Delivery Of Pups Research Articles

Generation of rat offspring from ovarian oocytes by xenotransplantation

The idea of utilizing unused oocytes present in the ovaries has been tested in various ways to produce offspring. However, only a limited number of studies succeeded in offspring generation. They include transplantation of ovaries into autologous or allogeneic animals, and acquisition of pups from oocytes obtained by transplanting mouse ovaries into immunodeficient rats. Here we report successful production of rat oocytes by transplanting rat ovaries under the kidney capsule of immunodeficient mice with addition of hormone administration to the mice. In addition, these oocytes were developed by in vitro fertilization, and transplanted into the oviducts of pseudopregnant rats, resulting in successful delivery of pups. The modified gene of the donor rat was confirmed to be correctly inherited to the pups. These results show that xenotransplantation of ovarian tissue makes it possible to leave offspring, beginning a new phase in developmental engineering.

Therapeutic Potential of Mesenchymal Stem Cell-Derived Extracellular Vesicles to Treat PCOS.

Polycystic ovary syndrome (PCOS) is known as the most common endocrine disorder in women. Previously, we suggested that human mesenchymal stem cells (MSCs) can reverse the PCOS condition by secreting factors. Here, we evaluated the therapeutic capability of MSC-derived extracellular vesicles (EVs), also known as exosomes, in both in vitro and in vivo PCOS models. Exosomes were used to treat androgen-producing H293R cells and injected in a mouse model through intraovarian and intravenous injection into a letrozole (LTZ)-induced PCOS mouse model. We assessed the effects of the exosomes on androgen-producing cells or the PCOS mouse model by analyzing steroidogenic gene expression (quantitative real-time polymerase chain reaction (qRT-PCR)), body weight change, serum hormone levels, and fertility by pup delivery. Our data show the therapeutic effect of MSC-derived EVs for reversing PCOS conditions, including fertility issues. Interestingly, intravenous injection was more effective for serum glucose regulation, and an intraovarian injection was more effective for ovary restoration. Our study suggests that MSC-derived exosomes can be promising biopharmaceutics for treating PCOS conditions as a novel therapeutic option. Despite the fact that we need more validation in human patients, we may evaluate this novel treatment option for PCOS with the following clinical trials.

Interaction between buprenorphine and norbuprenorphine in neonatal opioid withdrawal syndrome

Buprenorphine (BUP) is the preferred treatment for opioid use disorder during pregnancy but can cause neonatal opioid withdrawal syndrome (NOWS). Norbuprenorphine (NorBUP), an active metabolite of BUP, is implicated in BUP-associated NOWS. We hypothesized that BUP, a low-efficacy agonist of mu opioid receptors, will not antagonize NorBUP, a high-efficacy agonist of mu opioid receptors, in producing NOWS. To test this hypothesis, we treated pregnant Long-Evans rats with BUP (0, 0.01, 0.1 or 1mg/kg/day) ± NorBUP (1mg/kg/day) from gestation day 9 until pup delivery, and tested pups for opioid dependence using our established NOWS model. We used LC-MS-MS to quantify brain concentrations of BUP, NorBUP, and their glucuronide conjugates. BUP had little effect on NorBUP-induced NOWS, with the exception of 1mg/kg/day BUP significantly increasing NorBUP-induced NOWS by 58% in females. BUP and NorBUP brain concentrations predicted NOWS in multiple linear regression models. Interestingly, NorBUP contributed more to NOWS in females (βNorBUP = 51.34, p = 0.0001) than in males (βNorBUP = 19.21, P = 0.093), while BUP was similar for females (βBUP = 10.62, P = 0.0017) and males (βBUP = 11.38, P = 0.009). We are the first to report that NorBUP induces NOWS in the presence of BUP and it is more influential in females than males in the contribution of NorBUP to BUP-associated NOWS. These findings suggest that females are more susceptible to NorBUP-induced NOWS, and that treatment strategies that reduce prenatal NorBUP exposure may be more effective for females than males.

Preterm birth leads to a decreased number of differentiated podocytes and accelerated podocyte differentiation.

Preterm birth was previously identified as a high-risk factor for the long-term development of chronic kidney disease. However, the detailed pattern of podocyte (PD) changes caused by preterm birth and the potential mechanism underlying this process have not been well clarified. In present study, a rat model of preterm birth was established by delivery of pups 2days early and podometric methods were applied to identify the changes in PDs number caused by preterm birth. In addition, single-cell RNA sequencing (scRNA-seq) and subsequent bioinformatic analysis were performed in the preterm rat kidney to explore the possible mechanism caused by preterm birth. As results, when the kidney completely finished nephrogenesis at the age of 3weeks, a reduction in the total number of differentiated PDs in kidney sections was detected. In addition, 20 distinct clusters and 12 different cell types were identified after scRNA-seq in preterm rats (postnatal day 2) and full-term rats (postnatal day 0). The numbers of PDs and most types of inherent kidney cells were decreased in the preterm birth model. In addition, 177 genes were upregulated while 82 genes were downregulated in the PDs of full-term rats compared with those of preterm rats. Further functional GO analysis revealed that ribosome-related genes were enriched in PDs from full-term rats, and kidney development-related genes were enriched in PDs from preterm rats. Moreover, known PD-specific and PD precursor genes were highly expressed in PDs from preterm rats, and pseudotemporal analysis showed that PDs were present earlier in preterm rats than in full-term rats. In conclusion, the present study showed that preterm birth could cause a reduction in the number of differentiated PDs and accelerate the differentiation of PDs.

The impact of maternal obesity on in vivo uterine contractile activity during parturition in the rat.

Maternal obesity is associated with increased risk of prolonged and dysfunctional labor and emergency caesarean section. To elucidate the mechanisms behind the associated uterine dystocia, a translational animal model is required. Our previous work identified that exposure to a high-fat, high-cholesterol (HFHC) diet to induce obesity down-regulates uterine contractile associated protein expression and causes asynchronous contractions ex vivo. This study aims to investigate the impact of maternal obesity on uterine contractile function in vivo using intrauterine telemetry surgery. Virgin female Wistar rats were fed either a control (CON, n=6) or HFHC (n=6) diet for 6 weeks prior to conception, and throughout pregnancy. On Day 9 of gestation, a pressure-sensitive catheter was surgically implanted aseptically within the gravid uterus. Following 5 days recovery, intrauterine pressure (IUP) was recorded continuously until delivery of the 5th pup (Day 22). HFHC induced obesity led to a significant 1.5-fold increase in IUP (p=0.026) and fivefold increase in frequency of contractions (p=0.013) relative to CON. Determination of the time of labor onset identified that HFHC rats IUP (p=0.046) increased significantly 8h prior to 5th pup delivery, which contrasts to CON with no significant increase. Myometrial contractile frequency in HFHC rats significantly increased 12 h prior to delivery of the 5th pup (p=0.023) compared to only 3h in CON, providing evidence that labor in HFHC rats was prolonged by 9h. In conclusion, we have established a translational rat model that will allow us to unravel the mechanism behind uterine dystocia associated with maternal obesity.

17BIPHE2, an engineered cathelicidin antimicrobial peptide with low susceptibility to proteases, is an effective spermicide and microbicide against Neisseria gonorrhoeae.

Is 17BIPHE2, an engineered cathelicidin antimicrobial peptide with low susceptibility to proteases, a better spermicide in cervicovaginal fluid (CVF) than its parental peptides, LL-37 and GF-17? At the same mass concentration, 17BIPHE2 exhibited the highest spermicidal activity on human sperm resuspended in CVF-containing medium. LL-37 and its truncated peptide GF-17 exert both spermicidal and microbicidal activities, although they are prone to proteolytic degradation in body fluids. Spermicidal activities of 17BIPHE2 were evaluated in vitro in mouse and human sperm, both resuspended in medium, and then on human sperm incubated in CVF-containing medium; in the latter condition, the spermicidal activity and peptide stability in CVF of 17BIPHE2 were compared with that of LL-37 and GF-17. The in vivo contraceptive effects of 17BIPHE2 and the reversibility thereof were then assessed in mice. Finally, in vitro microbicidal effects of 17BIPHE2 on Neisseria gonorrhoeae were determined. Sperm motility and plasma membrane integrity were assessed by videomicroscopy and exclusion of Sytox Green, a membrane-impermeable fluorescent dye, respectively. Successful in vitro fertilization (IVF) was determined by the presence of two pronuclei in oocytes following their coincubation with capacitated untreated or 17BIPHE2-treated sperm. Sperm alone or with 17BIPHE2 were transcervically injected into female mice and successful in vivo fertilization was indicated by the formation of two-cell embryos 42-h postinjection, and by pregnancy through pup delivery 21-25 days afterwards. Peptide intactness was assessed by immunoblotting and HPLC. Reversibility of the contraceptive effects of 17BIPHE2 was evaluated by resumption of pregnancy of the female mice, pretranscervically injected with 17BIPHE2, following natural mating with fertile males. Minimum inhibitory/bactericidal concentrations of 17BIPHE2 on N. gonorrhoeae were obtained through microdilution broth assay. At the same mass concentration, 17BIPHE2 was a more effective spermicide than LL-37 or GF-17 on human sperm resuspended in CVF-containing medium, with the spermicidal concentration of 32.4 µM. This was mainly due to lower susceptibility of 17BIPHE2 to CVF proteases. Importantly, the reproductive tract of mouse females treated three times with 32.4 µM 17BIPHE2 remained normal and their fecundity resumed after stopping 17BIPHE2 treatment. For ethical reasons, the inhibitory effects of 17BIPHE2 on fertilization and pregnancy cannot presently be performed in women. Also, while our study has proven the effectiveness of 17BIPHE2 as a spermicide for mouse and human sperm in vitro, dosage formulation (e.g. in hydrogel) of 17BIPHE2 still needs to be developed to allow 17BIPHE2 to remain in the vagina/uterine cavity with controlled release for its spermicidal action. Since 17BIPHE2 also exerted bactericidal activity against N. gonorrhoeae at its spermicidal concentration, it is a promising candidate to be developed into a vaginal multipurpose prevention technology agent, thus empowering women against unplanned pregnancies and sexually transmitted infections. This work was supported by the Canadian Institutes of Health Research (PJT 173268 to N.T.). There are no competing interests to declare. N/A.

Synergistic interactions of sperm impairing bacteria: Impact on pregnancy outcome in mouse model

Earlier in our laboratory, the role of various individual sperm impairing microorganisms on sperm parameters and female infertility has been elucidated at higher doses. As, multiple bacterial species tend to exert more pathogenic effect in comparison to single organism hence, present study was carried out to evaluate that if consortia of these sperm impairing organisms can lead to infertility in female mice at sub fertility dose. For this, impact of individual bacterial strains of Escherichia coli, Pseudomonas aeruginosa, Pseudomonas aeruginosa, Klebsiella pneumoniae and consortia of Escherichia coli and Pseudomonas aeruginosa, Escherichia coli and Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae was examined on the motility, viability of mouse spermatozoa and fertility outcome. The results showed that the individual bacterial strains of E. coli, S. marcescens and K. pneumoniae could led to immobilization of spermatozoa by agglutination and P. aeruginosa led to immobilization of spermatozoa without agglutination. Also, all of them led to 100% sperm death in 45 min of incubation. In case of consortia of bacterial strains, the results showed sperm agglutination in all the cases and they were able to induce 100% sperm death at 30 min of incubation time. Further, in vivo studies were carried out to evaluate the impact of individual bacterial strains and consortia of bacterial strains on the fertility outcome in female Balb/c mice. For this, female mice were administered intravaginally with 101 cfu/20µl of individual bacterial strains or consortia of strains for 10 consecutive days or PBS. The results showed that both individual bacterial strains and consortia of bacterial strains were able to efficiently colonize the mouse vagina. Further, control group receiving phosphate buffer saline and groups receiving individual bacterial strains showed all the pregnancy related changes viz. abdominal distension, string of pearls on palpation as well as delivery of pups on completion of gestation period and delivery of pups. The histological examination of reproductive organs viz. uterus and ovary, of the female mice receiving PBS or individual bacterial strains showed the formation of corpus luteum in the ovary and the formation of decidua’s in the uterus, indicative of pregnancy. However, mice receiving consortia of bacterial strains did not show any pregnancy related changes throughout the experiment. Thus, these results indicate that the presence of consortia of sperm impairing microorganisms in vaginal milieu is efficient in provoking infertility even at subfertility doses.

Gamma aminobutyric acid signaling disturbances and altered astrocytic morphology associated with Bisphenol A induced cognitive impairments in rat offspring.

Bisphenol A (BPA) is a well-recognized endocrine disruptor and is globally used in the manufacture of many plastic items. Multiple studies suggest links between prenatal BPA exposure and alterations in neurodevelopment and behaviors in children, even at lower levels. This study was conducted to reveal the role of astrocyte morphology and Gamma aminobutyric acid (GABA) signaling in BPA induced cognitive defects in the offspring of Wistar albino rats when exposed during the prenatal and postnatal periods. Dams of Wistar albino rats were exposed to a dose of 5mg/kg body weight of BPA throughout the pregnancy and lactation period until the third postnatal day (PND). After delivery of pups, cognitive tests were carried out on the 21st, 24th, and 28th PNDs. Blood samples were collected for measurement of serum GABA levels. On the same day as the blood collections, pups were sacrificed and their right frontal cortices were dissected out. Immunohistochemical analysis for glial fibrillar acidic protein + astrocytes was conducted. Pre and postnatal BPA exposure led to anxiety like behavior in pups. This exposure also resulted in reduced serum GABA concentrations. Immunohistochemical analysis revealed reduced astrocyte numbers as well as decreased numbers of dendritic spines in the BPA exposed pups. BPA exposure during critical periods of development leads to cognitive impairments that correlate with the defects in the GABA signaling pathways and deteriorated morphology of the astrocytes in the offspring of the Wistar rats.

Vitamin A Status in Neonatal and Weanling Rats Reared by Mothers Consuming Normal and High Fat Diets with or Without Vitamin A Supplementation

Abstract Objectives Although the circulating level of vitamin A (VA) is reported to be significantly lower in obese adults, it is unknown if maternal adiposity would influence the VA status of the offspring. The aim of this study was to investigate the status and distribution of VA in the offspring of rats consuming a normal fat diet (NFD) or a high fat diet (HFD), with or without VA supplementation. Methods Sprague-Dawley rats arrived on their second day of gestation and were randomized to either a NFD (25% fat) or a HFD (50% fat) both with an adequate level of VA at 2.6 mg/kg. Upon the delivery of pups, half of rat mothers from the NFD or HFD cohort were switched to a NFD or a HFD both with supplemented VA at 129 mg/kg (NFD + VA and HFD + VA group), respectively. The other half remained on their diets with adequate VA (NFD and HFD group). At postnatal day 14 (P14) and P25, 4 pups/group were euthanized. At P25, the remaining weanling pups (n = 3/group) were fed diets of their respective mothers until being euthanized at P35. Concentrations of total retinol in serum, liver, visceral white AT (WAT), and brown AT (BAT) were measured by UPLC. Results Through the study, pups in non-supplemented groups showed a normal serum retinol concentration but a marginal liver VA status. At all the time points, serum retinol concentration was significantly (P < 0.05) increased in VA-supplemented groups as compared to non-supplemented ones; no significant difference was noted between the NFD and the HFD group. At P14, serum retinol concentration was significantly lower in the HFD + VA group than in the NFD + VA group. In liver, WAT, and BAT, both the concentration and the mass of total retinol was significantly higher in HFD + VA and NFD + VA group than in their respective control at all the time points, with no significant difference noted between the two control groups. At P25, both the concentration and the mass of total retinol in liver and in WAT was significantly higher in HFD + VA than in NFD + VA group. Conclusions Maternal VA supplementation added to both NFD and HFD raised VA status of the offspring. A lower VA concentration in serum but higher concentration in liver and WAT was observed in the HFD + VA group as compared to the NFD + VA group at given time points, indicating that maternal HFD consumption potentially compromised the availability of functional VA in circulation by increasing its deposition in storage organs. Funding Sources NIH.

Vitamin A Supplementation During Suckling and Post-Weaning Periods Improves Systemic and Adipose Tissue Redox Status in Offspring of Rats Consuming a High-Fat Diet

Abstract Objectives Dietary fat can contribute to oxidative stress and adipose tissue (AT) accrual which exacerbates redox imbalance. Vitamin A (VA) has been shown to regulate AT development and associated metabolic conditions in adult models; however, such effects of VA in early life stages, especially on redox status, has barely been studied. The aim of this study was to assess the effects of VA supplementation during suckling and post-weaning periods on systemic and AT redox status in rats reared by mothers consuming a high-fat diet (HFD). Methods Sprague-Dawley rats arrived on their second day of gestation and were randomized to either a normal-fat diet (NFD = 25% fat) or a HFD (50% fat) both with adequate VA at 2.6 mg/kg. Upon delivery of pups, half of rat mothers from the NFD cohort or HFD cohort were switched to a NFD or a HFD both with supplemented VA at 129 mg/kg (NFD + VA and HFD + VA group), respectively. The other half of mothers remained on their diets with adequate VA (NFD and HFD group). At postnatal day 14 (P14), P25, and P35, pups (n = 4 or 3/group/time point) were euthanized. Concentrations of VA, lipid peroxides, and antioxidant capacity (AC) of serum, visceral white AT (WAT) and brown AT (BAT) were measured. Results At all three time points, the lipid peroxides concentration in WAT was significantly lower in VA-supplemented groups as compared to their respective control (P < 0.0001 for NFD + VA vs. NFD; P < 0.05 for HFD + VA vs. HFD); lipid peroxides were significantly (P < 0.01) higher in the HFD + VA group than in the NFD + VA group. A significantly (P < 0.05) positive correlation between the amount of VA and the lipophilic AC of the WAT was noted. For BAT, the NFD + VA group showed a significantly (P < 0.001) lower lipid peroxides concentration as compared to the NFD group. However, such effect was not observed in the HFD cohort. Serum lipid peroxides in the HFD + VA group were significantly (P < 0.05) lower than in the HFD group. A significant (P < 0.0001) negative correlation was found between the serum concentration of VA and lipid peroxides. Conclusions VA supplementation during suckling and post-weaning period improved both the systemic and WAT redox status of rats reared by mothers consuming a HFD, as well as the status within both WAT and BAT of pups nursed by mothers consuming a NFD. The differing results in WAT and BAT may suggest their different roles in redox biology. Funding Sources NIH

Exposure to carbon black nanoparticles during pregnancy persistently damages the cerebrovascular function in female mice

Maternal exposure to carbon black nanoparticles (CBNPs) during pregnancy have been well documented to induce harmful outcomes of offspring on brain function. However, it remains largely unknown whether females exposed to CBNPs during sensitive period of pregnancy can cause the neurotoxic effects on their own body after parturition. In this study, our results showed that pregnancy CBNPs exposure induced the persistent pathological changes in the cerebral cortex tissues and impaired cerebrovascular function of mice manifested by significant alterations of endothelin-1, endothelial nitric oxide synthase, vascular endothelial growth factor-A and ATP-binding cassette transporter G1. Intriguingly, we observed that these deleterious effects on brain and cerebrovascular functions in mice could persist for 49 days after delivery of pups. By using in vitro human umbilical vein endothelial cells, we further verified the potential vascular dysfunction after CBNPs exposure. In summary, our results provide the first evidence that pregnancy CBNPs exposure-induced brain pathological changes and cerebrovascular dysfunction can persist for a relative long time. These finding suggest exposure to CBNPs during sensitive stages of pregnancy may not only show the harmful effects on offspring neurodevelopment, but also result in the irreversible brain damage on mother body.

Role of maternal folic acid on implantation of embryos and on maternal body weight & offspring's birth weight

Introduction There has been a much debate on the role of pre-/periconceptional and prenatal folic acid (FA) supplements for the long-term health of their offspring. Maternal deficiency of FA or excess of FA would likely to affect the developing fetus at critical stages of development to cause long-lasting health consequences in their offspring. The present study aimed to know the effect of folic acid supplementation and folic acid absence diet during pregnancy on the number of implantation, endometrial changes at the implantation site and on maternal body weight & offspring's birth weight. Material & Methods Thirty-Six Wistar strained female rats were approved for experimental use by the Institutional Animal Ethical Committee. Rats were randomly divided into two groups with 18 rats in each group. Further, each group has been divided into three sets with 6 rats in each. The control set was fed a normal diet, the second set was fed a diet containing no FA. 1% Succinyl sulfathiazole was added to the diet with no FA content to reduce/ inhibit the gut flora responsible for the synthesis of FA in their body. The third set was given diet with an excess of FA (40mg/kg body weight/day). Rats were given respective diet for 4 weeks before mating, continued throughout their pregnancy and 5 weeks after the delivery of pups (weening period). All female rats were paired with males of their own strain overnight at room temperature (25°C). To check whether they mated or not, the vaginal smear was taken in the early morning and stained with methylene blue and confirmed the presence of sperm plug under microscopy. On confirmation of sperm plug, the gestational day was termed as day 1 (GD1). Rats have remained in their cage and diets until they were sacrificed during gestation from day GD6-GD9. Rats were remained in their cage and with their respective diets until they delivered the pups. Delivered pups were counted for each rat and gross malformation in each pup was noted. Immediately after the delivery, all the pups born to each rat were weighed individually in the digital weighing machine. Results Average number of pups born to pregnant dams fed with FA supplementation, FA absent diet, and in normal diet was found to be 8,8, and 9 respectively. Regarding the birth weight of pups born to pregnant dams fed with FA supplementation the average weight was found to be 6.01-5.09 gm, with evidence diet it was 5.51-5.08 gm, and least weight was observed in pups born with FA absent diet (4.74-3.31 gm). Pair-wise comparison was done to correlate the significant association between the different groups. It showed significant difference when compared with pups born to FA supplementation diet to pups born to FA absent diet. Also, significant differences were noted when compared pups born to FA absent diet mother with pups born to FA supplementation and with pups born to normal diet. Conclusion Our study provides clear evidence that FA is essential in successful implantation by providing favorable receptive environment to receive the implantation-competent blastocyst for a successful pregnancy. Birth weight of pups born to pregnant dams fed with FA supplementation was found to be significantly increased when compared to those with the diet with normal amount of FA, and least weight wasobserved in pups born with FA absent diet. As the fetal birth weight is the best predictor of pregnancy outcome as the growth restriction has been associated with high morbidity and mortality. FA supplementation is essential during the period of pregnancy to prevent untoward effects on pregnancy-related complications in mother and to prevent birth defects in fetuses. Further research from longitudinal studies is warranted to confirm these results before its recommendation to the public health.

Prefrontal Cortical and Behavioral Adaptations to Surgical Delivery Mediated by Metabolic Principles.

We previously observed an association between mode of delivery and brain mitochondrial mechanisms in pups. We also showed that mitochondrial processes impact adult behavior. However, no experimental data is available to causally connect mode of delivery with cellular processes of neurons in the cerebral cortex and adult behavior. Here we show that surgical delivery of pups alters mitochondrial dynamics and spine synapses of layer 3 pyramidal neurons of the prefrontal cortex compared to the values of mice delivered vaginally. These alterations in ultrastructure seen in adult mice delivered surgically were associated with the development of behavioral phenotypes resembling those characteristic of animal models of psychiatric illness. This included impaired performance in prepulse inhibition as well as hyperlocomotion in the open field and elevated plus maze tests. Knocking out a mitochondria-related gene, UCP-2, blocked cellular and behavioral adaptations induced by surgical delivery. These results highlight a crucial role for brain mitochondrial adaptations in the process of birth to affect neuronal circuitry in support of normal and altered adult behaviors. Further, these findings were supported with neuroimaging data from human neonates delivered vaginally and surgically, suggesting that the murine findings have human clinical relevance.

Pup-Associated Conditioned Place Preference Reaction and Maternal Care in Depressive WAG/Rij Rats

The acquisition of a conditioned place preference reaction (CPPR) associated with own pups and foster pups was compared in female WAG/Rij and Wistar rats, along with maternal behavior in a “place preference” chamber from day 2 to day 8 after birth. In addition, females’ behavior was assessed in the open field, elevated plus maze, and forced swimming tests before pregnancy and after birth of pups. Female WAG/Rij rats acquired the CPPR associated with their own pups (WAG/Rij) and foster pups (Wistar) less well. Thus, the number of females with increases in the time spent in the initial non-preferred sector after it was combined with pups and the number of females achieving the CPPR acquisition criterion were smaller in WAG/Rij rats than Wistar rats. Female WAG/Rij rats, as compared with female Wistar rats, showed lower levels of maternal care in the place preference chamber in relation to both their own pups and foster pups: the numbers of approaches to the pups were smaller, as were the numbers of retrievals, and the non-feeding-related times spent in contact with pups were shorter. The durations of immobility in the forced swimming test were greater in WAG/Rij females than Wistar females both before pregnancy and after delivery of pups, which is evidence for a stable depression-like state. Before pregnancy, there were no statistically significant interstrain differences in anxiety levels. After birth of pups, anxiety levels in WAG/Rij females decreased, while those in Wistar females showed no significant change. These results provide evidence that the worse acquisition of the CPPR and the decreased maternal care in depressed WAG/Rij females were not associated with the features of their own pups but were related to their depressive state. In other words, pups provided weaker reinforcement for depressed mothers than for “normal” (nondepressed) mothers.

EFFECTS OF UNANI ANXIOLYTICS (SOMINA) ON GENERAL REPRODUCTIVE PERFORMANCE AND TERATOLOGY IN RATS

Objective: Somina (herbal medicine) is used in Pakistan as Unani anxiolytics. It is composed of five medicinal plants. The current work was designed to evaluate the general reproductive and teratogenic effects of somina in two consecutive generations of rats according to the OECD guideline.Methods: Fertility study (a two-phase study) was done in Sprague-Dawley rats. 1st part: three groups’ female rats (10 rats each group) received different doses orally. First group: The control group (saline), a single oral human dose of somina (2nd group: 285 mg/kg/day) and the high dose of somina (3rd group: 1g/kg/day) during the whole period of gestation till the delivery of pups named as F1 Breed. For the second part of study ten females were selected from each F1 breed (control, somina 285 mg/kg/d, somina 1g/kg/day) and administered the same treatment from day first of mating than the entire period of gestation until F1 breed delivered pups (F2 breed). For F1 and F2 breed the fertility index and litter size were determined. Some of the female rats (F1 and F2) were anesthetized and autopsied. The blood sample was subjected to biochemical analysis and serum liver function test: bilirubin, gamma-glutamyl transferase (γGT), alanine aminotransferase (ALT: SGPT), aspartate aminotransferase (AST: SGOT) and alkaline phosphatase (ALP) were measured spectrophotometrically. The uterine growth index, fertility index, and litter size were also measured to evaluate the teratogenic effects of somina treated rats.Results: The data showed that any significant different (P>0.05) was not found during the maternal examination (uterine growth index, fertility index) and reproductive parameters (litter size, the quantity of fetus, aborted or absorbed fetus) in somina treated rats as compare to control rats (P>0.05). Control and treated Pups did not show any significant (P>0.05) malformation and any congenital defects. Non-significant (P>0.05) changes were observed in liver function test. It was found normal in all groups. Macroscopic autopsy examination also did not reveal any significant (P>0.05) pathological findings in the liver, kidneys, and uterus.Conclusion: The oral administration of somina during the gestational period of pregnant female rats was not teratogenic/fetotoxic. Any adverse or deleterious effects were not observed at the dose of 285 mg/kg (human dose) or 1g/kg (3times greater than the human dose) during pregnancy, and it is safe in rats.

Pregnancy primes anti-fetal T cell memory capable of allograft destruction (TRAN3P.874)

Abstract Pregnancy-induced anti-fetal antibody can destroy allografts and excludes many women from transplantation, but anti-fetal T cell responses are poorly understood and are not currently evaluated in potential organ recipients. To test whether pregnancy (PG) primes graft-reactive T cell memory, we compared Ova skin graft survival (GS) between naive B6 mice (N), mice mated to Ova transgenic (tg) males (Ova PG-sensitized; OP), or mice sensitized by a previous Ova skin graft (OS). Skin grafts were placed 30 days after either Ova pup delivery (OP group) or after rejection of an Ova skin graft (OS group). OP females rapidly rejected Ova skin grafts with second set kinetics identical to OS females [GS (days): 11.3±1.5(OP) vs. 10.3±0.6(OS) vs. 15±0.7(N); p<.01]. To test the effects on antigen-specific T cells, mice were injected with Ova-specific TCR tg T cells. Flow cytometric analysis 30 days after pup delivery revealed memory T cell differentiation in response to fetal antigen [%anti-Ova CD8+CD44+CD62L-: 85%(Ova-mated) vs. 38%(Balb/c-mated)]. OP mice also produced enhanced effector T cells upon secondary challenge with an Ova skin graft [%anti-Ova CD8+IFNγ+: 60%(OP) vs. 43%(Balb-mated) vs. 35%(N)]. These results indicate that PG can prime T cell memory that gives rise to secondary effector T cells that mediate rapid graft destruction. Stimulation of PG-induced anti-fetal T cell memory may contribute to the inferior GS observed in women who receive a transplant from a child or spouse.

1384 THE ROLE OF SIMULATED HUMAN CHILDBIRTH INJURIES IN A LYSYL OXIDASE LIKE-1 KNOCKOUT MOUSE MODEL OF PELVIC ORGAN PROLAPSE

You have accessJournal of UrologyUrodynamics/Incontinence/Female Urology: Pelvic Prolapse1 Apr 20101384 THE ROLE OF SIMULATED HUMAN CHILDBIRTH INJURIES IN A LYSYL OXIDASE LIKE-1 KNOCKOUT MOUSE MODEL OF PELVIC ORGAN PROLAPSE Andrew Lenis, Bruce Kinley, Dan Li Lin, and Margot Damaser Andrew LenisAndrew Lenis More articles by this author , Bruce KinleyBruce Kinley More articles by this author , Dan Li LinDan Li Lin More articles by this author , and Margot DamaserMargot Damaser More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2010.02.1034AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Pelvic organ prolapse (POP) is one of several female pelvic floor disorders, a group of debilitating and poorly understood conditions that may affect up to half of all women in the U.S. No animal besides human routinely develops POP. Dysfunctional elastin homeostasis, in part regulated by the enzyme lysyl oxidase like-1 (LOXL1), has been implicated in the pathophysiology of POP in humans. LOXL1 knockout (KO) mice display anatomical changes similar to those in POP after delivery of mouse pups. We hypothesized that simulated human childbirth injury by vaginal distension (VD) in LOXL1 KO mice would increase the incidence of POP compared to vaginal delivery or Caesarean delivery of pups. METHODS Age-matched, nulliparous LOXL1 KO mice were randomized into 6 groups and allowed to breed at 6 weeks old. Two groups were delivered by Caesarean section; two groups were allowed to deliver vaginally; and two groups of nulliparous mice were used as controls. One group from each of these delivery modes underwent VD. The other group underwent sham VD. Weekly assessment of POP using the validated Mouse Pelvic Organ Prolapse Quantification (MOPQ) staging system was used to determine timing and staging of POP for 12 consecutive weeks after injury. RESULTS Twelve mice delivered vaginally, 13 mice delivered via Caesarean section, and 4 nulliparous mice were used as controls. Six of the 12 mice that delivered vaginally underwent VD and 6 underwent sham VD. Nine of the 13 mice that delivered via Caesarean section underwent VD and 4 underwent sham VD. Delivery groups took significantly less time to develop POP (3.7 ± 0.8 weeks) than Caesarean section groups (5.9 ± 0.7 weeks; Wilcoxon, p=0.04). Time to POP after VD was not significantly different than after sham VD, regardless of delivery mode. Nulliparous mice rarely develop POP, even after VD. CONCLUSIONS Our results indicate that simulated human birth injury does not decrease the time to POP in LOXL1 KO mice, suggesting that hormonal changes and aspects of delivery other than direct trauma play an important role in POP development. The mechanism of these findings will be investigated in future studies. Cleveland, OH© 2010 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 183Issue 4SApril 2010Page: e535 Advertisement Copyright & Permissions© 2010 by American Urological Association Education and Research, Inc.MetricsAuthor Information Andrew Lenis More articles by this author Bruce Kinley More articles by this author Dan Li Lin More articles by this author Margot Damaser More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Preterm and Term Cervical Ripening in CD1 Mice (Mus musculus): Similar or Divergent Molecular Mechanisms?1

Premature cervical ripening is believed to contribute to preterm birth (PTB). Preterm cervical ripening may be due to an aberrant regulation in timing of the same processes that occur at term, or may result from unique molecular mechanisms. Using mouse models of PTB, this study sought to investigate if the molecular mechanisms that govern cervical ripening were similar between preterm and term. Lipopolysaccharide (LPS) is infused into the uterine horn to create a mouse model of inflammation-induced PTB. For a noninfectious model of PTB, RU486 was administered. Both models result in delivery of pups in 8-24 h. Cervical tissues were collected from these models, as well as throughout gestation. Cervical tissues from E15 (preterm), E15 LPS (preterm inflammation), and E18.5 (term) were used for microarray analysis (n = 18). Additional experiments using gestational time course specimens were performed to confirm microarray results. Specific gene pathways were differentially expressed between the groups. Genes involved in immunity and inflammation were increased in the cervix in inflammation-induced PTB; term labor was not associated with differential expression of immune pathways. Cytokine expression was not increased in cervices during term labor, but was increased in the pospartum period. Epithelial cell differentiation pathway was significantly altered in term, but not preterm, labor. Activation of immune pathways may be sufficient for cervical ripening, but does not appear necessary. Differential expression of the epithelial cell differentiation pathway appears necessary in the process of cervical repair. Our results indicate that the molecular mechanisms governing preterm and term cervical ripening are distinctly different.

Successful Delivery of Pups from Cryopreserved Canine Embryos.

Successful Delivery of Pups from Cryopreserved Canine Embryos.

Effets de l'envenimation par Buthus occitanus tunetanus sur un modèle expérimental murin de gestation

Scorpion envenoming is less studied in pregnant victims. In this work, the effect of Buthus occitanus tunetanus on parturition in late pregnancy was studied in an animal model. Four groups of six primigravid female rats, each one at the 22 nd day of pregnancy, were used. The first two groups had received an intra-peritoneal injection of 500 μg/kg of Buthus occitanus tunetanus crude venom or a physiological saline solution and left until foetal delivery. Then, the time elapsed until the first pup delivery and that separating the first and latest ones were measured. The other two groups served for the uterine electrophysiological activity exploration. Rats were anaesthetized, artificially ventilated and had received an intraperitoneal injection of 500 μg/kg of Buthus occitanus tunetanus crude venom or a physiological saline solution. Our results showed a significant increase of the latency to foetal delivery, labour time, and uterine contractile activity in envenomed rats compared to controls. Such signs are usually seen in dynamic dystocia. It was concluded that Buthus occitanus tunetanus envenoming might induce a dynamic dystocia, when it occurred in late pregnancy. To cite this article: H. Ben Nasr et al., C. R. Biologies 330 (2007).