Uterine Protein Research Articles

Cell-Free Fetal DNA, Telomeres, and the Spontaneous Onset of Parturition.

Multiple previous reports have provided compelling support for the premise that spontaneous parturition is mediated by activation of inflammation-related signaling pathways leading to increased secretion of cytokines and chemokines, the influx of neutrophils and macrophages into the pregnant uterus, increased production of uterine activation proteins (eg, connexin-43, cyclo-oxygenase-2, oxytocin receptors, etc), activation of matrix metalloproteinases, and the release of uterotonins leading to cervical ripening, membrane rupture, and myometrial contractions. The missing link has been the fetal/placental signal that triggers these proinflammatory events in the absence of microbial invasion and intrauterine infection. This article reviews the biomedical literature regarding the increase in cell-free fetal DNA (cffDNA), which is released during apoptosis in the placenta and fetal membranes at term, the ability of apoptosis modified vertebrate DNA to stimulate toll-like receptor-9 (TLR9) leading to increased release of cytokines and chemokines, and the potential "fail-safe" role for the anti-inflammatory cytokine IL-10. This article also reviews the literature supporting the key role that telomere loss plays in regard to increasing the ability of vertebrate (including placental) DNA to stimulate TLR9, and in regard to signaling the onset of apoptosis in the placenta and fetal membranes, thereby providing a biologic clock that determines the length of gestation and the timing for the onset of parturition. In summary, this literature review provides a strong rationale for future research to test the hypothesis that telomere loss and increased cffDNA levels trigger the proinflammatory events leading to the spontaneous onset of parturition in mammals: the "cffDNA/telomere hypothesis."

Novel Noncompetitive IL-1 Receptor-Biased Ligand Prevents Infection- and Inflammation-Induced Preterm Birth.

Preterm birth (PTB) is firmly linked to inflammation regardless of the presence of infection. Proinflammatory cytokines, including IL-1β, are produced in gestational tissues and can locally upregulate uterine activation proteins. Premature activation of the uterus by inflammation may lead to PTB, and IL-1 has been identified as a key inducer of this condition. However, all currently available IL-1 inhibitors are large molecules that exhibit competitive antagonism properties by inhibiting all IL-1R signaling, including transcription factor NF-κB, which conveys important physiological roles. We hereby demonstrate the efficacy of a small noncompetitive (all-d peptide) IL-1R-biased ligand, termed rytvela (labeled 101.10) in delaying IL-1β-, TLR2-, and TLR4-induced PTB in mice. The 101.10 acts without significant inhibition of NF-κB, and instead selectively inhibits IL-1R downstream stress-associated protein kinases/transcription factor c-jun and Rho GTPase/Rho-associated coiled-coil-containing protein kinase signaling pathways. The 101.10 is effective at decreasing proinflammatory and/or prolabor genes in myometrium tissue and circulating leukocytes in all PTB models independently of NF-κB, undermining NF-κB role in preterm labor. In this work, biased signaling modulation of IL-1R by 101.10 uncovers a novel strategy to prevent PTB without inhibiting NF-κB.

Uterine expression of tight junctions in the Canine uterus

Tight junctions (TJs) form continuous intercellular contacts in intercellular junctions. TJs involve integral proteins such as occludin (OCLN) and claudins (CLDNs) as well as peripheral proteins such as zona occludens-1 (ZO-1) and junctional adhesion molecules (JAMs). TJs control paracellular transportation across cell-to-cell junctions. Although TJs have been studied for several decades, comparison of the transcriptional-translational levels of these molecules in canine organs has not yet been performed. In this study, we examined uterine expression of CLDNs, OCLN, junction adhesion molecule-A, and ZO-1 in canine. Expression levels of canine uterine TJ proteins, including CLDN1, 2, 4, 5, JAM-A, ZO-1, and OCLN, were measured using reverse transcription PCR, real-time PCR, and Western blotting, whereas TJs distribution was determined by immunohistochemistry. The mRNA and protein expression levels of OCLN, CLDN-1, 4, JAM-1, and ZO-1 were identified in the uterus. Immunohistochemistry demonstrated that TJs were localized to the endometrium and/or myometrium of the uterus. Our results show that canine TJ proteins, including CLDNs, OCLN, JAM-A, and ZO-1, were expressed in the canine uterus. Taken together, these proteins may perform unique physiological roles in the uterus. Therefore, these findings may serve as a basis for further studies on TJ proteins and their roles in the physiological or pathological condition of the canine uterus.

Deficiency of monoclonal non-specific suppressor factor beta (MNSFB) promotes pregnancy loss in mice.

Maternal immune tolerance to the semi-allogenic fetus is required for successful pregnancy in mammals. Monoclonal nonspecific suppressor factor beta (MNSFB) is an immunosuppressive factor present in uterine epithelial and stromal cells, as well as in macrophages and T cells. Although the functional neutralization of MNSFB using specific antibodies against it lead to failed embryo implantation in mice, the exact role of MNSFB at the fetal-maternal interface remains unclear. The present study generated conditional heterozygous Mnsfb-deficient (Mnsfb(+/) (-) ) mice using the LoxP/Cre system. Western-blot analyses showed that uterine MNSFB protein in Mnsfb(+/-) mice was remarkably down-regulated compared to that in the wild-type (Mnsfb(+/+) ) mice. The litter size of female Mnsfb(+/-) mice was significantly reduced, which corresponded to developmental failure of embryos beyond Day 11 of pregnancy. The expression level of MNSFB protein was also lower in the failing compared to the normal embryos. An aberrant interaction between the embryos of Day-4 pregnant wild-type mice and endometrial stromal cells of female Mnsfb(+/-) mice was observed in vitro. The uterine Day-5 abundance of P53, BAX, and BCL-G in pregnant Mnsfb(+/-) mice was significantly decreased compared to that of wild-type mice, whereas the expression of P27 and tumor necrosis factor alpha (TNFA) was elevated. By comparison, the levels of MNSFB and BAX proteins in human decidual tissues obtained from recurrent spontaneous miscarriage patients were significantly reduced compared to those obtained from legal medial abortion, highlighting the involvement of MNSFB in the pathogenesis of recurrent spontaneous miscarriage. Together, these results demonstrated that a deficiency in MNSFb is associated with pregnancy loss, probably through reduced P53 and/or increased TNFA production at the fetal-maternal interface.

Comparative effects of the crude methanol/methylene chloride extract and fractions of Senecio biafrae (Oliv. & Hiern) J. Moore on some fertility parameters in immature female Wistar rats

ObjectiveTo assay the comparative effect of Senecio biafrae leaves and stems methanol/methylene chloride extract as well as its hexane, n-butanol and ethyl acetate fractions on reproductive function of immature female rat. MethodsVarious doses of the methanol/methylene chloride extract and fractions were orally administered for 20 consecutive days to immature female rats. The parameters of the rats such as body, ovarian and uterus weight, uterine, ovarian and serum proteins, ovarian cholesterol as well as follicle-stimulating hormone, luteinizing hormone, estradiol and progesterone levels were recorded. ResultsLevels of ovarian cholesterol and protein as well as uterine protein were doubled principally in animals treated with the ethyl acetate fraction not matter what dose was administered. The animals treated with crude extract presented a significant increase (P < 0.05) of 108%, 79% and 135% in serum estradiol level at the dose of 8 mg/kg, 32 mg/kg and 64 mg/kg respectively. ConclusionsResults of the present study provide evidence on the inducing effect of the methanol/methylene chloride extract of S. biafrae and its fractions on immature female rat's fertility parameters in serum, ovary and uterus.

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Objectives Studies have demonstrated abnormal pulse wave analysis (PWA) at the time of and after the diagnosis of pre-eclampsia. However, there remains a paucity of data concerning its predictive value. In addition, new cuff-based technology in measuring PWA is now available, removing the need for using applanation tonometry, which is highly operator-dependent. The aim of our study was to assess the utility of first trimester PWA using a cuff-based device and its potential role in predicting early-onset pre-eclampsia (delivery before 34 weeks). Methods Early-onset pre-eclampsia risk is currently determined by an algorithm consisting of maternal history, mean arterial pressure, uterine artery pulsatility index and pregnancy-associated plasma protein A at 11-13+6 weeks gestation. PWA was measured using a cuff-based device (SphygmoCor XCEL) at this time point. The differences in augmentation pressure (AP), augmentation index (AIx), and the heart-rate corrected augmentation index (AIx75) amongst the different risk groups were determined using t -tests. Results 746 women were studied. The mean age was 33.0 years (95% CI: 32.6 – 33.3 years). 132 were found to have ⩾1% risk (group A), and 36 had ⩾5% risk (group B). For group A, all three parameters were elevated but not statistically significant compared to women with p = 0.08), AIx: 15.2 vs. 12.8 ( p = 0.13), AIx75: 19.8 vs. 17.2 ( p = 0.11)]. For group B, all three parameters were significantly elevated compared to women with p = 0.006), AIx: 19.1 vs. 12.9 ( p = 0.02), AIx75: 23.9 vs. 17.4 ( p = 0.02)]. Conclusions Women found to be at high risk of developing early pre-eclampsia were also found to have higher PWA parameters. This suggests that the generalized vascular changes associated with pre-eclampsia are present at this early stage of pregnancy. This simple technique may be useful in assessing the risk of developing early-onset pre-eclampsia. Disclosures P. Lan: None. J. Hyett: None. A. Gillin: None.

Cloprostenol and pregnant mare serum gonadotropin promote estrus synchronization, uterine development, and follicle-stimulating hormone receptor expression in mice.

This study investigated the effects of pregnant mare se-rum gonadotropin (PMSG) and cloprostenol (CLO) on estrus induc-tion and synchronization, uterine development, and follicle-stimulating hormone receptor (FSHR) expression in mice. A total of 105 Kunming pre-puberty mice were divided into seven subgroups. Three PMSG sub-groups were injected intraperitoneally with 10, 20, and 40 IU PMSG twice (on days 0 and 4), and three CLO subgroups were injected intra-peritoneally with 10, 15, and 20 μg cloprostenol acetate twice (on days 0 and 4). The results showed that 93.33 and 66.67% of synchronized mice displayed estrus within 18.68-37.59 h following CLO and PMSG exposure, respectively. Estrus numbers, estrus onset time, and estrus rates in CLO and PMSG groups were greater than in control groups (CG) (P < 0.05). Uterine weights of the PMSG group were higher than that of CLO and CG groups, and the uterine horn longitudinal diameters in experimental mice were greater than CG. Expression levels of FSHR proteins in CLO and PMSG groups increased slightly when compared to CG. In conclusion, CLO and PMSG administration did not clearly af-fect the expression of uterine FSHR proteins in mice. Moreover, PMSG and CLO treatments synchronized estrus and enhanced the uterine de-velopment of mice. The efficacy of CLO on estrus synchronization was greater than PMSG, and the effects of PMSG on uterine development were stronger than CLO. These results have important significance re-garding the modulation of animal reproductive functions.

Effect of the aqueous extract of Senecio biafrae (Oliv. & Hiern) J. Moore on some fertility parameters in immature female rat

Ethnopharmacological relevanceSenecio biafrae is a plant from the huge family of Asteraceae used in the African pharmacopoeia for the treatment of many ailments among which is infertility. Material and methodsThe aqueous extract, which was primarily subjected to polyphenol analysis, has been administered to immature female rats for 20 days at 8, 32, 64 and 128mg/kg of body weight. The day following the treatment, the animals were sacrificed; their serum, ovary and uterus were retained respectively for reproductive hormones, ovarian and uterine proteins, and ovarian cholesterol assays. ResultsLight body weight gain variation of treated animals was observed during the experimental period. A significant increase (p˂0.05) in serum estradiol and proteins as well as in uterine weight (p˂0.01) of all Senecio biafrae treated animals was noted. No significant variation was noticed in the ovarian weight and follicle numbers. ConclusionThe various biochemical and physiological parameters of fertility were significantly improved with the aqueous extract of Senecio biafrae, thus attesting some of its traditional usage.

Persistent Müllerian duct syndrome (PMDS) in the Polish free-ranged bull populations of the European bison (Bison bonasus L.)

This study describes the diversity of vestigial male uteri of the European bison (Eb) examined for: (1) morphology, (2) glycoprotein localization, (3) total protein and glycoprotein profiles, (4) steroid concentrations, and (5) PMDS based on the mutation of AMH and AMHR2 genes. Uteri of adult bulls (5–12 years old) were compared to a uterus of a juvenile female (6 months old). Male uterine proteins were analyzed in parallel to secretory endometrial proteins of pseudo-pregnant pig (PsEND) and BSA used as profile-controls. Hematoxylin/eosin-staining revealed the diversity of male uterine morphology, including lumen size/shape, endometrial (END) gland density, luminal knob-like epithelial structures and multiple intrauterine cells proliferating within the lumen. PAS-staining revealed the presence of glycoproteins restricted to luminal epithelial cells and END glands. Heterologous total protein PAGE-profiles (20–66kDa) revealed two dominant fractions (66 and 45kDa), similar to PAS-profiles (67 and 47kDa) in male and female uterine tissues. In male uterine tissues, androstendione and progesterone, but not testosterone, estrone or estradiol concentrations were lower than in the female. Sequencing of AMH- and AMHR2-like amplicons allowed identification of these gene mutations in Eb. Our results provide novel data regarding PMDS, demonstrating the diversity of uterine morphology, glycoprotein mass/profile, steroid concentration and AMH/AMHR2 mutations in Eb bulls.

Prostaglandin F2α regulates the expression of uterine activation proteins via multiple signalling pathways.

Prostaglandin F2α (PGF2A) has multiple roles in the birth process in addition to its vital contractile role. Our previous study has demonstrated that PGF2A can modulate uterine activation proteins (UAPs) in cultured pregnant human myometrial smooth muscle cells (HMSMCs). The objective of this study was to define the signalling pathways responsible for PGF2A modulation of UAPs in myometrium. It was found that PGF2A stimulated the expression of (GJA1) connexin 43 (CX43), prostaglandin endoperoxide synthase 2 (PTGS2) and oxytocin receptor (OTR) in cultured HMSMCs. The inhibitors of phospholipase C (PLC) and protein kinase C (PKC) blocked PGF2A-stimulated expression of CX43. The inhibitors of ERK, P38 and NFκB also blocked the effect of PGF2A on CX43 expression, whereas PI3K and calcineurin/nuclear factor of activated T-cells (NFAT) pathway inhibitors did not reverse the effect of PGF2A on CX43. For PTGS2 and OTR, PLC, PI3K, P38 and calcineurin/NFAT signalling pathways were involved in PGF2A action, whereas PKC and NFκB signalling were not involved. In addition, PGF2A activated NFAT, PI3K, NFκB, ERK and P38 signalling pathways. Our data suggest that PGF2A stimulates CX43, PTGS2 and OTR through divergent signalling pathways.

Data set for the proteomic inventory and quantitative analysis of chicken uterine fluid during eggshell biomineralization.

Chicken eggshell is the protective barrier of the egg. It is a biomineral composed of 95% calcium carbonate on calcitic form and 3.5% organic matrix proteins. Mineralization process occurs in uterus into the uterine fluid. This acellular fluid contains ions and organic matrix proteins precursors which are interacting with the mineral phase and control crystal growth, eggshell structure and mechanical properties. We performed a proteomic approach and identified 308 uterine fluid proteins. Gene Ontology terms enrichments were determined to investigate their potential functions. Mass spectrometry analyses were also combined to label free quantitative analysis to determine the relative abundance of 96 proteins at initiation, rapid growth phase and termination of shell calcification. Sixty four showed differential abundance according to the mineralization stage. Their potential functions have been annotated. The complete proteomic, bioinformatic and functional analyses are reported in Marie et al., J. Proteomics (2015) [1].

Quantitative proteomics and bioinformatic analysis provide new insight into protein function during avian eggshell biomineralization.

The calcitic avian eggshell protects the developing embryo and, moreover, ensures that the nutritious table egg remains free of pathogens. The eggshell is formed by nucleation upon a fibrous scaffold (the eggshell membranes) followed by an interaction between the growing mineral crystals and the shell organic matrix. This interaction leads to a highly ordered shell microstructure and texture which contribute to its exceptional mechanical properties. Shell mineralization occurs in three distinct phases of calcification (initiation, growth and termination), which are associated with distinct populations of matrix proteins that are secreted into the acellular uterine fluid as modulators of the process. The recent development of high-throughput methods has led to the identification of many proteins in the shell, but little is known concerning their role in shell formation. In order to determine precisely the importance of particular proteins relative to eggshell mineralization, this project used qualitative and quantitative proteomics of the uterine fluid constituents, coupled with bioinformatic analysis, to predict the functional role of proteins secreted at each of the three main stages of shell calcification. Besides its relevance to food production and to hen reproduction, eggshell calcification is furthermore a relevant model for studying calcium carbonate biomineralization on a two-dimensional membrane support. Better understanding of this process will provide insight into the fabrication of ceramics at ambient pressure and temperature.

Effects of GnRHa active immunity on FSHR expression and uterine development in prepuberty and non cyclic ewes

Summary This study explored the effects of GnRH agonist (GnRHa) immunity on expression of FSHR mRNA in pituitary and FSHR protein in uteri, and also confirmed its efficacy on uterine development. 42 prepuberty and non cyclic ewes were assigned to 6 experimental groups (EG) during anestrus season. Animals in EG-I, EG-II and EG-III were subcutaneously injected with 200, 300 and 400 µg alarelin antigens (day 0 and 14). Animals in EG-IV and EG-V were subcutaneously injected with 200 and 300 µg alarelin antigen (day 0, 7, 14 and 21). Uterine FSHR protein expression was detected using Western-blotting. RT-PCR was implemented to measure expression of pituitary FSHR mRNA on day 70 after the immunity. Compared to the CG, pituitary FSHR mRNA levels reduced by 58%, 88% and 91% (P<0.01) in EG-I, EG-II and EG-III. FSHR mRNA in EG-IV and EG-V reduced by 64.29% (P<0.05) and 10.11% compared to EG-I and EG-II. Expression levels of uterine FSHR protein in EG-III and EG-V were higher than that in CG. The uterine glands, mitochondria and mitochondrial crista decreased, and the glandular cavity reduced, also the microvilli shortened. Uterine horn weights in EG-I, EG-II and EG-III reduced by 4.66%, 10.20% and 16.63%, respectively. Uterine wall thickness (UWT) in EG-I, EG-II and EG-III reduced by 1.41%, 5.84% and 8.75% (P<0.05). The endometrial epithelium thickness (EET) in EG-I, EG-II and EG-III decreased 2.70%, 8.87% and 19.15%. The positive correlations between uterine horn weights and UWT, EET and FSHR were calculated.

Reproductive seasonality in the Indian pygmy field mouse, Mus terricolor

The Indian pygmy field mouse, Mus terricolor, is a tiny, yet economically and ecologically important crop pest found throughout South-East Asia. There are no systematic reports exploring its reproductive physiology. We report the presence of distinct periods of annual reproductive activity and quiescence in M. terricolor. Body weight in males and females, relative weights of testis, epididymis and seminal vesicle in males, ovarian and uterine weight in females, gonadal histomorphic changes, testicular and ovarian cholesterol, sialic acid in epididymis, fructose in seminal vesicle, uterine protein content, melatonin in males and females, testosterone in males, estradiol, and progesterone in females were studied over a period of three years in both wild-caught and lab-acclimated mice. The number of Graafian follicles and corpora lutea, and plasma estradiol and progesterone, along with relative weights of ovary and uterus in females exhibited a peak in the months of October–January, compared to June. Based on histomorphic and hormonal status, the major reproductively active season is the winter (short-day breeding). There is a brief period of sub-maximal reproductive activity in April. M. terricolor is reproductively inactive in the summer, monsoons, and autumn. The results establish M. terricolor as a seasonal breeder in the field, with interesting implications for pest management.

Altered gene expression patterns during the initiation and promotion stages of neonatally diethylstilbestrol-induced hyperplasia/dysplasia/neoplasia in the hamster uterus.

Neonatal treatment of hamsters with diethylstilbestrol (DES) induces uterine hyperplasia/dysplasia/neoplasia (endometrial adenocarcinoma) in adult animals. We subsequently determined that the neonatal DES exposure event directly and permanently disrupts the developing hamster uterus (initiation stage) so that it responds abnormally when it is stimulated with estrogen in adulthood (promotion stage). To identify candidate molecular elements involved in progression of the disruption/neoplastic process, we performed: (1) immunoblot analyses and (2) microarray profiling (Affymetrix Gene Chip System) on sets of uterine protein and RNA extracts, respectively, and (3) immunohistochemical analysis on uterine sections; all from both initiation stage and promotion stage groups of animals. Here we report that: (1) progression of the neonatal DES-induced hyperplasia/dysplasia/neoplasia phenomenon in the hamster uterus involves a wide spectrum of specific gene expression alterations and (2) the gene products involved and their manner of altered expression differ dramatically during the initiation vs. promotion stages of the phenomenon.

Morphofunctional alterations of the nonpregnant murine uterus in response to intense and exhaustive exercise are not related to oxidative stress

Exercise is a common and noninvasive way to improve human health. In contrast, intense exercise causes damage in various tissues and is usually associated with metabolic changes in organs and tissues. Even though intense exercise is associated with dysfunctions in the female reproductive system, much less is known about the cellular mechanisms underlying its effects particularly on the nonpregnant uterus. We investigated whether the effects of an intense and exhaustive exercise (IEE) program on the isolated C57BL/6 uterine morphology and contractility might be related to increased levels of prooxidation markers. Female mice were submitted to 2 days of IEE. The daily exercise session consisted of a running session until exhaustion, with the treadmill speed set at 85% of each animal's maximum velocity. Training responses were evaluated through two parameters: time to exhaustion and maximum velocity. Absence of exercise-induced hypothalamic-pituitary-adrenal (HPA) axis activation was indirectly evaluated by maintenance of the adrenal gland weight. IEE reduced the thickness of the longitudinal muscular layer by 10%, impaired contractility in response to muscarinic stimulation (increased EC50 and decreased Emax), but showed a strong trend to decreasing the KCl-induced contraction; reduced lipid peroxidation; and did not alter the uterine protein oxidation of exercised animals compared with control. Altogether we provide evidence for the nonpregnant murine uterus being an important target to IEE, leading to morphofunctional alterations which could not be associated with tissue oxidative stress but might well be related with exercise-induced uterine dysfunctions.

Melatonin prevents experimental preterm labor and increases offspring survival

Preterm delivery is the leading cause of neonatal mortality and contributes to delayed physical and cognitive development in children. At present, there is no efficient therapy to prevent preterm labor. A large body of evidence suggests that intra-amniotic infections may be a significant and potentially preventable cause of preterm birth. This work assessed the effect of melatonin in a murine model of inflammation-associated preterm delivery which mimics central features of preterm infection in humans. For this purpose, preterm labor was induced in BALB/c mice by intraperitoneal injections of bacterial lipopolysaccharide (LPS) at 10.00 hr (10 μg LPS) and 13.00 hr (20 μg LPS) on day 15 of pregnancy. On day 14 of pregnancy, a pellet of melatonin (25 mg) had been subcutaneously implanted into a group of animals. In the absence of melatonin, a 100% incidence of preterm birth was observed in LPS-treated animals, and the fetuses showed widespread damage. By comparison, treatment with melatonin prevented preterm birth in 50% of the cases, and all pups from melatonin-treated females were born alive and their body weight did not differ from control animals. Melatonin significantly prevented the LPS-induced rises in uterine prostaglandin (PG) E2 , PGF2α, and cyclooxygenase-2 protein levels. In addition, melatonin prevented the LPS-induced increase in uterine nitric oxide (NO) production, inducible NO synthase protein, and tumor necrosis factor-alpha (TNFα) levels. Collectively, our results suggest that melatonin could be a new therapeutic tool to prevent preterm labor and to increase offspring survival.

Effect of Exogenous Gonadal Steroids on Reproductive Functions of the Indian Pygmy Field Mouse Mus terricolor

Different thresholds of gonadal steroids exert stimulatory or inhibitory effects on GnRH and gonadotropin release. During the reproductively active phase (RAP), the concentration of endogenous gonadal steroids remains high while, during reproductively inactive phase (RIP), it remains low. During RIP the HPG axis is sensitive to gonadal steroids but the circulatory levels of testosterone and estradiol remain low. During this phase one can observe conveniently the effects of gonadal steroids on the HPG axis in male or female rodents. Therefore, the aim of the present study was to find the effects of testosterone and estradiol on the reproductive functions of male and female Indian pygmy field mouse, Mus terricolor , during RIP. The male mice were injected aquaviron (commercially available testosterone, 1mg/100g body weight) while the female mice received estradiol-benzoate (25μg/100g body weight) for 15 consecutive days during the RIP. After completion of the treatment, a significant increase in the weights of gonads and accessory sex organs was noted in both the sexes. The biochemical constituents of accessory sex organs such as epididymal sialic acid, seminal vesicular fructose and uterine protein content reflected significant elevation accompanied by increased levels of plasma testosterone, estradiol and progesterone. Histologically, the gonads and accessory sex organs exhibited increased cellular activity. However, the gonadal cholesterol was significantly decreased in both the sexes. Over all, administration of gonadal steroids to both male and female mice accelerated the gonadal recrudescence but did not inhibit the reproductive functions when administered during the RIP. Therefore, it can be inferred from the present study that during the RIP the HPG axis is sensitive to gonadal steroids and, hence, exogenous gonadal steroids induce gonadal activity.

Local Effects of Pregnancy on Connexin Proteins That Mediate Ca 2+ -Associated Uterine Endothelial NO Synthesis

Uterine artery adaptations during gestation facilitate increases in uterine blood flow and fetal growth. local expression and distribution of uterine artery connexins play roles in mediating in vivo gestational eNOS activation and NO production. We established an ovine model restricting pregnancy to a single uterine horn and measured uterine blood flow, uterine artery shear stress, connexins 37/43, and P(635)eNOS protein levels in uterine artery and systemic artery (omental and renal) endothelium and connexins in vascular smooth muscle. Uterine blood flow and shear stress were locally (unilaterally) and substantially elevated by gestation. During pregnancy, uterine artery endothelial gap junction proteins connexins 37/43 were locally regulated in the gravid horn and elevated 10.3- and 25.6-fold; uterine artery endothelial P(635)eNOS and total eNOS were elevated 3.3- and 2.9-fold; whereas uterine artery vascular smooth muscle connexins 37/43 were locally elevated 12.5- and 5.9-fold, respectively. Less pronounced changes were observed in systemic vasculature except for significant pregnancy-associated increases in omental artery vascular smooth muscle connexin 43 and omental artery endothelial P(635)eNOS and total eNOS. Gap junction blockade using connexin 43, but not connexin 37-specific Gap peptides, abrogated uterine artery endothelial ATP-induced Ca(2+)-mediated NO production. Thus, uterine artery endothelial connexin 43, but not connexin 37, regulates Ca(2+)-mediated NO production required for the vasodilation to accommodate increases in uterine blood flow and shear stress during healthy pregnancies.

Differential label‐free quantitative proteomic analysis of avian eggshell matrix and uterine fluid proteins associated with eggshell mechanical property

Eggshell strength is a crucial economic trait for table egg production. During the process of eggshell formation, uncalcified eggs are bathed in uterine fluid that plays regulatory roles in eggshell calcification. In this study, a label-free MS-based protein quantification technology was used to detect differences in protein abundance between eggshell matrix from strong and weak eggs (shell matrix protein from strong eggshells and shell matrix protein from weak eggshells) and between the corresponding uterine fluids bathing strong and weak eggs (uterine fluid bathing strong eggs and uterine fluid bathing weak eggs) in a chicken population. Here, we reported the first global proteomic analysis of uterine fluid. A total of 577 and 466 proteins were identified in uterine fluid and eggshell matrix, respectively. Of 447 identified proteins in uterine fluid bathing strong eggs, up to 357 (80%) proteins were in common with proteins in uterine fluid bathing weak eggs. Similarly, up to 83% (328/396) of the proteins in shell matrix protein from strong eggshells were in common with the proteins in shell matrix protein from weak eggshells. The large amount of common proteins indicated that the difference in protein abundance should play essential roles in influencing eggshell strength. Ultimately, 15 proteins mainly relating to eggshell matrix specific proteins, calcium binding and transportation, protein folding and sorting, bone development or diseases, and thyroid hormone activity were considered to have closer association with the formation of strong eggshell.