Non-pregnant Rat Uterus Research Articles

Three-dimensional virtual histology of the rat uterus musculature using micro-computed tomography.

Contractions of the uterus play an important role in menstruation and fertility, and contractile dysfunction can lead to chronic diseases such as endometriosis. However, the structure and function of the uterus are difficult to interrogate in humans, and thus animal studies are often employed to understand its function. In rats, anatomical studies of the uterus have typically been based on histological assessment, have been limited to small segments of the uterine structure, and have been time-consuming to reconstruct at the organ scale. This study used micro-computed tomography imaging to visualise the muscle structures in the entire non-pregnant rat uterus and assess its use for 3D virtual histology. An assessment of the rodent uterus is presented to (i) quantify muscle thickness variations along the horns, (ii) identify predominant fibre orientations of the muscles and (iii) demonstrate how the anatomy of the uterus can be mapped to 3D volumetric meshes via virtual histology. Micro-computed tomography measurements were validated against measurements from histological sections. The average thickness of the myometrium was found to be 0.33 ± 0.11 mm and 0.31 ± 0.09 mm in the left and right horns, respectively. The micro-computed tomography and histology thickness calculations were found to correlate strongly at different locations in the uterus: at the cervix, r = 0.87, and along the horn from the cervical end to the ovarian end, respectively, r = 0.77, r = 0.89 and r = 0.54, with p < 0.001 in every location. This study shows that micro-computed tomography can be used to quantify the musculature in the whole non-pregnant uterus and can be used for 3D virtual histology.

Prolonged stretching of rat uteri causes hypoxia and inhibits contractility via potassium channel TREK1.

During pregnancy, uterine kept quiescence along with uterine overdistention before labor. Prolonged stretching induced uterus myometrial hypoxia, increased TREK1 expression, and relaxed the myometrium, which may contribute to uterine quiescence and atony during pregnancy. The mechanisms underlying pre-labor uterine quiescence and uterine atony during overdistention are unclear. TREK1 (a two-pore domain potassium channel) and hypoxia-inducible factor-1α (HIF-1α) are activated by mechanical stretch, and their expression is upregulated by decreased uterine contractility. HIF-1α is a nuclear factor which regulates numerous target proteins, but whether it regulates TREK1 during the uterine stretch to cause uterine quiescence and/or atony is unclear. We investigated uterine contractility at different gestational stages in rats, as well as in non-pregnant uteri, which were induced by prolonged stretching and hypoxia. We also assessed the effects of incubating the uteri with or without echinomycin or l-methionine. Moreover, we analyzed HIF-1α and TREK1 expression levels in each group, as well as at various gestational stages of pregnant human uteri. We found that contractility was significantly decreased in pregnant uteri when compared with non-pregnant uteri, and this decrease was associated with increases in HIF-1α and TREK1 expression levels. HIF-1α and TREK1 expression levels in human uteri increased with the gestational length. Decreased uterine contractility and increased HIF-1α and TREK1 expression levels were also observed in non-pregnant rat uteri under 8 g of stretching tension or hypoxia. Inhibition of hypoxia with echinomycin restored normal uterine contractility, while HIF-1α and TREK1 protein expression remained reduced. TREK1 inhibition with l-methionine also restored uterine contractility under tension or hypoxia. In conclusion, we demonstrated that prolonged stretching induces myometrial hypoxia, increases TREK1 expression, and relaxes the myometrium, which may contribute to uterine quiescence and atony.

The Contractile Response to Oxytocin in Non-pregnant Rat Uteri Is Modified After the First Pregnancy

During pregnancy, the uterus undergoes several modifications under the influence of hormonal and mechanical stimuli. We hypothesize that while most of these modifications are reverted during involution, some of the physiological properties of the uterus are permanently altered. To investigate this hypothesis, we conducted motility experiments to evaluate the contractility response of uterine tissue samples from non-pregnant virgin and proven breeder female rats to oxytocin (10−10 to 10−5 M). We found that the virgin tissue contracts more robustly than proven breeder tissue in the absence of oxytocin, yet with oxytocin, proven breeder samples displayed a significantly higher increase in activity. These results could depend on a more elevated expression of oxytocin receptor and/or on an alteration in the intracellular pathways affected by the activation of the oxytocin receptors. Here, we explored the impact of some structures involved in the management of intracellular calcium on the dose response to oxytocin recorded from virgin and proven breeder uterine strips. Specifically, we replicated the dose response experiments in low extracellular calcium (10 μM), in the presence of the intracellular calcium channel blocker ruthenium red (10 μM), and in the presence of the sarcoplasmic-endoplasmic reticulum calcium ATP-ase pump inhibitor, cyclopiazonic acid (10 μM). The results of these experiments suggest that also the expression of proteins that control intracellular calcium availability is affected by the experience of pregnancy. Molecular biology experiments will give us more detail on the magnitude of these expression changes.

PS-B12-10: CLASSIC CARDIOVASCULAR PROTECTIVE SIGNALING IN ANGIOTENSIN II-INDUCED NOVEL PREECLAMPSIA RAT MODEL

Angiotensin II (Ang II)-induces vasoconstriction, in vivo perfusion has been shown to induce preeclampsia (PE) in pregnant rats. In this study, we aimed to examine the protein and mRNA expressions of endothelial and neuronal NOS (eNOS, nNOS) as well as angiotensin II type 2 receptor (AT2R) in normal pregnant and Ang II-induced PE rats. Pregnant (8 weeks) and non-pregnant SD rats (8 weeks, GD-8) were infused with Ang II via an osmotic minipump (1 ug/kg/min), and the rats (GD-21) were sacrificed 13 days later. Pregnant and non-pregnant rats without Ang II were taken as sham controls. The fetal weight, crown-rump length and placental weight of PE rats were significantly lower compared to those without Ang II infusion. Immunohistochemistry results showed an approximately 30% increase in glomerular volume, suggesting morphological changes in the kidney and Ang II infusion exhibits a PE phenotype. Ang II infusion did not affect AT2R, AT1R and nNOS protein or mRNA expression in the uterus of non-pregnant or pregnant rats. However, eNOS mRNA expression was reduced in PE rats (p &lt; 0.0001) although protein expression was not affected by Ang II infusion. However, in the placenta, AT1R and AT2R protein expression were not different in PE, but AT2R mRNA was decreased (p &lt; 0.0001). eNOS protein was only expressed in fetal vascular endothelial cells, and eNOS protein expression was higher in PE rats (P = 0.001) but eNOS mRNA was decreased (p = 0.0096). Notably, strong nNOS expression was observed in placental trophoblast cell groups of sham and PE rats, and both mRNA and protein expression of nNOS was decreased in placental lysates of Ang II rats (respectively p = 0.0287 and p &lt; 0.0001). These results demonstrate that we have recapitulated the PE model of Ang II infusion in rats. The expression of nNOS was reduced, and the localization of nNOS in the trophoblast suggested a new functional remodeling in the fetal vasculature. In the placenta, decreased expression of AT2R and nNOS may be a novel mechanism of impaired fetal development.

In vivo multi-channel measurement of electrical activity of the non-pregnant rat uterus.

In the uterus, the characteristics of smooth muscle contraction and the electrical activity that drives this contraction depends on hormonal cycles, and pregnancy status. Smooth muscle contraction is initiated by a change in membrane electrical potential, due to the flux of ions in and out of the intracellular space. Chains of action potentials throughout a section of muscle can result in coordinated contraction events. In this study, flexible printed circuit electrode arrays were applied to measure the bioelectric signals on the surface of a rat uterus in vivo. Variations in the electrical activity were quantified, including intermittent periods of activity and inactivity, which contain both slow-wave type activity (0.039 Hz ±0.017 Hz) and faster, spike-like activity (3.26 Hz ±0.27 Hz). The spike activity initiated at the ovarian end of the uterine horn, spreading towards the cervical end with a propagation velocity of 5.34 ± 2.32 mm [Formula: see text]. In conclusion, this pilot study outlines a new method of in vivo measurement of uterine electrical activity in rats. Clinical Relevance- Measurement of bioelectrical data using in vivo techniques provides insight into the electromechanical function of uterine smooth muscle, which could provide insights into what drives coordinated contraction in the uterus.

CLASSIC CARDIOVASCULAR PROTECTIVE SIGNALING IN ANGIOTENSIN II-INDUCED NOVEL PREECLAMPSIA RAT MODEL

Objective: Angiotensin II (Ang II)-induces vasoconstriction, in vivo perfusion has been shown to induce preeclampsia (PE) in pregnant rats. In this study, we aimed to examine the protein and mRNA expressions of endothelial and neuronal NOS (eNOS, nNOS) as well as angiotensin II type 2 receptor (AT2R) in normal pregnant and Ang II-induced PE rats. Design and method: Pregnant (8 weeks) and non-pregnant SD rats (8 weeks, GD-8) were infused with Ang II via an osmotic minipump (1 ug/kg/min), and the rats (GD-21) were sacrificed 13 days later. Pregnant and non-pregnant rats without Ang II were taken as sham controls. The fetal weight, crown-rump length and placental weight of PE rats were significantly lower compared to those without Ang II infusion. Immunohistochemistry results showed an approximately 30% increase in glomerular volume, suggesting morphological changes in the kidney and Ang II infusion exhibits a PE phenotype. Results: Ang II infusion did not affect AT2R, AT1R and nNOS protein or mRNA expression in the uterus of non-pregnant or pregnant rats. However, eNOS mRNA expression was reduced in PE rats (p &lt; 0.0001) although protein expression was not affected by Ang II infusion. However, in the placenta, AT1R and AT2R protein expression were not different in PE, but AT2R mRNA was decreased (p &lt; 0.0001). eNOS protein was only expressed in fetal vascular endothelial cells, and eNOS protein expression was higher in PE rats (P = 0.001) but eNOS mRNA was decreased (p = 0.0096). Notably, strong nNOS expression was observed in placental trophoblast cell groups of sham and PE rats, and both mRNA and protein expression of nNOS was decreased in placental lysates of Ang II rats (respectively p = 0.0287 and p &lt; 0.0001). Conclusions: These results demonstrate that we have recapitulated the PE model of Ang II infusion in rats. The expression of nNOS was reduced, and the localization of nNOS in the trophoblast suggested a new functional remodeling in the fetal vasculature. In the placenta, decreased expression of AT2R and nNOS may be a novel mechanism of impaired fetal development.

The ontogeny of kisspeptin receptor in the uterine contractions in rats: Its possible role in the quiescence of non-pregnant and pregnant uteri

The objectives of this study were to investigate the effects of KISS1 94-121 fragment on the contractility of non-pregnant and pregnant rat uteri, and to determine the uterine and myometrial expressions of Kiss1r. Uterine muscle strips were obtained from non-pregnant Sprague-Dawley rats in oestrous phase and from pregnant rats on gestational days 5, 15, 18, 20 or 22. The in vitro contractility measurements were carried out in an isolated organ bath in the presence of KISS1 94-121. Experiments with 5-day pregnant tissues were also performed in the presence of kisspeptin-234 trifluoroacetate. The mRNA and protein expressions of Kiss1r were measured by RT-PCR and Western blot analysis, while localizations of receptors were defined by fluorescent immunohistochemistry. KISS1 94-121 induced a dose-dependent relaxation both in non-pregnant and pregnant intact and endometrium-denuded uteri. A gradual decrease was found in the uterine expressions of Kiss1r mRNA and protein towards the end of the gestational period, and it was further confirmed by the immunohistochemical results. The significant majority of Kiss1r is found in the myometrium, however the few endometrial Kiss1r also influences the uterine contractions. The relaxing effect of kisspeptin is continuously reduced towards the end of gestational period in parallel with the reduction of Kiss1r expression. Our results suggest a putative role of kisspeptin in the maintenance of uterine quiescence that may have significance in miscarriage or preterm contractions.

Пептидоглікан золотистого стафілокока змінює скоротливість міометрія невагітних щурів завдяки підвищенню внутрішньоклітинного вмісту кальцію

The work is devoted to the study of the effect of Staphylococcus aureus peptidoglycan on the main parameters of myometrial contractions and the mechanism of this influence. The research was carried out by tensometry and calcimetry methods. The results of the experiments demonstrated that peptidoglycan modulates all the main parameters of contractions of the non-pregnant rat uterus, including increasing their amplitude and duration by 10 and 30%, respectively, mainly by prolonging the relaxation phase (56.7 ± 1.6%): myometrial band relative to quickly shrunk and relaxed for a long time. Application of peptidoglycan on freshly isolated uterine myocytes for 5 min led to a periodic increase in their intracellular content of calcium ions. Peptidoglycan enhanced myometrial contractions caused by hyperpotassium solution at a concentration of 60 mmol. Administration of 1 μmol of nifedipine, a blocker of potential-directed L-type calcium channels, suppressed spontaneous contractions enhanced by peptidoglycan. These changes in myometrium contractility of on the background of peptidoglycan action occur not only due to increased transmembrane calcium intake, but also its mobilization from the sarcoplasmic reticulum.

Relaxant effect of diallyl sulfide on nonpregnant rat uterus: Involvement of voltage-dependent calcium channels.

We aimed to determine the effect and mechanism of action of diallyl sulfide (DAS), an active component of sulfur-containing foods such as garlic on rat uterine activity. Isometric tension changes in longitudinal uterine strips obtained from 20 female Sprague-Dawley rats (250-300 g) in estrus stage of estrous cycle were studied in isolated organ baths containing Krebs-Henseleit solution. Diallyl sulfide (10-8 -10-6 M) caused a concentration-dependent relaxation on KCl (60 mM)-induced contractions and inhibited spontaneous peristaltic activity of uterine strips (P < 0.05). None of the following antagonists significantly changed the inhibitory effect of DAS on both KCl-precontracted uterine strips and spontaneous peristaltic activity of the uterus (P > 0.05): nitric oxide synthase inhibitor L-NAME (10-4 M), hydrogen sulfide-producing enzymes cystation β synthase and cystation γ-lyase inhibitors, aminooxyacetic acid (10-4 M) and propargylglycine (10-3 M) and nonselective cyclooxygenase inhibitor indomethacin (10-4 M). However, in calcium-free Krebs solution containing high KCl (30 mM), DAS significantly inhibited CaCl2 (10-5 -10-2 M)-induced uterine contractions in a concentration-dependent manner (P < 0.05). Diallyl sulfide has a relaxing effect on KCl-contracted rat uterus strips and an inhibitory effect on spontaneous uterine activity, possibly by decreasing the calcium influx into the cytoplasm of uterine smooth muscle cells.

Involvement of KАТР-channels of plasma and mitochondrial membranes in maintaining the contractive function of myometrium of non-pregnant rat uterus

Involvement of KАТР-channels of plasma and mitochondrial membranes in maintaining the contractive function of myometrium of non-pregnant rat uterus

Aqueous extract of Moringa oleifera leaf used in Nigerian ethnomedicine alters conception and some pregnancy outcomes in Wistar rat

The leaf of Moringa oleifera (Lam) Moringaceae, is globally accepted in ethnomedicine and particularly used to tackle malnutrition and enhance breast milk production in resource-limited settings. It is reported to be laden with a wide array of interesting bioactive plant metabolites. However, its safety at conception and during pregnancy has not been well reported notably in Nigeria. This work investigated the in vitro and in vivo uteroactive and contraceptive potentials of cold and hot aqueous extracts of M. oleifera leaves (MOL) commonly used in Nigerian Traditional Medicine. In addition, we determined the physicochemical characteristics of the freeze-dried aqueous extracts and powder.Powdered MOL (100 g each, 1:10 w/v) was extracted in the cold (room temperature) and hot (boiling) distilled water. The in vitro protocol involved the dose-dependent effect of aqueous extracts of MOL on the tension produced by the isolated uteri of non-pregnant rats using organ bath. Uterine contractility and contraceptive properties of MOL were assessed in vivo using female Wistar rats (150–170 g). Particle morphology, size, polydispersity index and zeta potential of MOL were determined.The in vitro administration of cold and hot aqueous extracts of MOL significantly (p < .05) produced uterine contractility with varying intensities. Contractile effect of cold MOL aqueous extract (89.7%) was higher than that of hot MOL (50.6%). In vivo, MOL cold aqueous extracts administered before and after mating, respectively produced 100% and 80% inhibition of conception, while 96.6% and 58% effect was comparatively produced by hot aqueous extracts of MOL.This work has provided useful data on MOL, a commonly consumed medicinal preparation/food supplement and the possible implications of its intake before conception, at conception and during pregnancy.

Pengaruh Pemberian Ekstrak Air Buah Pepaya Muda (Carica papaya.L) terhadap Gambaran Histologi Ovarium Tikus Betina (Rattus norvegicus)

Papaya seed (Carica papaya L.) as antifertility has been known since 1970 can cause the quantity and quality of sperm decreased. This antifertility potency is reinforced by the existence of research showing that inside papaya seeds contain antifertility compound that is saponin. In young papaya fruit (Carica papaya L.) also contains saponins, alkaloids, minerals, vitamins, and enzymes. The latex sap from the young papaya fruit has the same influence as oxytocin on the uterus of both pregnant and non-pregnant rats (Gouado et al., 2007) this study was conducted to prove that in the water extract of young papaya fruit has antifertility effect. If this study is meaningful, it may be used as a reference for subsequent research on efforts to utilize bio-natural resources as a medicinal substance.The purpose of this study is to know the influence of young papaya fruit extract on overview of ovarium histology of female rat.This research is a quantitative research using post-test design only control group design. This research is experimental with design. The research has been conducted from November to December 2017. This research was conducted at the Regional Center of Veterinary Investigation and Testing (RSVIT) II of Bukittinggi. The population in this study was female rats with a total number of female rats during the study of 32 rats.The results showed that Fresh Papaya Fruit Extract significantly influenced the number of primary follicles and follicles of rat Atresia (Rattus novergicus) (p = 0,000).Young Pepaya Fruit Water Extract can be considered as an efficient natural contraceptive and a few side effects.

Wine polyphenol resveratrol inhibits contractions of isolated rat uterus by activation of smooth muscle inwardly rectifying potassium channels

Resveratrol is a phytoalexin produced in a number of plant species including grapes. The benefit of resveratrol to health is widely reported. Resveratrol has been found to promote relaxation of non-pregnant and pregnant uterus, but its mechanism of action is unclear. The aims of our study were to investigate the involvement of inwardly rectifying potassium channels (Kir) in inhibitory effects of resveratrol on three models of contractions of non-pregnant rat uterus: the spontaneous rhythmic contractions (SRC), oxytocin-elicited phasic contractions and tonic oxytocin-elicited contractions.Uterine strips were obtained from virgin female Wistar rats in oestrus. Strips were mounted into organ bath for recording isometric tension in Krebs-Ringer solution. Experiments followed a multiple curve design. In order to test the involvement of Kirchannels in a mechanism of action of resveratrol(1-100 ?M),BaCl2 (1 mM),a antagonist of inwardly rectifying pota?ssium channels was used. Resveratrol induced a concentration-dependent relaxation of all models of contractions. BaCl2 antagonized the response to resveratrolon SRC and oxytocin-elicited phasic contractions. Relaxation achieved by resveratrolon tonic oxytocin-elicited concentrations was insensitive to BaCl2.The antagonism of resveratrol effects by inwardly rectifying potassium channels antagonist suggests that Kir channels are involved in resveratrol action on phasic contractions of rat uterus. Inhibitory effect of resveratrol on tonic contractions did not include Kir channels.

Oxytocin regulates the expression of aquaporin 5 in the late‐pregnant rat uterus

Aquaporins (AQPs) are integral membrane channels responsible for the transport of water across a cell membrane. Based on reports that AQPs are present and accumulate in the female reproductive tract late in pregnancy, our aim was to study the expression of AQP isoforms (AQP1, 2, 3, 5, 8, and 9) at the end of pregnancy in rat in order to determine if they play a role in parturition. Reverse-transcriptase PCR revealed that specific Aqp mRNAs were detectable in the myometrium of non-pregnant and late-pregnancy (Days 18, 20, 21, and 22 of pregnancy) rat uteri. The expression of Aqp5 mRNA and protein were most pronounced on Days 18-21, and were dramatically decreased on Day 22 of pregnancy. In contrast, a significant increase was found in the level of Aqp5 transcript in whole-blood samples on the last day of pregnancy. The effect of oxytocin on myometrial Aqp5 expression in an organ bath was also investigated. The level of Aqp5 mRNA significantly decreased 5 min after oxytocin (10(-8) M) administration, similarly to its profile on the day of delivery; this effect was sensitive to the oxytocin antagonist atosiban. The vasopressin analog desmopressin (3.7 × 10(-8) M), on the other hand, did not alter the expression of Aqp5, but did increased the amount of Aqp2 mRNA, an effect that was atosiban-resistant. These results lead us to propose that oxytocin selectively influences the expression of Aqp5 at the end of pregnancy, and may participate in events that lead to parturition in the rat. The sudden increase of AQP5 in the blood on the last day of pregnancy may serve as a marker that indicates the initiation of delivery.

In silico and in vitro pharmacological investigations of a natural alkaloid

Haplopine-3,3′-dimethylallyl ether (HAP) was isolated from a Sudanese herb (El-hazha), characterized and tested on isolated rat uterus rings pre-contracted with KCl by means of in vitro and in silico docking studies on a customized rat β2-adrenoceptor (β2-AR). The in vitro results revealed that HAP produces uterus-relaxant effects similar to those of terbutaline. Its E max (the maximum relaxing effect against KCl-induced contraction) was approximately 80% that of terbutaline, whilst its EC50 (The concentration producing 50% of E max) was ten times lower than that for terbutaline, showing greater potency and less efficacy. ICI-118,551 (a selective β2-AR antagonist) exhibited similar antagonism as concerns the relaxing effects of HAP and terbutaline, with pD 2 ′ = 6.3 ± 0.2 and pA 2 = 6.7 ± 0.2, respectively. The in silico results of β2-AR showed that HAP behaves similarly terbutaline except in the orientation within the active pocket. However, HAP has unique basic bond (His93) instead of the acidic bond at the most important site (Asp113) of β2-AR. This study suggests that the affinity is closely related to the interactions of the oxygen groups, whilst the difference in efficacy may be due to the lack of the Asp113 interaction. The correlation of the calculated ligand efficiency and E max revealed that there was a direct strong relation between the efficiency and in vitro efficacy. Finally, it was concluded that HAP can be categorized as a β2-AR agonist; further pharmacokinetic and toxicological studies are required to complete its profile. Left, Illustration of the lowest energy conformation of HAP (green) and the standard terbutaline (orange) docked into the active site of a customized rat β2-adrenoceptor, whilst right figure demonstrate dose–response curves of the relaxing activity of HAP (dashed line) and terbutaline (continuous line) on non-pregnant rat uterus in vitro against KCl-induced control contractions.

β 2-Adrenergic activity of 6-methoxykaempferol-3- O-glucoside on rat uterus: In vitro and in silico studies

6-Methoxykaempferol-3- O-glucoside (6-MKG) was isolated from a Sudanese herb (El-hazha). The pharmacological effects of 6-MKG were tested on isolated non-pregnant or late-pregnant rat uteri in vitro, whilst docking studies were carried out modelling of the binding of 6-MKG to the rat β 2-adrenoceptor in silico. In vitro studies revealed that 6-MKG was able to relax both the non-pregnant and the late-pregnant uterine contractility with 50% of the E max of terbutaline, whilst the EC 50 for 6-MKG was at least half than that of terbutaline. The β 2-adrenoceptors antagonist 3-(isopropylamino)-1-[(7-methyl-4-indanyl)oxy]butan-2-ol(ICI118,551) competitively antagonised the relaxing effect of 6-MKG. Radioligand binding and cAMP studies confirmed the β 2-adrenoceptors agonistic property of the compound. In in silico docking studies, 6-MKG bound to rat β 2-adrenoceptors with low ∆G bind value (− 11.53 ± 0.06 kcal/mol) and it interacted with four residues of the active site (Asp 113, Asn 312, Cys 191and Tyr 316). It is concluded that 6-MKG exerts weak β 2-adrenoceptor agonistic activity and can be considered a natural compound with potential therapeutic significance in the field of premature pregnant uterine contractions and asthmatic problems.

Innervation of the rat uterus at estrus: A study in full‐thickness, immunoperoxidase‐stained whole‐mount preparations

The innervation of the nonpregnant rat uterus has been studied in histological sections, which contain only small samples of nerves and are unlikely to afford a complete picture of uterine innervation. Here we used whole-mount preparations of entire full-thickness uterine horns from nonpregnant rats in estrus to visualize autonomic or sensory nerves with peroxidase immunohistochemistry. Immunoreactivity was studied for tyrosine hydroxylase (TH)-labeled sympathetic nerves; vesicular acetylcholine transporter (VAChT), parasympathetic nerves; and substance P (SP) and calcitonin gene-related peptide (CGRP), sensory nerves. Neuropeptide Y (NPY) and nitric oxide synthase (NOS) identified more than one of these functionally distinct nerve types. Axons of all neurochemical classes entered the uterus at the mesometrium and innervated the uterine smooth muscle. The linea uteri, a dense band of longitudinal muscle opposite the mesometrium, contained more TH-, NPY-, CGRP-, and VAChT-immunoreactive axons than the remaining smooth muscle. Axons immunoreactive for NPY, SP, NOS, and VAChT formed a plexus near the circular muscle-endometrium interface. Rare TH- and NPY-immunoreactive axons and occasional CGRP-immunoreactive axons occurred close to uterine glands. Blood vessels had dense perivascular plexuses of TH- and NPY-containing axons and less dense NOS-, SP-, CGRP-, and VAChT-positive plexuses. The circular muscle plexus and glands were absent opposite the mesometrium. Uterine arterioles formed an interconnected network throughout the uterus. This article provides the first comprehensive description of the autonomic and sensory innervation of the nonpregnant rat uterus and will be a foundation for future studies on changes in uterine innervation caused by normal physiological or pathophysiological challenges.

Potassium channels opener pinacidil have multiple effects on KCl-elicited contractions of isolated non-pregnant rat uterus

The effects of K+ channel opener, pinacidil on contractions provoked by contraction-stimulating KCl were investigated on isolated uterus of non-pregnant rats in oestrus. Pinacidil produced a more potent inhibition of 20 mM KCl-elicited contractions (pD2 = 6.57 μM) than of 40 or 80mMKCl-elicited contractions (pD2=5.11 and 5.19 μM, respectively). Glibenclamide, a selective blocker of adenosine triphosphate (ATP)-sensitive K+ (KATP) channels, antagonized the pinacidil-induced inhibition of contractions elicited by 20 mM KCl in a competitive manner. However, the pinacidil-induced inhibition of contractions provoked by 40 and 80 mM KCl glibenclamide was unable to prevent them. Pinacidil ability to completely relax the non-pregnant uterus pre-contracted with K+-rich solution suggests that K+ channelindependent mechanism(s) also plays a part in its relaxant effect.

Decreased Expression of the Rat Myometrial Relaxin Receptor (RXFP1) in Late Pregnancy Is Partially Mediated by the Presence of the Conceptus1

The actions of relaxin are mediated by relaxin family peptide receptor 1 (Rxfp1). In pregnant mice, myometrial Rxfp1 expression decreases at term, coinciding with the highest level of circulating relaxin. This down-regulation in Rxfp1 in reproductive tissues has not been investigated in other pregnant animals, nor are the regulatory mechanisms known. In the present study, we examined Rxfp1 gene and protein expression in the nonpregnant, pregnant, and postpartum rat uterus. The potential effects of local conceptus-derived factors on Rxfp1 expression were then examined in unilaterally pregnant rats. Immunoreactive RXFP1 was predominantly detected in the circular smooth muscle layer in the myometrium and in the decidualized endometrium. Rxfp1 was expressed in the rat myometrium from early to midgestation at levels similar to those in nonpregnant rat myometrium, with a significant reduction in expression at both the transcriptional and translational levels during late gestation. In unilaterally pregnant rats, myometrial Rxfp1 was higher in the nongravid compared to the gravid uterine horn, demonstrating a local negative influence of the fetal-placental unit on Rxfp1 expression. In summary, the down-regulation in myometrial Rxfp1 expression at the end of gestation in the rat is partially mediated by the fetal-placental unit and is indicative of a functional withdrawal of relaxin. This may represent a novel mechanism for the activation of spontaneous uterine contractions at labor in this species.

Effect of the ethanolic extract of Nauclea latifolia (Family: Rubiaceae) on the isolated uterus of non-pregnant rats

The plant Nauclea latifolia has been reported to be used by traditional healers to arrest pre-term labour. The ethanolic extract of the root of N. latifolia was screened for activity via agonist–induced contractions of uterine smooth muscles in non-pregnant female albino rats. The extract, at 0.1 and 0.2 mg/ml (final bath concentration), was tested against oxytocin (4×10-5 to 8×10-2 I.U/ml: final bath concentration), acetylcholine (0.04 to 40 μg/ml: final bath concentration) and ergometrine (0.05 to 100 μg/ml: final bath concentration) induced contractions invitro. The effect of the extract was compared to that of (0.004 μg/ml: final bath concentration) salbutamol and (0.004 μg/ml: final bath concentration) atropine. Both concentrations of the extract significantly shifted the concentration response curves of oxytocin (P<0.01), acetylcholine (P<0.0001) and ergometrine (P<0.0001) to the right with a slight depression of the Emax. This shift was more with the 0.2 mg/ml concentration, thus suggesting the possibility of a dose dependent action. There was no statistical significant decrease in Emax by 0.1 mg/ml of the extract, while the 0.2 mg/ml produced a significant depression (P<0.05) of the Emax, which like salbutamol could not be overwhelmed by higher concentrations of oxytocin. Similarly a significant reduction of the Emax of acetylcholine induced contractions was produced by 0.2 mg/ml, while both concentrations (0.1 and 0.2 mg/ml) produced significant (P<0.0001) reduction in Emax of ergometrine. It can thus be concluded that N latifolia root extractreduces oxytocin, acetylcholine and ergometrine-induced uterine contractions. These inhibitions were non-competitive. The result indicates an anti-abortifacient property. Key words: Antagonism, Nauclea latifolia, root extract, uterine-contraction, uterus