier than 7-10 days. Generally, the review describes 22 early pregnancy ultrasound markers of adverse pregnancy outcome. These are imaging features of the gestational sac, yolk sac, embryo size, embryo heartbeat, and amnion. Early pregnancy ultrasound is not mandatory, but in a real-case scenario, almost all women undergoes ultrasound on early stage of pregnancy in objective or subjective causes. Therefore, in the first days of pregnancy, women should receive a highly professional diagnostic by a physician, able to assess the risks and prognosis of pregnancy outcome. The authors of the review particularly notes that the assessment of ultrasound markers of adverse pregnancy outcomes must be carried out with the strictest adherence to the medical ethics, deontology, and medical privacy.

Adaptive changes in immunological, hematological and metabolic profiles of pregnant cows

Pregnancy triggers a series of complex interactions within the body, involving both the immune system and metabolic processes. In the case of pregnant cows, these mechanisms are essential for supporting the developing fetus and maintaining the overall health of the mother. The aim of the work was to study the hematological, biochemical, and immunological parameters of cows at different periods of pregnancy. Research was conducted on cows (Bos Taurus) of the Ukrainian Black-and-White dairy breed. Groups were formed based on the principle of analogs. The research was carried out using the methodology of group-period experiments. The control group consisted of non-pregnant cows. The first observation period was conducted on pregnant cows with confirmed pregnancy (30 days). The second observation period was carried out at 90 days of pregnancy, the third at 180 days of gestation, and the fourth at 270 days. Laboratory studies included immunological research conducted in the specialized laboratory of immunology for reproduction animals. The first notable observation was the gradual increase in the population of CD3+ lymphocytes from the beginning of gestation, which remained stable until the 90th day of pregnancy and then gradually decreased. The population of B cell immune cells CD22+ had its peculiarities: dynamic and stable growth from the start of pregnancy until the middle of the second trimester (180 days), followed by a gradual decrease until the onset of labor. The redistribution of T and B lymphocytes occurred with dynamic fluctuations in the number of undifferentiated 0-lymphocytes. The processes of transformation and adaptation of homeostasis affected both enzyme activity and indicators of mineral metabolism. Thus, from the beginning of pregnancy (from the 30th day of gestation), the experimental cows showed a significant increase in alkaline phosphatase activity (P &lt; 0.05), alanine aminotransferase (P &lt; 0.05), and aspartate aminotransferase activity (P &lt; 0.05). During the pregnancy of cows, body changes occur. From the early period of gestation throughout the entire period of embryogenesis, changes in the system of hematopoiesis, alterations in the activity of enzymes, exchange of Ca and P, accompanied by changes in immunological reactions take place. These processes are adaptive and ensure the maintenance of the homeostasis system. The perspective of further research will aim at studying changes in immunological and biochemical parameters during physiological pregnancy and when it is complicated.

Maternal probiotic Lactocaseibacillus rhamnosus HN001 treatment alters postpartum anxiety, cortical monoamines, and the gut microbiome

Peripartum mood and anxiety disorders (PMADs) affect 15–20% of peripartum women and are well known to disrupt infant caregiving. A recent study in humans reported that anxiety and depressive symptoms were alleviated by peripartum treatment with the probiotic, Lactocaseibacillus rhamnosus HN001. The current study determined the effects of chronic Lactocaseibacillus rhamnosus HN001 (HN001) treatment on postpartum affective and caregiving behaviors in a laboratory rodent model. Female rats were given probiotic overnight in their drinking water, or untreated water, from the first day of pregnancy through postpartum day 10. To determine whether the HN001 effects were influenced by a background of stress, half the females underwent chronic variable pregnancy stress and the other half remained undisturbed. The results revealed that, even without pregnancy stress, HN001 reduced postpartum anxiety-related behavior, increased variability in behavioral fragmentation when dams interacted with pups, increased time away from pups, and decreased prefrontal cortex norepinephrine (NE), dopamine (DA) and serotonin (5-HT). Probiotic plus stress consistently reduced the latency to float in the forced swim test, increased DA and 5-HT turnovers in the prefrontal cortex, increased hippocampal NE, and reduced hypothalamic DA. Fecal microbe alpha and beta diversities were lower postpartum than prepartum, which was prevented by the probiotic treatment and/or stress. Across the entire sample lower postpartum anxiety behavior was associated with lower fecal Bacteroides dorei. This study reveals novel information about how L. rhamnosus HN001 influences postpartum behavior and microbiota-gut-brain physiology in female laboratory rats, with implications for probiotic supplement use by pregnant and postpartum women.

Effect of pre and postpartum maternal supplementation of zinc and selenium via slow-release glass bolus or the element salts on feed intake and some blood parameters in ewes and their lambs

In late pregnancy and early lactation, ewe's demand increases for zinc (Zn), and selenium (Se) for scavenging of free radicals and due to transfer via placenta, colostrum or milk to lamb.This experiment was conducted to study the effect of slow-release bolus of Zn and Se on performance, and some blood parameters of pregnant ewes in the last third of pregnancy, and early lactation as well as their lambs. Twenty-one ewes were estrus synchronized in the breeding season by intra vaginal insertion of sponge containing medroxyprogesterone acetate. Forty-five days before the expected date of parturition, the pregnant ewes (3–4 years old and 55±5.2 kg in weight) were randomly assigned to one of the three groups: 1) control, 2) slow-release glass bolus with mean release of 20 mg Zn and 0.2 mg Se per day (Bolus), and 3) 20 mg Zn and 0.2 mg Se per day as sulfate and selenite, respectively via Daily feeding (Daily). Blood samples were obtained from ewes at 105th and 135th days of pregnancy, at parturition and 15th day postpartum and from lambs at 15 days of age. The results indicated that total feed intake of ewes increased (P = 0.03) in the Bolus and Daily groups than control. Supplementation of Zn and Se in both methodssignificantly increased Total Antioxidant Capacity (TAC) in the ewesand their lambs than the control group. Compared with the control, there was higher triiodothyronine (T3) in ewes and their lambs and lower tetraiodothyronine (T4) only in lambs in Bolus and Dailygroups. Increased T3 to T4 ratio in ewes (at days 135 of pregnancy and at parturition) and their lambs was recorded in Bolus and Daily than the control group. An increased level of alkalinephosphatase (ALP) in ewes was observed in the Bolus and Daily groups compared with the control. Alanine amino transferase (ALT) activity on day 135 of pregnancy was significantly reduced by Zn and Se supplementation in both methods. Bolus and Daily groups of ewes had lower creatinephosphokinase (CPK), and their lambs had lower aspartate amino transferase (AST), ALT, CPK, and gamma-glutamyltransferase (GGT) than the control group. Regardless of the supplementation method, Zn and Se increased glucose (P = 0.02) and decreased triglyceride (P = 0.06) in ewes. In the ewes supplemented with Zn and Se, creatinine concentration significantly decreased in parturition day compared to control. Total protein, albumin, globulin and ratio of albumin to globulin in ewes and their lambs were not different between treatments. As a result, Zn and Se improved antioxidant status and some blood parameters in ewes and their lambs. Efficacy of slow-release bolus of Zn and Se was similar with daily feeding of these elements.

The neuroprotective effect of exogen melatonin upon fetal hippocampus damage caused by high-dose caffeine administration in pregnant rats.

The aim of this study is to evaluate whether exogenous melatonin (MEL) mitigates the deleterious effects of high-dose caffeine (CAF) administration in pregnant rats upon the fetal hippocampus. A total of 32 adult Wistar albino female rats were divided into four groups after conception (n = 8). At 9-20 days of pregnancy, intraperitoneal (i.p.) MEL was administered at a dose of 10mg/kg/day in the MEL group, while i.p. CAF was administered at a dose of 60 mg/kg/day in the CAF group. In the CAF plus MEL group, i.p. CAF and MEL were administered at a dose of 60 and 10mg/kg/day, respectively, at the same period. Following extraction of the brains of the fetuses sacrificed on the 21st day of pregnancy, their hippocampal regions were analyzed by hematoxylin and eosin and Cresyl Echt Violet, anti-GFAP, and antisynaptophysin staining methods. While there was a decrease in fetal and brain weights in the CAF group, it was found that the CAF plus MEL group had a closer weight average to that of the control group. Histologically, it was observed that the pyramidal cell layer consisted of 8-10 layers of cells due to the delay in migration in hippocampal neurons in the CAF group, while the MEL group showed similar characteristics with the control group. It was found that these findings decreased in the CAF plus MEL group. It is concluded that high-dose CAF administration causes a delay in neurogenesis of the fetal hippocampus, and exogenous MEL is able to mitigate its deleterious effects.

Use of leflunomide among girls and women of childbearing age and occurrence of leflunomide-exposed pregnancies in Germany

Leflunomide is contraindicated during pregnancy and treatment cessation is recommended two years before pregnancy. We aimed to describe leflunomide use in women of childbearing age in Germany, the occurrence of pregnancies in women using leflunomide and malformations among children possibly exposed in utero. Using the GePaRD database (claims data, ∼20% of the German population), we determined annual age-standardized prevalences of leflunomide use between 2004 and 2019 among females aged 13–49 years. Further, we estimated the number of exposed pregnancies by assessing whether the exposure window assigned to the last dispensation before pregnancy (days covered by the dispensation plus two years) overlapped the onset of pregnancy or whether there was a dispensation in the first eight weeks of pregnancy. For exposed live births, a mother-baby linkage was performed and the presence of congenital malformation was assessed. The age-standardized prevalence of leflunomide use ranged between 0.34 and 0.46 per 1000 females during the study period. About one third of the users were ≤40 years. We identified 205 leflunomide-exposed pregnancies ending during the study period. 71% of these pregnancies ended in a live birth (26% preterm) and 10% in an induced abortion. In 86% of the live births (n=125) the mother-baby linkage was successful. Among these 125 children, 13 children (10%) had congenital malformations. In conclusion, we observed a considerable number of pregnancies in women using leflunomide in the two years before or during early pregnancy. This highlights the importance of monitoring the implementation of existing risk minimization measures for leflunomide in Germany.

18 Progesterone and insulin concentrations during pregnancy in slick and wild type-haired Holstein cows

Abstract In cattle, the presence of the SLICK gene has been associated with shorter calving intervals when exposed to tropical climates. However, the effects this genotype may have in terms of circulating hormones during pregnancy have not been reported. Therefore, the current study aimed to determine if reproductive hormones concentrations differ during the gestation between slick (SH) and wild type-haired (WT) cattle. Holstein heifers were estrus synchronized and inseminated during Winter (December) and Summer (June). A total of 13 SH and 8 WT Holstein heifers became pregnant and are included in the current study. Beginning on the day of insemination (d 0), blood samples were collected every 28 d. Commercially available ELISA kits were used to determine the plasma concentrations of bovine progesterone and insulin. The effects of hair coat genotype, season, day of pregnancy, and the respective interactions were tested by PROC Mixed in SAS. Day of pregnancy was utilized as a repeated measure for each animal. A significant season x day of pregnancy interaction (P &amp;lt; 0.001) was observed for the plasma concentrations of progesterone. On gestational d 0, 28 and 56, progesterone concentrations were greater for heifers inseminated during the Winter (7.11 ± 0.50, 5.31 ± 0.51, 5.39 ± 0.49 ng/mL, for d 0, 28 and 56 of pregnancy, respectively) when compared with those inseminated during the Summer (3.04 ± 0.45, 3.17 ± 0.45, 3.21 ± 0.45 ng/mL, for d 0, 28 and 56 of pregnancy, respectively). For insulin, also a significant interaction of season x day of pregnancy was observed (P = 0.004). On d 112 of pregnancy, the concentrations of insulin were greater for heifers inseminated during Winter (0.337 ± 0.03 ng/mL) when compared with those inseminated during Summer (0.082 ± 0.08 ng/mL). No other significant interactions were observed. Neither differences in progesterone (P = 0.70; 3.7 ± 0.33 and 3.9 ± 0.25 ng/mL, for WT and SH, respectively) nor insulin (P = 0.46; 0.20 ± 0.02 and 0.18 ± 0.01 ng/mL, for WT and SH, respectively) were observed between genotypes. Further research comparing other hormones during pregnancy may provide useful information between genotypes that may help to explain the better reproductive performance of SH dairy cows.

Reference Gene Validation in the Embryonic and Postnatal Brain in the Rat Hyperhomocysteinemia Model.

Maternal hyperhomocysteinemia (HCY) induced by genetic defects in methionine cycle enzymes or vitamin imbalance is known to be a pathologic factor that can impair embryonal brain development and cause long-term consequences in the postnatal brain development as well as changes in the expression of neuronal genes. Studies of the gene expression on this model requires the selection of optimal housekeeping genes. This work aimed to analyze the expression stability of housekeeping genes in offspring brain. Pregnant female Wistar rats were treated daily with a 0.15% L-methionine solution in the period starting on the 4th day of pregnancy until delivery, to cause the increase in the homocysteine level in fetus blood and brain. Housekeeping gene expression was assessed by RT-qPCR on whole embryonic brain and selected rat brain areas at P20 and P90. The amplification curves were analyzed, and raw means Cq data were imported to the RefFinder online tool to assess the reference genes stability. Most of the analyzed genes showed high stability of mRNA expression in the fetal brain at both periods of analysis (E14 and E20). However, the most stably expressed genes at different age points differed. Actb, Ppia, Rpl13a are the most stably expressed on E14, Ywhaz, Pgk1, Hprt1 - on E20 and P20, Hprt1, Actb, and Pgk1 - on P90. Gapdh gene used as a reference in various studies demonstrates high stability only in the hippocampus and cannot be recommended as the optimal reference gene on HCY model. Hprt1 and Pgk1 genes were found to be the most stably expressed in the brain of rat subjected to HCY. These two genes showed high stability in the brain on E20 and in various areas of the brain on the P20 and P90. On E14, the preferred genes for normalization are Actb, Ppia, Rpl13a.

Comparative effectiveness of different methods of using the analogue of gonadotropin-releasing hormone for ovulation stimulation in rabbit does

Modern reproductive technologies require artificial insemination (AI) protocols that are safe for the health of animals and do not violate their welfare. Stress factors arising in the reproduction process of industrial rabbit breeding negatively affect the sexual function and reproductive performance of rabbits and their offspring. In recent years, researchers have focused on alternative methods of ovulation stimulation, including intravaginal introduction of hormonal means of its induction. Therefore, the aim of our study was a comparative evaluation of the effectiveness of the intramuscular method of administration of the gonadotropin-releasing control (GnRH) analogue and its addition to the sperm dose for AI on Hyplus rabbit (n = 30), which were randomly divided into three groups. For this, buserelin acetate was administered intramuscularly at a dose of 1 μg per rabbit immediately after artificial insemination (control group). Experimental animals were inseminated with sperm doses with the addition of a GnRH analogue in the amount of 25 μg (experimental group 1) and 15 μg (experimental group 2). The influence of the method of administration of GnRH analogue on reproductive function was evaluated by indicators of the effectiveness of AI, fertility, and the dynamics of the progesterone level during pregnancy. A decrease in fecundity was observed in females; for example, in experimental group 1, the effectiveness of AI in terms of the number of rabbits was 64.3 %; in animals of experimental group 2 – 41.2 %, and the number of inseminations in this group of rabbits was 41.7% higher compared to control data. The fertility of rabbits of all groups was high, reaching the maximum value in the control group (10.42 ± 0.52 heads/female). Fertility according to live newborns in experimental group 1 was the highest among all groups of rabbits and was 9.67 ± 0.49 heads/female, which showed an increasing trend compared to the control. Also, in the animals of experimental group 1, an increase in the level of progesterone in blood serum was observed (on the 7th day of pregnancy by 13.2 %, on the 14th day by 14.3 %, on the 21st day by 16.4 %). In contrast, in experimental group 2, a significant decrease in the progesterone level was noted. Thus, 25 μg of GnRH analogue added to the extender is optimal. It is worth noting that there were no advantages of intravaginal administration over intramuscular administration in terms of reproductive performance. However, the number of live births and progesterone levels were improved.

Exercise and insulin glargine administration in mothers with diabetes during pregnancy ameliorate function of testis in offspring: Consequences on apelin-13 and its receptor

AimsDespite the evidence exhibited that diabetes during gestation (DDG) is linked with reproductive dysfunction in offspring, the underlying cellular mechanisms involved are not precisely defined. This study was designed to assess the impact of voluntary exercise and insulin glargine on DDG-induced metabolic and reproductive disorders in male offspring. Main methodsFifty female Wistar rats (three weeks old) received a control diet (n = 10) or high-fat-high-sucrose diet (to induce DDG; n = 40) for six weeks before breeding. From the 7th day of pregnancy onwards, blood glucose over 140 mg/dL was characterized as DDG. Then, the DDG animals were randomly divided into four subgroups with/without voluntary exercise and/or insulin glargine. To evaluate insulin resistance, a glucose tolerance test was performed on the 15th day of pregnancy. After three weeks, male offspring were weaned, and fed a control diet until 12 weeks old. At the end of the experiment, the lipid profile, sex hormones, and apelin-13 in the serum, mRNA expression of apelin receptors (APJ) in the testis and sperm analysis were assessed. Key findingsOur results indicated that voluntary exercise and/or insulin glargine administration in mothers with DDG ameliorated lipid profile, and sex hormones alterations, reduced the serum level of apelin-13, as well as increased APJ expression in testis, and quality of sperm in offspring. SignificanceCombined administration of voluntary exercise and insulin glargine during pregnancy by regulating of apelinergic system and inhibiting the metabolic and reproductive complications induced by DDG, can be considered as a suitable therapeutic strategy for improving sub-or in-fertility in the male offspring.

Maternal exposure to polystyrene nanoplastics causes defective retinal development and function in progeny mice by disturbing metabolic profiles

Microplastics (MPs) and nanoplastics (NPs) are widely spreading in our living environment, accumulating in the human body and potentially threating human health. The retina, which is a terminally differentiated extension of the central nervous system, is essential for the visual system. However, the effects and molecular mechanisms of MPs/NPs on retina development and function are still unclear. Here, we investigated the effects and modes of action of polystyrene NPs (PS-NPs) on the retina using mice as a mammalian model species. Maternal PS-NP exposure (100 nm) at an environmentally realistic concentration of 10 mg L−1 (or 2.07 *1010 particles mL−1) via drinking water from the first day of pregnancy till the end of lactation (21 days after birth) caused defective neural retinal development in the neonatal mice, by depositing in the retinal tissue and reducing the number of retinal ganglion cells and bipolar cells. Exposure to PS-NPs retarded retinal vascular development, while abnormal electroretinogram (ERG) responses and an increased level of oxidative stress were also observed in the retina of the progeny mice after maternal PS-NP exposure. Metabolomics showed significant dysregulation of amino acids that are pivotal to neuron retinal function, such as glutamate, aspartate, alanine, glycine, serine, threonine, taurine, and serotonin. Transcriptomics identified significantly dysregulated genes, which were enriched in processes of angiogenesis, visual system development and lens development. Regulatory analysis showed that Fos gene mediated pathways could be a potential key target for PS-NP exposure in retinal development and function. Our study revealed that maternal exposure to PS-NPs generated detrimental effects on retinal development and function in progeny mice, offering new insights into the visual toxicity of PS-NPs.

Bioinformatic analysis of endometrial miRNA expression profile at day 26–28 of pregnancy in the mare

The establishment of the fetomaternal interface depends on precisely regulated communication between the conceptus and the uterine environment. Recent evidence suggests that microRNAs (miRNAs) may play an important role in embryo-maternal dialogue. This study aimed to determine the expression profile of endometrial miRNAs during days 26–28 of equine pregnancy. Additionally, the study aimed to predict target genes for differentially expressed miRNAs (DEmiRs) and their potential role in embryo attachment, adhesion, and implantation. Using next-generation sequencing, we identified 81 DEmiRs between equine endometrium during the pre-attachment period of pregnancy (day 26–28) and endometrium during the mid-luteal phase of the estrous cycle (day 10–12). The identified DEmiRs appear to have a significant role in regulating the expression of genes that influence cell fate and properties, as well as endometrial receptivity formation. These miRNAs include eca-miR-21, eca-miR-126-3p, eca-miR-145, eca-miR-451, eca-miR-491-5p, members of the miR-200 family, and the miRNA-17-92 cluster. The target genes predicted for the identified DEmiRs are associated with ion channel activity and sphingolipid metabolism. Furthermore, it was noted that the expression of mucin 1 and leukemia inhibitory factor, genes potentially regulated by the identified DEmiRs, was up-regulated at day 26–28 of pregnancy. This suggests that miRNAs may play a role in regulating specific genes to create a favorable uterine environment that is necessary for proper attachment, adhesion, and implantation of the embryo in mares.

Sleep deprivation from mid-gestation leads to impaired of motor coordination in young offspring mice with microglia activation in the cerebellar vermis

ObjectiveTo investigate the effects of mid-pregnancy sleep deprivation (SD) in C57BL/6 J mice on the motor coordination of the offspring and to explore the potential mechanism of microglia activation in the cerebellar vermis of the offspring involved in the induction of impaired motor coordination development. MethodsC57BL/6 J pregnant mice were randomly divided into the SD and control groups. SD was implemented by the multi-platform method from first day of the middle pregnancy (gestation day 8, GD8). At postnatal day 21 (PND21), we measured the development of motor behavior and collected cerebellar vermis tissues to observe the activation of microglia by H&E staining, the expression of microglia-specific markers ionized calcium-binding adaptor molecule-1 (Iba-1) and cluster of differentiation 68 (CD68) by immunohistochemical, and interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-10 (IL-10), and tumor necrosis factor -α (TNF-α) by real-time quantitative PCR (RT-qPCR). ResultsIn the offspring of SD group, comparing to the control group, the total time of passage and the reverse crawl distance in the balance beam test, and the frequency of falls from the suspension cord was increased; with lower max rotational speed and shorter duration in the rotarod experiment. Further, we found that the microglia of cerebellar vermis tissues emerged an amoeba-like activation. The mean gray value of Iba-1 was lower, the density of positive cells of CD68 and the expression levels of IL-6 and TNF-α were increased. ConclusionsThe motor coordination of offspring is impaired, accompanying a SD from mid-pregnancy, and the cerebellar vermis showed microglia activation and pro-inflammatory response. It suggested the adverse effects of SD from mid-gestation on the development of motor coordination through the inflammatory response in the cerebellar vermis of the offspring.