Contralateral Horn Research Articles

Transuterine relocation of pregnant uterine horn segment in an exploratory rat model with implications for tubal ectopic pregnancy

Ectopic pregnancy affects ~ 2% of pregnancies annually in the United States, with no current treatments allowing for the continuation of the pregnancy. Thus, this study sought to initiate an investigation into the potential design of a surgical technique, in an animal model, that could serve as a foundation for future research into the potential of relocating an ectopic embryo into the uterus at the human level. Female Long-Evans rats were randomly assigned to one of two groups: Embryo Relocation (ER; n = 12; underwent embryo relocation surgery) and Normal Pregnancy (NP; n = 12; carried a normal pregnancy; no surgery). Eight rats/group were allowed to carry their pregnancy to term and deliver, while four had their uteri collected at the end of gestation. Briefly, for the ER group, a uterine horn containing 1–2 embryos was translocated to the contralateral horn, which had been incised and cleared of its contents, prior to being wrapped around the relocated horn. Rat weight, food consumption and vaginal impedance of the mothers were measured throughout the experiment. Ultrasounds were performed and fetal heart rates measured on day 20–21 of gestation. Additionally, rat weight of all offspring was measured at adulthood. Our findings indicate that, in the ER group, 15/15 (100%) of the relocated embryos had detectable heart rates at the end of gestation (within the normal range), 14/15 (93%) were delivered vaginally, and 12/14 (86%) survived until adulthood. A significant decrease in rat weight and food consumption was observed only on the day following surgery. Fertility, as measured by vaginal impedance, was minimally impacted by surgery. Moreover, there was no significant difference between groups in average body weight of offspring at adulthood. Histological analysis indicated a thicker placenta in the ER group, attributable to the fetal part of the placenta, potentially indicating compensatory mechanisms. Our findings reflect a successful transuterine embryo relocation followed by vaginal birth and survival of offspring to adulthood, in a rat model. Such findings lay the foundation for future preclinical research in higher animals, with potential implications on a procedure relevant to human ectopic embryo relocation.

Long-term pulmonary and neurodevelopmental impairment in a fetal growth restriction rabbit model.

Fetal growth restriction (FGR) remains one of the main obstetrical problems worldwide, with consequences beyond perinatal life. Animal models with developmental and structural similarities to the human are essential to understand FGR long-term consequences and design novel therapeutic strategies aimed at preventing or ameliorating them. Herein, we described the long-term consequences of FGR in pulmonary function, structure, and gene expression, and characterized neurodevelopmental sequelae up to preadolescence in a rabbit model. FGR was induced at gestational day 25 by surgically reducing placental blood supply in one uterine horn, leaving the contralateral horn as internal control. Neonatal rabbits born near term were assigned to foster care in mixed groups until postnatal day (PND) 21. At that time, one group underwent pulmonary biomechanical testing followed by lung morphometry and gene expression analysis. A second group underwent longitudinal neurobehavioral assessment until PND 60 followed by brain harvesting for multiregional oligodendrocyte and microglia quantification. FGR was associated with impaired pulmonary function and lung development at PND 21. FGR rabbits had higher respiratory resistance and altered parenchymal biomechanical properties in the lungs. FGR lungs presented thicker alveolar septal walls and reduced alveolar space. Furthermore, the airway smooth muscle content was increased, and the tunica media of the intra-acinar pulmonary arteries was thicker. In addition, FGR was associated with anxiety-like behavior, impaired memory and attention, and lower oligodendrocyte proportion in the frontal cortex and white matter. In conclusion, we documented and characterized the detrimental pulmonary function and structural changes after FGR, independent of prematurity, and beyond the neonatal period for the first time in the rabbit model, and describe the oligodendrocyte alteration in pre-adolescent rabbit brains. This characterization will allow researchers to develop and test therapies to treat FGR and prevent its sequelae.

Early pain experiences in dairy calves on pain sensitivity later in life

In humans, early painful experiences can increase pain sensitivity later in life, but little is known regarding this phenomenon in cattle. This study assessed if a painful event early in life affects later pain sensitivity in 40-d-old calves. Holstein calves (n = 26) were randomly assigned to control or treatment conditions, blocked within sex and birth weight. At 9.5 ± 1.8 d old, treatment calves had 1 horn bud disbudded using caustic paste, whereas control calves had 1 horn sham disbudded; in both cases, multimodal pain control was provided. All calves had the contralateral horn bud disbudded using a hot iron 4 wk later, again with multimodal pain control. Mechanical nociceptive responses were assessed weekly using an algometer applied adjacent to both horn buds and on the rump, beginning 3 d before the first disbudding and ending 30 d after the second disbudding. Following the second disbudding, both groups of calves showed evidence of increased sensitivity (i.e., algometer pressure declined 3.69 ± 0.60 √N to 2.13 ± 0.70 √N) on the contralateral bud, but there was no difference between the control group and the treatment groups. An interaction between treatment and time, likely driven by treatment differences, was found on the rump when tested 5 h after the second disbudding event. These responses are not consistent with the hypothesis that an early pain experience results in increased sensitivity to later painful experiences.

Bidirectional Feto-Maternal Traffic of Donor Mesenchymal Stem Cells Following Transamniotic Stem Cell Therapy (TRASCET)

PurposeTransamniotic stem cell therapy (TRASCET) with mesenchymal stem cells (MSCs) has emerged experimentally as a potential treatment for different congenital diseases and maternal diseases of pregnancy. The broad applicability of TRASCET is predicated on hematogenous routing of donor MSCs via the placenta. We investigated whether donor MSC kinetics includes bidirectional traffic between the fetus and mother. MethodsEight time-dated dams had their fetuses (n = 96) divided in 4 groups on gestational day 17 (E17, term = E21). Groups populating one uterine horn received intra-amniotic injections (50 μL) of either donor amniotic fluid-derived MSCs (2×106 cells/mL) labelled with a firefly luciferase reporter gene (MSC-injected, n = 32), or of acellular luciferase (luciferase-injected, n = 26). Contra-lateral (CL) horn fetuses received no injection (MSC-CL, n = 20 and luciferase-CL, n = 18). At term, samples from 11 fetal anatomical sites from CL fetuses, along with placentas from all fetuses and maternal blood were screened for luciferase activity via microplate luminometry. ResultsOverall survival was 95 % (91/96). When controlled by the acellular injection, positive luciferase activity was observed in the placentas of all MSC-injected fetuses, confirming viability of the donor cells at term. When controlled by the acellular injection group, MSC-CL fetuses showed positive luciferase activity in the bone marrow, peripheral blood, brain and skin (p = <0.001–0.048). No luciferase activity was detected in any maternal blood sample. ConclusionAmniotic fluid-derived MSCs can traffic between the fetus and mother in both directions after simple intra-amniotic injection, in a healthy rat model. This phenomenon must be considered in TRASCET performed in twin/multiple pregnancies. Level of evidenceN/A (animal and laboratory study).

Differences in Microbial Community Composition between Uterine Horns Ipsilateral and Contralateral to the Corpus Luteum in Beef Cows on Day 15 of the Estrous Cycle.

This study evaluated differences in uterine microbiota composition between uterine horns ipsilateral and contralateral to the corpus luteum of beef cows on day 15 of the estrous cycle. Cows (n = 23) were exposed to an estrus synchronization protocol to exogenously induce synchronized ovulation. Cows were then euthanized on day 15 of the estrous cycle, and individual swabs were collected from uterine horns ipsilateral and contralateral to the corpus luteum using aseptic techniques. DNA was extracted, and the entire (V1-V9 hypervariable regions) 16s rRNA gene was sequenced. Sequences were analyzed, and amplicon sequence variants (ASVs) were determined. Across all samples, 2 bacterial domains, 24 phyla, and 265 genera were identified. Butyribirio, Cutibacterium, BD7-11, Bacteroidales BS11 gut group, Ruminococcus, Bacteroidales RF16 group, and Clostridia UCG-014 differed in relative abundances between uterine horns. Rikenellaceae RC9 gut group, Bacteroidales UCG-001, Lachnospiraceae AC2044 group, Burkholderia-Caballeronia-Paraburkholderia, Psudobutyribibrio, and an unidentified genus of the family Chitinophagaceae and dgA-11 gut group differed between cows that expressed estrus and those that did not. The composition of the microbial community differed between the ipsilateral and contralateral horns and between cows that expressed estrus and cows that failed to express estrus, indicating that the uterine microbiota might play a role in cow fertility.

Laser ablation of the equine oviductal papilla as a novel contraceptive technique

Reproductive control of wild horses has frustrated land management groups for decades. GnRH and Zona Pellucida vaccines have a limited duration of efficacy, intrauterine devices are unpredictably retained, and attempts to ovariectomize mares in the western United States were met with public outcry in 2018. The development of a non-surgical permanent sterilization technique has the potential to revolutionize management of wild equids on public lands. The objectives of this study were to develop a safe and effective sterilization technique for reproductively healthy mares. We hypothesized that laser ablation of the oviductal papillae would be an effective method of permanent sterilization. Seven light-breed reproductively healthy mares (5-21yrs) were enrolled in the study. On enrollment, mares were sedated for hysteroscopy, and the endometrium and oviductal papillae were examined using a 103cm flexible endoscope (Olympus GIF-160 Gastroscope, Center Valley, PA) attached to an Olympus EVIS EXERA CV-160 video processor. In five mares, a 600 μm laser fiber was advanced through the biopsy channel and a diode laser (Dornier Medilas D, Dornier MedTech America, Inc., Kenneasaw, GA) was used to deliver direct contact pulses of 3 to 5 seconds in duration to the oviductal papilla. The endoscope was then guided up the contralateral horn and the process repeated. In two control mares, the oviductalpapillae were visualized, but no laser ablation was performed. All mares received 5 mg dinoprost IM and a uterine lavage within 4 hours post-procedure. Transrectal ultrasound was performed every 2-3 days until a 35mm follicle was detected, and then mares were bred using semen from a fertile stallion. Pregnancy status was determined by transrectal ultrasound 14 days post-ovulation, luteolysis was induced, and mares were rebred on 2-4 consecutive cycles. Control mares conceived on 6 out of 9 cycles (67% pregnancy rate). Pregnancy rate was significantly lower in the treatment group (5%, 1 out of 20 cycles, p=0.003). The first mare that was laser ablated conceived on the 4th cycle, and repeat hysteroscopy determined that the left oviduct was not effectively ablated. The third mare was diagnosed with peritonitis via abdominocentesis 6 hours post hysteroscopy. The mare was treated with broad spectrum antibiotics and recovered uneventfully. Three mares were re-bred two years after the procedure and did not conceive. In conclusion, when laser ablation of the oviductal papillae was appropriately performed, scar tissue formation effectively prevented pregnancy for a minimum of four months post-procedure. Laser ablation of the oviductal papillae is a promising technique for permanent sterilization of the mare and a follow-up long-term fertility study is warranted in a larger group of mares is warranted.

Lactoferrin/sialic acid prevents adverse effects of intrauterine growth restriction on neurite length: investigations in an in vitro rabbit neurosphere model.

Intrauterine growth restriction (IUGR) is a well-known cause of impaired neurodevelopment during life. In this study, we aimed to characterize alterations in neuronal development underlying IUGR and discover strategies to ameliorate adverse neurodevelopment effects by using a recently established rabbit in vitro neurosphere culture. IUGR was surgically induced in pregnant rabbits by ligation of placental vessels in one uterine horn, while the contralateral horn remained unaffected for normal growth (control). At this time point, rabbits were randomly assigned to receive either no treatment, docosahexaenoic acid (DHA), melatonin (MEL), or lactoferrin (LF) until c-section. Neurospheres consisting of neural progenitor cells were obtained from control and IUGR pup's whole brain and comparatively analyzed for the ability to differentiate into neurons, extend neurite length, and form dendritic branching or pre-synapses. We established for the very first time a protocol to cultivate control and IUGR rabbit neurospheres not only for 5 days but under long-term conditions up to 14 days under differentiation conditions. Additionally, an in vitro evaluation of these therapies was evaluated by exposing neurospheres from non-treated rabbits to DHA, MEL, and SA (sialic acid, which is the major lactoferrin compound) and by assessing the ability to differentiate neurons, extend neurite length, and form dendritic branching or pre-synapses. We revealed that IUGR significantly increased the neurite length after 5 days of cultivation in vitro, a result in good agreement with previous in vivo findings in IUGR rabbits presenting more complex dendritic arborization of neurons in the frontal cortex. MEL, DHA, and SA decreased the IUGR-induced length of primary dendrites in vitro, however, only SA was able to reduce the total neurite length to control level in IUGR neurospheres. After prenatal in vivo administration of SAs parent compound LF with subsequent evaluation in vitro, LF was able to prevent abnormal neurite extension. We established for the first time the maintenance of the rabbit neurosphere culture for 14 days under differentiation conditions with increasing complexity of neuronal length and branching up to pre-synaptic formation. From the therapies tested, LF or its major compound, SA, prevents abnormal neurite extension and was therefore identified as the most promising therapy against IUGR-induced changes in neuronal development.

Highly Effective Frontal Stunning Procedure Using a Pneumatic Penetrating Captive Bolt in Water Buffaloes

An effective frontal stunning procedure in water buffaloes was assessed using a pneumatic penetrating captive bolt (PPCB) with high air pressure. The study contemplated two phases; first, 352 buffaloes and 168 post-mortem heads were evaluated to determine the most effective anatomical site for stunning. Then, the second phase (n = 182) was used to validate the stunning procedure at the discovered anatomical site in the first phase, which was located 8 cm dorsal above the middle of the forehead on an "X" formed between the eyes and the base of the contralateral horns, and 2 cm lateral, avoiding the midline, where the skull tended to narrow. A total of 95.1% of buffaloes received effective stunning at the first shot with evidence of the presence of collapse, absence of rhythmic breathing, and absence of ocular reflexes (corneal and palpebral). There were no differences in the stunning efficacy by sex, breed, or skull thickness. These findings demonstrated that stunning with a PPCB at pressures of 1379-1516.8 KPa (200-220 pounds per square inch (psi)) in the site reported here produces a highly effective stunning at the first shot in water buffaloes.

299 Differences in Microbial Community Composition between Uterine Horns Ipsilateral and Contralateral to the Corpus Luteum in Beef Cows on day 15 of the Estrous Cycle

Abstract The objective of this study was to evaluate the uterine microbiota composition of cows on day 15 of the estrous cycle. Non-pregnant Bos taurus beef cows (n = 23) were exposed to a pre-synchronization step followed by the 7-d CO-Synch estrus synchronization protocol. On days -10, -3, 0 (exogenously induced ovulation), 7 and 14, transrectal ultrasonography was performed to evaluate ovarian structures, ensure synchrony, and determine the side of ovulation. Cows were harvested on day 15 and individual swabs were collected from each uterine horn using aseptic techniques. DNA was extracted and the entire (V1-V9 hypervariable regions) 16s rRNA gene was sequenced. Sequences were analyzed using the QIIME2 Pipeline. Across all samples, 22 phyla, 130 families, 215 genera, and 90 different species were identified. Butyrivibrio, Bacteroidales RF16 group, Clostridia_UCG-014, and Moraxellaceae, had different (P &amp;lt; 0.05) relative abundances in the ipsilateral compared with contralateral horns, whereas Proteobacteria, Acinetobacter, Bradyrhizobium, Lachnospiraceae FCS020 group, Nanoarchaeota, Woesearchaeales, and Prevotellaceae YAB2003 group tended (P &amp;lt; 0.10) to differ between ipsilateral and contralateral horns. In conclusion, there were significant differences in the composition of the microbial community of the ipsilateral and contralateral horn of cows on day 15 of the estrous cycle.

Surgical disorders in pediatric and adolescent gynecology: Vaginal and uterine anomalies.

Obstructive vaginal and uterine anomalies including imperforate hymen, transverse vaginal septum, and vaginal and/or cervical atresia or aplasia, might rarely present in infancy or childhood with hydrocolpos and/or hydrometra but they usually go unrecognized until presentation with amenorrhea and hematocolpos and/or hematometra in puberty. They should always be included in the differential diagnosis of a suprapubic and/or introital mass; in the latter case, vaginal vascular malformations and vaginal tumors should also be considered. Uterovaginal aplasia typically manifests with amenorrhea in puberty and needs to be differentiated from complete androgen insensitivity syndrome and gonadal dysgenesis of genetic males. Uterine fusion anomalies usually present with fertility and/or obstetrical complications in adulthood. However, a unicornuate uterus with a blind rudimentary contralateral horn containing functioning endometrium, and didelphys or septate uterus with a deviating obstructive septum might present in childhood or puberty with sequelae related with secretions or menstrual retention. This review provides a collective account of the most clinically important information about vaginal and uterine anomalies in childhood and adolescence for clinicians involved in the care of young females with the aim to provide guidance in appropriate evaluation and management.

Docosahexaenoic Acid and Melatonin Prevent Impaired Oligodendrogenesis Induced by Intrauterine Growth Restriction (IUGR).

In this study, our aims were to characterize oligodendrogenesis alterations in fetuses with intrauterine growth restriction (IUGR) and to find therapeutic strategies to prevent/treat them using a novel rabbit in vitro neurosphere culture. IUGR was surgically induced in one uterine horn of pregnant rabbits, while the contralateral horn served as a control. Neural progenitor cells (NPCs) were obtained from pup’s whole brain and cultured as neurospheres mimicking the basic processes of brain development including migration and cell differentiation. Five substances, chosen based on evidence provided in the literature, were screened in vitro in neurospheres from untreated rabbits: Docosahexaenoic acid (DHA), melatonin (MEL), zinc, 3,3′,5-Triiodo-L-thyronine (T3), and lactoferrin (LF) or its metabolite sialic acid (SA). DHA, MEL and LF were further selected for in vivo administration and subsequent evaluation in the Neurosphere Assay. In the IUGR culture, we observed a significantly reduced percentage of oligodendrocytes (OLs) which correlated with clinical findings indicating white matter injury in IUGR infants. We identified DHA and MEL as the most effective therapies. In all cases, our in vitro rabbit neurosphere assay predicted the outcome of the in vivo administration of the therapies and confirmed the reliability of the model, making it a powerful and consistent tool to select new neuroprotective therapies.

Unilateral uterine torsion in a preparturient Boer bUll bitch resolved by caesarian section and unilateral cornuectomy

Uterine torsion is a rare finding in dogs and is most often presented with a life-threatening condition. We hereby report a case of unilateral uterine torsion in a three-year old, pregnant bitch, two days to its expected date of delivery. The bitch became anorexic about two days prior to presentation and lumps of blood from vulva were found inside its kernel on the morning of presentation. The physiological parameters, except the rectal temperature (38.70C), were all outside the normal range for dogs. Haematology investigation revealed a packed cell volume (PCV) of 20% and hence, the need for haemaccel® administration. Digital evaluation of the birth canal with the middle finger only revealed a foetus with normal anterior presentation but no cervical dilation. Ultrasonography showed two live foetuses at the pelvic region of the bitch. Caesarean section was conducted according to standard technique to save the two puppies in the left contralateral horn. The right uterine horn was twisted along its longitudinal axis over 360o in a clockwise direction from inter-cornual junction. It had two dead foetuses and was filled with over 5 litres of clotted blood/dark fluid with a lot of necrotic tissue. The dead foetuses were removed and the right horn was excised after due consultation with the owner. The surgical incisions were closed according to standard techniques. Our timely intervention to do caesarean section and right unilateral cornuectomy saved the bitch and the two foetuses in the contralateral intact horn and partially preserved the reproductive potential of the bitch

Unilateral uterine torsion in a preparturient Boer bUll bitch resolved by caesarian section and unilateral cornuectomy

Uterine torsion is a rare finding in dogs and most often presented with a life-threatening condition. We hereby report a case of unilateral uterine torsion in a three-year old, pregnant bitch, two days to its expected date of delivery. The bitch became anorexic about two days prior to presentation and lumps of blood from vulva were found inside its kernel on the morning of the presentation. The physiological parameters, except the rectal temperature (38.70C), were all outside the normal range for dogs. Haematology investigation revealed a packed cell volume (PCV) of 20% and hence, the need for haemaccel® administration. Digital evaluation of the birth canal with the middle finger only revealed a foetus with the normal anterior presentation but no cervical dilation. Ultrasonography showed two live foetuses at the pelvic region of the bitch. Caesarean section was conducted according to standard technique to save the two puppies in the left contralateral horn. The right uterine horn was twisted along its longitudinal axis over 360o in a clockwise direction from the inter-cornual junction. It had two dead foetuses and was filled with over 5 litres of clotted blood/dark fluid with a lot of necrotic tissue. The dead foetuses were removed and the right horn was excised after due consultation with the owner. The surgical incisions were closed according to standard techniques. Our timely intervention to do caesarean section and right unilateral cornuectomy saved the bitch and the two foetuses in the contralateral intact horn and partially preserved the reproductive potential of the bitch

Unilateral uterine torsion in a preparturient Boer Bull bitch resolved by caesarian section and unilateral cornuectomy

Uterine torsion is a rare finding in dogs and most often presented with a life-threatening condition. We hereby report a case of unilateral uterine torsion in a three-year old, pregnant bitch, two days to its expected date of delivery. The bitch became anorexic about two days prior to presentation and lumps of blood from vulva were found inside its kernel on the morning of presentation. The physiological parameters, except the rectal temperature (38.70C), were all outside the normal range for dogs. Haematology investigation revealed a packed cell volume (PCV) of 20% and hence, the need for haemaccel® administration. Digital evaluation of the birth canal with the middle finger only revealed a foetus with normal anterior presentation but no cervical dilation. Ultrasonography showed two live foetuses at the pelvic region of the bitch. Caesarean section was conducted according to standard technique to save the two puppies in the left contralateral horn. The right uterine horn was twisted along its longitudinal axis over 360o in a clockwise direction from inter-cornual junction. It had two dead foetuses and was filled with over 5 litres of clotted blood/dark fluid with a lot of necrotic tissue. The dead foetuses were removed and the right horn was excised after due consultation with the owner. The surgical incisions were closed according to standard techniques. Our timely intervention to do caesarean section and right unilateral cornuectomy saved the bitch and the two foetuses in the contralateral intact horn and partially preserved the reproductive potential of the bitch.

Unilateral uterine torsion in a preparturient Boer Bull bitch resolved by caesarian section and unilateral cornuectomy

Uterine torsion is a rare finding in dogs and is most often presented with a life-threatening condition. We hereby report a case of unilateral uterine torsion in a three-year old, pregnant bitch, two days to its expected date of delivery. The bitch became anorexic about two days prior to presentation and lumps of blood from vulva were found inside its kernel on the morning of the presentation. The physiological parameters, except the rectal temperature (38.70C), were all outside the normal range for dogs. Haematology investigation revealed a packed cell volume (PCV) of 20% and hence, the need for haemaccel® administration. Digital evaluation of the birth canal with the middle finger only revealed a foetus with the normal anterior presentation but no cervical dilation. Ultrasonography showed two live foetuses at the pelvic region of the bitch. Caesarean section was conducted according to standard technique to save the two puppies in the left contralateral horn. The right uterine horn was twisted along its longitudinal axis over 360o in a clockwise direction from inter-cornual junction. It had two dead foetuses and was filled with over 5 litres of clotted blood/dark fluid with a lot of necrotic tissue. The dead foetuses were removed and the right horn was excised after due consultation with the owner. The surgical incisions were closed according to standard techniques. Our timely intervention to do caesarean section and right unilateral cornuectomy saved the bitch and the two foetuses in the contralateral intact horn and partially preserved the reproductive potential of the bitch.

Peroxiredoxin 6 Plays Essential Role in Mediating Fertilization and Early Embryonic Development in Rabbit Oviduct.

The oviduct is a site for early reproductive events including gamete maturation, fertilization, and early embryo development. Secretory cells lining the oviduct lumen synthesize and secrete proteins that interact with gametes and developing embryos. Although previous studies have identified some of the secretory proteins in the oviduct, however, knowledge and their precise specific functions in the oviduct are poorly understood. In this study, by using proteomic approach, we identified a secretory protein, Peroxiredoxin 6 (PRDX6), and evaluated its role in mediating early pregnancy events, fertilization, and embryo development in rabbit oviduct. The expression of PRDX6 was significantly higher in ampulla and isthmus sections of the oviduct in mated animal groups compared to non-mated controls. Furthermore, significant reduction in number of embryos recovered from PRDX6 siRNA-transfected oviductal horn was observed compared to the control contralateral horn. Moreover, in animals receiving PRDX6 siRNA in their oviductal horn, the number of implanted blastocysts was significantly less in the uterus as observed on day 9 post-coital (p.c.). Further, during embryo-rabbit oviduct epithelial cell (ROEC) co-culture, siRNA-mediated PRDX6 silencing attenuated the early embryonic development. Mechanistically, increased levels of ROS and expression of oxidative stress- and inflammation-related proteins were found in PRDX6 siRNA-treated ROEC cells as compared to control cells, implicating that ablation of PRDX6 in the oviduct creates a stress-induced micro-environment detrimental to early embryonic development in oviduct. Taken together, our data suggest that PRDX6 maintains an optimal micro-environment conducive to successful embryo development and can be considered as a candidate to evaluate its therapeutic potential in IVF strategies.

Long-Term Suppression of c-Jun and nNOS Preserves Ultrastructural Features of Lower Motor Neurons and Forelimb Function after Brachial Plexus Roots Avulsion.

Brachial plexus root avulsions cause debilitating upper limb paralysis. Short-term neuroprotective treatments have reported preservation of motor neurons and function in model animals while reports of long-term benefits of such treatments are scarce, especially the morphological sequelae. This morphological study investigated the long-term suppression of c-Jun- and neuronal nitric oxide synthase (nNOS) (neuroprotective treatments for one month) on the motor neuron survival, ultrastructural features of lower motor neurons, and forelimb function at six months after brachial plexus roots avulsion. Neuroprotective treatments reduced oxidative stress and preserved ventral horn motor neurons at the end of the 28-day treatment period relative to vehicle treated ones. Motor neuron sparing was associated with suppression of c-Jun, nNOS, and pro-apoptotic proteins Bim and caspases at this time point. Following 6 months of survival, neutral red staining revealed a significant loss of most of the motor neurons and ventral horn atrophy in the avulsed C6, 7, and 8 cervical segments among the vehicle-treated rats (n = 4). However, rats that received neuroprotective treatments c-Jun JNK inhibitor, SP600125 (n = 4) and a selective inhibitor of nNOS, 7-nitroindazole (n = 4), retained over half of their motor neurons in the ipsilateral avulsed side compared. Myelinated axons in the avulsed ventral horns of vehicle-treated rats were smaller but numerous compared to the intact contralateral ventral horns or neuroprotective-treated groups. In the neuroprotective treatment groups, there was the preservation of myelin thickness around large-caliber axons. Ultrastructural evaluation also confirmed the preservation of organelles including mitochondria and synapses in the two groups that received neuroprotective treatments compared with vehicle controls. Also, forelimb functional evaluation demonstrated that neuroprotective treatments improved functional abilities in the rats. In conclusion, neuroprotective treatments aimed at suppressing degenerative c-Jun and nNOS attenuated apoptosis, provided long-term preservation of motor neurons, their organelles, ventral horn size, and forelimb function.

Structural Brain Changes during the Neonatal Period in a Rabbit Model of Intrauterine Growth Restriction

Background: Intrauterine growth restriction (IUGR) is associated with abnormal neurodevelopment, but the associated structural brain changes are poorly documented. The aim of this study was to describe in an animal model the brain changes at the cellular level in the gray and white matter induced by IUGR during the neonatal period. Methods: The IUGR model was surgically induced in pregnant rabbits by ligating 40–50% of the uteroplacental vessels in 1 horn, whereas the uteroplacental vessels of the contralateral horn were not ligated. After 5 days, IUGR animals from the ligated horn and controls from the nonligated were delivered. On the day of delivery, perinatal data and placentas were collected. On postnatal day 1, functional changes were first evaluated, and thereafter, neuronal arborization in the frontal cortex and density of pre-oligodendrocytes, astrocytes, and microglia in the corpus callosum were evaluated. Results: Higher stillbirth in IUGR fetuses together with a reduced birth weight as compared to controls was evidenced. IUGR animals showed poorer functional results, an altered neuronal arborization pattern, and a decrease in the pre-oligodendrocytes, with no differences in microglia and astrocyte densities. Conclusions: Overall, in the rabbit model used, IUGR is related to functional and brain changes evidenced already at birth, including changes in the neuronal arborization and abnormal oligodendrocyte maturation.

4 Aryl hydrocarbon receptor targets are upregulated in porcine blastocyst-stage embryos that were cultured invitro: A transcriptional analysis

Invitro-produced (IVP) porcine embryos are developmentally delayed compared with those derived invivo. Thus, efforts have been made to modify the current medium for IVP of porcine embryos in an attempt to shift the abundance of target transcripts towards the invivo level. The objective of the current study was to identify differences in mRNA abundance that may account for reduced developmental competence in IVP embryos and to determine whether alterations to maturation and culture media directed the transcriptional profiles of IVP embryos towards an invivo state. Following AI of gilts, an oviduct and tip of the uterine horn were flushed on Day 2 to recover 4-cell stage embryos; these were cultured for 4 days in MU3, generating IVM and invitro-cultured (IVC) blastocyst-stage embryos. On Day 6, the gilts were killed, and the contralateral horns were flushed to obtain invivo-derived (IVV) blastocyst-stage embryos. The third group of blastocyst-stage embryos, referred to as invitro-matured and cultured (IVMC), were created by aspirating cumulus–oocyte complexes from slaughterhouse-derived ovaries, maturing and fertilizing invitro, and culturing for 6 days in MU3. Total RNA was extracted from pools of 10 blastocyst-stage embryos with 3 replicates per group. First- and second-strand cDNA was synthesised and sequenced by using the Illumina platform (Illumina Inc.). After removal of adapters and reads that mapped to porcine rRNA genes and the PhiX genome, reads were mapped to the Sus scrofa genome by using STAR (version 2.7.1a) with default options. Pairwise comparisons were performed to test for differential expression of genes by using the Bioconductor package DESEqn 2. Transcripts were differentially abundant (false discovery rate &amp;lt;0.05) between IVV and IVC embryos (2,450), between IVV and IVMC embryos (3,045), and between IVC and IVMC embryos (262). Pathways related to cell cycle were downregulated in IVC and IVMC compared with IVV embryos, and pathways related to amino acid transport in metabolism were upregulated in IVC and IVMC compared with IVV embryos. Of particular interest, message for cytochrome P450 family 1 subfamily A member 1 (CYP1A1) was only present in the IVC (956 reads) and IVMC (381 reads) embryos but not in the IVV embryos (0 reads). The aryl hydrocarbon receptor (AHR) promotes transcription of CYP1A1, which encodes a monooxygenase involved in xenobiotic metabolism. Moreover, abundance of 12 other AHR targets was increased in IVC and IVMC embryos (false discovery rate &amp;lt;0.05) compared with IVV embryos. Thus, production of porcine embryos invitro may activate AHR, resulting in altered transcriptional profiles and reduced competence. This research was funded by USDA-NIFA (2019-67011-29543).

Sperm transport and endometrial inflammatory response in mares after artificial insemination with cryopreserved spermatozoa

This study aimed to determine whether the insemination site and dose with cryopreserved sperm of reproductively normal mares affect the sperm population in uterine tubes and the intensity of endometrial inflammatory response. Experimental subjects were estrous mares inseminated, in the mid-uterine body (Body) or the tip of the uterine horn (Tip), ipsilateral to the dominant follicle, with one 0.5 mL straw with 50 × 106 sperm (50) or with eight straws with 50 × 106 sperm/straw (400). Mares were slaughtered 2 h, 4 h and 12 h after artificial insemination (AI) and randomly assigned to following groups: Body 50 (n = 19) (2 h, 4 h or 12 h); Tip 50 (n = 29) (2 h, 4 h, or 12 h); Body 400 (n = 24) (2 h, 4 h, or 12 h); Tip 400 (n = 21) (2 h, 4 h, or 12 h). A Control group (n = 16) was not inseminated. After slaughter, uterine tubes were separated from uterus, and uteri and tubes flushed with phosphate-buffered saline (PBS). After flushing, an endometrial sample was collected from ipsilateral and contralateral horns and mid-uterus body for further histopathological examination. A sample of each uterine tube flushing was examined for sperm count, and a sample of each uterine flushing was used for polymorphonuclear neutrophils (PMNs) count. Data were analyzed using PROC GLM from SASv9.4. Insemination time, site, sperm dose, and their interactions were considered independent variables and sperm and PMNs numbers dependent variables. Deep horn insemination increased ipsilateral uterine tube sperm number without an increase in the inflammatory reaction compared with the uterine body insemination. The higher the insemination dose, the higher the uterine tubes’ sperm number and inflammatory reaction, with a quicker resolution. In conclusion, the insemination site and dose affected sperm in the uterine tubes, while post-insemination time and dose influenced the inflammatory reaction.