Ovarian Structures Research Articles

IMPACT OF INTRAVAGINAL PROBIOTICS INOCULATION ON REPRODUCTIVE PERFORMANCE OF HOLSTEIN DAIRY CATTLE DURING TRANSITION PERIOD

In this study we tested the hypothesis that intravaginal infusions of lactic acid bacteria (LAB) around calving would improve uterine involution and reproductive performance of dairy cows. The probiotic culture used in this study was a mixture of 2 LAB strains isolated previously from the vaginal tract of healthy pregnant heifers. Forty pregnant 2nd lactation dairy cows 2 wk before the expected day of calving were assigned to one of 2 groups, first group was the control group (C) receiving 1 mL of media only, and the 2nd group was the treated group (T) receiving 1 mL of media enriched with probiotic bacteria at 1010 to 1012 cfu / treatment. Intravaginal infusions were performed once during wk −2, −1, +1, +2, +3, and +4 relative to parturition with probiotic bacteria. All cows were observed for reproductive performance and reproductive diseases until next conception using ultrasonography. The incidence of uterine infections in the multiparous cows were measured. Days open, conception rate at 1st, 2nd and 3rd insemination and repeat breeders (%) were calculated. Incidence of purulent and foul-smelling discharges on +1, +2, +3 and +4 wk relative to parturition and numbers of cows in the clean-up program were measured, cervix diameters (C.D) and uterine horn diameters (U.H.D) were measured at days (22-29) and days (38-45) postpartum. Ovarian structures and endometritis were determined. Results revealed that uterine and cervical involution of postpartum dairy cows were earlier in cows treated with LAB. Cows showed purulent and fowl smelling discharges are fewer in number in treated group than control, also data showed that percentage of cows in the cleanup program was higher in the control group (40%) than in the treated group (20%). The averaged insemination number per conception was lesser in the treated group (2.55) than control (4). The conception rate at 1st insemination and 3rd insemination was higher in the treated group (35%, 20%) than in control one (25%, 10%), and the %of repeated breeder cows was lower in the treated group (30%) than control (40%), finally days open in the treated group (116.4) was much lower than in the control (154.72). Therefore it can be conclude that, intravaginal treatment of transition dairy cows with a mixture of lactic acid bacteria (LAB) could improve uterine involution, lower the incidence rates of uterine infections, and improve the reproductive performance.

Icariin exerts a protective effect against d-galactose induced premature ovarian failure via promoting DNA damage repair

Icariin is one of the most common active ingredients in traditional Chinese medicine, while its function against Premature ovarian failure (POF) has not been explored. POF animal model was induced by d-galactose, and icariin at different doses was administered. Ovarian structure and follicle counting were observed via hematoxylin and eosin staining. The levels of serum hormones were measured by ELISA. Primary ovarian granulosa cells were cultured to compare the protective effects of icariin on cell aging, and DNA damage markers including γH2AX and 53BP1 were assessed by Western Blot. Administration of icariin promoted ovary/body weight, follicles numbers and fertility outcomes. In addition, icariin downregulated the levels of follicle stimulating hormone and luteinizing hormone, and upregulated the levels of estradiol and anti-Müllerian hormone. Icariin protected ovarian granulosa cells from d-galactose induced aging, with increased cell viability and lower endogenous β-galactosidase activity. The alterations of expression level of γH2AX and 53BP1 by icariin indicated that the protection is via promoting DNA damage repair. In this study we tested the biological function of icariin against the d-galactose induced POF. Our results demonstrated that icariin effectively attenuated ovarian injury via promoting DNA damage repair, suggesting that icariin can be developed as a protective agent against POF.

Potential of Croton menyharthii and Uvariodendron kirkii aqueous extracts as female Novel contraceptives.

Objective: The aim of the study was to validate the traditional fertility regulating claims by investigating the effect of root bark extract of both plants on ovarian and uterine structures.Materials and Methods: Twenty five mature normocyclic female winstar rats were used. Group 1 consisted of 5 animals that acted as control. Group 2 and 3 with 5 animals each; received 500 and 800mg/ Kg Croton menyharthii respectively on alternative days for 28 days through intra- abdominal gavage. Group 4 and 5 were treated in a similar manner but received Uvariodendron kirkii aqueous extract. Left ovaries and uterus were harvested and processed for histomorphology.Results: Croton menyharthii and Uvariodendron kirkii extracts caused a reduction in primordial and antral follicles, disrupted granulosa and theca cells with significant degeneration of ova. Â Croton menyharthii caused a disruption of uterine endothelial structure and loss of villi. Uvariodendron kirkii aqueous extract caused a significant uterine gland vacuolation and a thickened endothelial lining with an intact endothelial cell layer and its invaginations. Disrupted ovarian and uterine structure possibly led to compromised fertility and implantation.Conclusion: The study validates the traditional use of the plant in fertility regulation. We suggest further investigation on these potential plants to address the call for novel contraceptive drugs.Â

Effect of tamoxifen on the Notch signaling pathway in ovarian follicles of mice

ABSTRACTWe investigated the effect of tamoxifen (TAM) treatment on the Notch signaling pathway in mouse ovary. Mice were randomly divided into four groups. Control group A animals were untreated. Control group B animals were treated with the vehicle only. Animals of the 0.5 TAM group received 0.5 mg/day TAM. Animals of the 1.5 TAM group received 1.5 mg/day of TAM. TAM was injected subcutaneously for 5 days. Body weights were measured at the start and end of the experiment. Sections were stained using Crossman’s modified trichrome to examine general ovarian structure. Other sections were immunostained to demonstrate Jagged 1, Ki 67 and Notch 2. The TUNEL method was used to detect apoptosis. No significant differences in body weight or ovarian weight were found among the experimental groups. The number of primordial follicles was greater in the treatment groups than in the control groups, while the number of antral follicles and corpora lutea were reduced in the treatment groups. Cell proliferation rates were decreased by TAM treatment and cystic follicles were formed in the ovarian stroma. Notch 2 expression in the granulosa cells was increased following TAM administration, but no change was found in Jagged 1 expression. TAM administration suppressed follicular development and exhibited a negative effect on ovarian morphology. Our findings suggest that the Notch pathway participates in the action of TAM. We suggest that it may be useful to use Notch pathway regulators to adjust the effects of TAM on the ovary.

A Systematic Study on Reproductive Endocrine Function Recovery From Subcutaneous Ovarian Autotransplantation in Mice After 2 Weeks

BackgroundHeterotopic ovarian transplantation can not only restore the reproductive and endocrine function of animals but can also be studied with a specialized animal model. The aim of the study was to determine whether the reproductive endocrine function of the subcutaneously transplanted ovary was restored after 2 weeks. MethodsThe ovaries of 7-week-old mice were autologously transplanted into the back muscle. Fourteen days later, the ovarian structure was examined by hematoxylin and eosin staining. We continuously observed vaginal smears for changes in the estrous cycle. Estrogen and androgen concentrations were detected on the 14th day. The oocytes were collected and then used for in vitro maturation (IVM) and in vitro fertilization (IVF). ResultsThe cyclical estrous cycle changes were similar to those of the control group. There were no differences in the serum androgen and estrogen levels between the graft and control groups. The oocytes were able to develop into blastocysts after IVM and IVF. These results indicated that ovarian endocrine and reproductive function were restored within 2 weeks. ConclusionOur studies have shown that this ovarian heterotopic autotransplantation technique is able to restore steroidogenic and gametogenic functions at day 14 after transplantation. So far, the 14th day after transplantation is a landmark during the recovery from autologous heterotopic ovarian transplantation in the back of the mouse. This time point is the appropriate window to study heterotopic ovarian transplantation in mice.

Effect of using GnRH plus P4 and estradiol plus P4 on ovarian response of embryo production of local Pesisir Selatan cows in West Sumatera

The objective of this study to evaluate the effect of GnRH plus P4 and estradiol plus P4 on superovulation responses and to know the relation between ovarian activity of local Pesisir Selatan cows. The data were obtained from 4 local Pesisir cows superovulated on breeding centre in west Sumatera. The treatment in superovulation was using GbRH plus P4 and estradiol plus P4 and data were analyzed using SPSS16 to determine theeffect of treatment on ovarian structure and the the regression correlation. The result showed that using GnRH plus P4 was greater ovulate rate, transferable embryos and lower the percentage of degerate and unfertize (P<0.05) than using estradiol plus P4. Linear regression between dominant follicle, thae number of CL and the number of embryos in coefficient correlation Rfcl=0.950; Rfe=0.829 and Rcle=0.980, respectively. In conclution the using of GnRH plus P4 for FTAI is greater the response in embryos production of local Pesisir Selatan cows.

The maternal effects of heat shock on biological parameters and ovaries of Frankliniella occidentalis (Thysanoptera: Thripidae)

Maternal effects of heat shock are reported for some species of insects, but little is known about such effects in the western flower thrips (WFT) Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae). WFT is a pest of vegetables in greenhouses worldwide. It is susceptible to high temperatures in its natural environment and is controlled using heat treatment in China. WFT population growth is suppressed by a brief exposure to a high temperature of 40°C or 45°C in the laboratory. To explore the mechanism by which high temperatures suppress the growth of WFT populations, as well as the effects of multiple heat treatments on WFT, we recorded the duration of development and survival of immature WFT, and the sex ratio (female/male) and fecundity of F1, F2, F3 and F4 adult females that developed after a single heat shock, and those of F2 offspring after a double heat shock. We also recorded the longevity and ovarian structure of adult females of the treated generation (P) and their F1, F2 and F3 offspring after a single heat shock. In addition, we determined whether the effects of a heat shock on second instar nymphs and adults differed. The results indicate that exposure of the parental generation to 41°C or 45°C for 2 h significantly prolonged the duration of development, reduced survival of immature WFT and altered the sex ratio (female/male), longevity and fertility of their adult female offspring. The effects of a heat shock of 41°C persisted for two generations, whilst the effect of heat shock of 45°C persisted for three generations. In addition, double heat shocks had more pronounced effects than a single heat shock. Heat shock administered to second instar nymphs resulted in a decrease in the number of ovarioles, whilst a heat shock administered to adults resulted in ovariole deformity. The maternal effects of heat shock in terms of the biological parameters of WFT, structure and number of ovarioles, are critical in determining the suppression of the growth at high temperatures of WFT populations.

An objective volumetric method for assessment of ovarian follicular and luteal vascular flow using colour Doppler ultrasonography

Our goal was to develop an objective computer-assisted volumetric method of assessing vascular flow from colour Doppler ultrasound data of ovarian structures recorded by free-hand movement. We hypothesized that a vascularity index (ratio of the region of blood flood to the region of ovarian structure) obtained from the three-dimensional volumetric analysis would be more precise (less variable) than conventional two-dimensional analysis of single images in estimating the functional status of the preovulatory follicles and corpus luteum. Doppler ultrasound cineloops of water buffaloes (Bubalus bubalis; n = 22) ovaries were recorded daily from 12 h before GnRH treatment to four days after ovulation. Cineloops were processed using Fiji and Imaris software packages for segmenting the area (two-dimensional analysis) and the volume (three-dimensional analysis) occupied by the blood-flow and associated tissue to calculate the vascularity index. For volumetric measurement, all images in a cineloop were used (i.e., no a-priori selection of images) while for two-dimensional analysis, three images from the region with apparent maximum vascularity were selected. The volumetric method was verified with theoretical ellipsoidal volume of the follicle (r = 0.96 P < 0.01) or corpus luteum (r = 0.58 P = 0.02). The variability in the follicular vascularity index among animals was lower using the volumetric method than two-dimensional analysis (0.018 ± 0.002 vs 0.030 ± 0.005, P < 0.01), while the variability for CL vascularity was similar between methods (P = 0.23). An increase in the follicular vascularity index was detected at 12 h after GnRH treatment using both methods (two-dimensional: 0.030 ± 0.008, P < 0.01; three-dimensional: 0.016 ± 0.006, P < 0.02). Buffaloes that ovulated tended to have a greater increase in 3D vascularity index than non-responding buffaloes (P = 0.06); the two-dimensional method was not able to detect these changes. Using the three-dimensional method, a moderate positive correlation (r = 0.59; P = 0.02) was evident between the follicular vascularity index at 14–16 h after GnRH treatment and follicular diameter. In conclusion, an objective volumetric method for assessing relative ovarian blood flow changes was developed using Doppler ultrasound cineloops recorded by free-hand movement. The 3-dimensional method eliminates the need for a-priori selection of images and is more precise as a result of decreased technical variability.

Decellularization of the mouse ovary: comparison of different scaffold generation protocols for future ovarian bioengineering

BackgroundIn order to preserve fertility in young women with disseminated cancer, e.g. leukemia, an approach that has been suggested is to retransplant isolated small follicles within an ovarian matrix free from malignant cells and with no risk for contamination. The present study evaluates the first step to create a bioengineered ovarian construct that can act as growth-supporting tissue for isolated small follicles that are dependent on a stroma for normal follicular maturation. The present study used the intact mouse ovary to develop a mouse ovarian scaffold through various protocols of decellularization.Material and methodsPotential Immunogenic DNA and intracellular components were removed from whole mouse ovaries by agitation in a 0.5% sodium dodecyl sulfate solution (Protocol 1; P1), or in a 2% sodium deoxycholate solution (P2) or by a combination of the two (P3). The remaining decelluralized ovarian extracellular matrix structure was then assessed based on the DNA- and protein content, and was further evaluated histologically by haematoxylin and eosin-, Verhoeff’s van gieson- (for elastin), Masson’s trichrome- (for collagens) and Alcian blue (for glycosaminoglycans) staining. We also evaluated the decellularization efficiency using the mild detergent Triton-X100 (1%).ResultsSodium dodecyl sulfate efficiently removed DNA and intracellular components from the ovarian tissue but also significantly reduced the integrity of the remaining ovarian extracellular matrix. Sodium deoxycholate, a considerably milder detergent compared to sodium dodecyl sulfate, preserved the ovarian extracellular matrix better, evident by a more distinct staining for glycosaminoglycan, collagen and elastic fibres. Triton-X100 was found ineffective as a decellularization reagent for mouse ovaries in our settings.ConclusionsThe sodium dodecyl sulfate generated ovarian scaffolds contained minute amounts of DNA that may be an advantage to evade a detrimental immune response following engraftment. The sodium deoxycholate generated ovarian scaffolds had higher donor DNA content, yet, retained the extracellular composition better and may therefore have improved recellularization and other downstream bioengineering applications. These two novel types of mouse ovarian scaffolds serve as promising scaffold-candidates for future ovarian bioengineering experiments.

Human placenta-derived mesenchymal stem cells inhibit apoptosis of granulosa cells induced by IRE1α pathway in autoimmune POF mice.

Previous studies have shown that the ovarian failure in autoimmune-induced premature ovarian failure (POF) mice could be improved by the transplantation of human placenta-derived mesenchymal stem cells (hPMSCs); however, the protective mechanism of hPMSCs transplantation on ovarian dysfunction remains unclear. Ovarian dysfunction is closely related to the apoptosis of granulosa cells (GCs). To determine the effects of hPMSCs transplantation on GCs apoptosis, an autoimmune POF mice model was established with zona pellucida glycoprotein 3 (ZP3) peptide. It is reported that the inositol-requiring enzyme 1α (IRE1α) and its downstream molecules play a central role in the endoplasmic reticulum (ER) stress-induced apoptosis pathway. So the aim of this study is to investigate whether hPMSCs transplantation attenuated GCs apoptosis via inhibiting ER stress IRE1α signaling pathway. The ovarian dysfunction, follicular dysplasia, and GCs apoptosis were observed in the POF mice. And the IRE1α pathway was activated in ovaries of POF mice, as demonstrated by, increased X-box binding protein 1 (XBP1), up-regulated 78 kDa glucose-regulated protein (GRP78) and caspase-12. Following transplantation of hPMSCs, the ovarian structure and function were significantly improved in POF mice. In addition, the GCs apoptosis was obviously attenuated and IRE1α pathway was significantly inhibited. Transplantation of hPMSCs suppressed GCs apoptosis-induced by ER stress IRE1α signaling pathway in POF mice, which might contribute to the hPMSCs transplantation-mediating ovarian function recovery.

Altering duration of the presynchronization period in a long-term progestin-based estrus synchronization protocol for timed artificial insemination of beef heifers

An experiment was designed to evaluate the effect of extending duration of the presynchronization treatment in a long-term progestin-based estrus synchronization protocol. Heifers were assigned to either an 18 d (Day 0–18) or 14 d (Day 4 to Day 18) CIDR® treatment (1.38 g progesterone controlled internal drug release insert; Zoetis, Madison, NJ), with prostaglandin F2α (PG; 250 μg im cloprostenol sodium) administered 16 d after CIDR® removal (Day 34). Heifers at two locations (location one, n = 193; location two, n = 649) were assigned to treatment based on reproductive tract score (RTS; Scale 1–5) and body weight. Heifers that were assigned RTS 1 were not retained for the trial (n = 6). Estrus detection aids (Estrotect®) were applied at PG. Split-time artificial insemination (STAI) was utilized and AI performed based on expression of estrus at 66 h. Expression of estrus was defined as removal of ≥50% of the grey coating from the Estrotect® patch. Heifers that expressed estrus at 66 h were inseminated then and heifers that had not expressed estrus were inseminated at 90 h. Only heifers that failed to express estrus by 90 h received gonadotropin-releasing hormone (GnRH; 100 μg im gonadorelin acetate) at the time of AI. At location one, blood samples were collected at PG and AI (66 h or 90 h) from all heifers to determine E2 concentration by radioimmunoassay, and transrectal ovarian ultrasound was performed to detail ovarian structures on a subset of heifers (n = 73) at both time points. The proportion of heifers expressing estrus did not differ between treatments, either by 66 h (60%) or in total by 90 h (84%) after PG. Pregnancy rate to STAI did not differ between treatments (P = 0.3; 52%, 14-d CIDR®-PG; 50%, 18-d CIDR®-PG), or at the end of the 60 d breeding season (P = 0.2; 86%, 14-d CIDR®-PG; 82%, 18-d CIDR®-PG). No differences were detected in mean diameter of the dominant follicle at PG (P = 0.6; 10.9 ± 0.4 mm, 14-d CIDR®-PG; 11.0 ± 0.4 mm, 18-d CIDR®-PG) or at STAI (P = 0.3; 12.6 ± 0.4 mm, 14-d CIDR®-PG; 13.2 ± 0.4 mm, 18-d CIDR®-PG), nor were any differences observed between treatments in concentrations of E2 at PG (P = 0.8; 1.1 ± 0.19 pg/ml, 14-d CIDR®-PG; 1.1 ± 0.19 pg/ml, 18-d CIDR®-PG) or STAI (P = 0.6; 3.8 ± 0.19 pg/ml, 14-d CIDR®-PG; 3.6 ± 0.19 pg/ml, 18-d CIDR®-PG). These data indicate that duration of CIDR® treatment can be extended from 14 to 18 d, thus providing flexibility in scheduling without compromising reproductive outcomes.

Effects of a single trace mineral injection on body parameters, ovarian structures, pregnancy rate and components of the innate immune system of grazing Nellore cows synchronized to a fixed-time AI protocol

Two experiments evaluated the effects of injectable trace minerals (ITM) administered 30 d before artificial insemination (AI) on body weight (BW), body condition score (BCS), ovarian structures, pregnancy rate, and components of the innate immune system of grazing Nellore cows. In Exp. 1, 20 multiparous cows (BCS = 4.3 ± 0.4, scale 1 to 9; BW = 388 ± 35 kg) were kept on marandu-grass pasture (Urochloa sp.) and offered free-choice access to a trace mineral supplement. Cows were stratified by BCS and BW and randomly assigned to 1 of 2 treatments: subcutaneous injection (6 mL/cow) of saline solution (0.9% NaCl) or ITM (Multimin 90; containing 60, 10, 5, and 15 mg/mL of Zn, Mn, Se and Cu, respectively) administered 30 d (d -30) before AI (d 0). Pregnancy diagnosis was evaluated on d 30, BW and BCS on d -30, 0 and 30, ovarian structures on d 0 and 14, and blood samples collected on d -30, -26, -22, -17, -11, 0, 7, 14, 21 and 30 for analysis of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), haptoglobin, ceruloplasmin and progesterone (P4). In Exp. 2, 573 multiparous cows (BCS = 4.8 ± 0.5; BW = 400 ± 35 kg) received free-choice access to a trace mineral supplement and were assigned randomly to 1 of 2 treatments (ITM or saline injection 30 d before AI). In Exp. 1, ITM did not affect (P ≥ 0.12) BW, BCS, corpus luteum (CL) diameter and volume, pregnancy rate, and plasma concentrations of haptoglobin, ceruloplasmin and P4 compared to saline injection. Administration of ITM increased (P ≤ 0.008) plasma concentrations of SOD on d -22 and -17 and GSH-Px on d -26 compared to saline. In Exp. 2, ITM tended to increase the pregnancy rate compared to saline injection for cows with BCS < 5 (P = 0.09; 58.4 vs. 46.8 ± 6.5% respectively), but not for cows with BCS > 5 (P = 0.36; 71.8 vs. 67.6 ± 3.5% respectively). Therefore, a single ITM injection 30 d before AI did not alter the body parameters, ovarian structures and acute phase response of grazing Nellore cows, but increased plasma concentrations of antioxidant enzymes, and tended to improve pregnancy rates to AI in cows with BCS < 5.

Hard and soft X-ray imaging to resolve human ovarian cortical structures.

Laboratory and synchrotron X-ray tomography are powerful tools for non-invasive studies of biological samples at micrometric resolution. In particular, the development of phase contrast imaging is enabling the visualization of sample details with a small range of attenuation coefficients, thus allowing in-depth analyses of anatomical and histological structures. Reproductive medicine is starting to profit from these techniques, mainly applied to animal models. This study reports the first imaging of human ovarian tissue where the samples consisted of surgically obtained millimetre fragments, properly fixed, stained with osmium tetroxide and included in epoxydic resin. Samples were imaged by the use of propagation phase contrast synchrotron radiation micro-computed tomography (microCT), obtained at the SYRMEP beamline of Elettra light source (Trieste, Italy), and X-ray absorption microCT at the Theoretical Biology MicroCT Imaging Laboratory in Vienna, Austria. The reconstructed microCT images were compared with the soft X-ray absorption and phase contrast images acquired at the TwinMic beamline of Elettra in order to help with the identification of structures. The resulting images allow the regions of the cortex and medulla of the ovary to be distinguished, identifying early-stage follicles and visualizing the distribution of blood vessels. The study opens to further application of micro-resolved 3D imaging to improve the understanding of human ovary's structure and support diagnostics as well as advances in reproductive technologies.

Identifying Early Conception Using Ultrasound in Ovsynch Treated Postpartum Anestrus Gir Cows

The aim of this study was to detect early conception using transrectal ultrasonography in postpartum anestrus Gir cows treated with Ovsynch protocol (Group-I, n=11), keeping the same number of normal cyclic cows as control (Group-II, n=11). Ultrasonography examinations were performed using a real-time B-mode ultrasound scanner equipped with 7.5 MHz convex array transducer on day 11, 22 and 32 post-AI to monitor ovarian structures and uterine changes. Cent percent of anestrus animals responded to the Ovsynch protocol with clear signs of estrus. The conception rates at induced and second estrus in the treatment group were 45.45 and 33.33%, respectively, with an overall conception rate of 2 cycles post-treatment as 63.63%. In the control group, first service, second service and overall conception rates of 2 cycles were 36.36, 14.28 and 45.45%, respectively. The results obtained were better with the Ovsynch protocol. Ultrasound scanning on day 22 and 32 revealed visualization of the fluid and embryonic vesicle, respectively. Early embryonic death occurred in one cow out of six between days 22 and 32 post-AI in ovsynch treated group.

MON-228 FSH/AMH Ratio and Adipocyte Size Are Important Indicators for Ovarian Cyst Formation

High-fat, high-sugar (HFHS) diet induces a number of metabolic dysfunctions. Furthermore, there is evidence that diet-induced obesity correlates with diminished reproductive outcomes, though the mechanism remains unclear. We utilized a female, HFHS rat model to determine the effects of adiposity on ovarian morphology and hormonal balance. Because animals were on an ad libitum diet, we observed a gradient of obesity and adiposity, as well as ovarian dysfunction. We explored the distribution of these variables in order to establish a potential mechanism for reproductive dysfunction in our HFHS model. To determine the effect of a HFHS diet on ovarian structures, adiposity, and follicle-relevant hormones, female Sprague-Dawley rats were separated into two diet groups. The control group (n=14) received standard chow and water ad libitum while the HFHS group (n=16) received 60% of calories from fat, 30% sucrose solution, and water ad libitum. Weights were recorded every other day, and estrous cycles were monitored through daily vaginal cytology. During the course of the study, a fraction of animals ceased cycling. Terminal blood samples were taken on diestrus-2. AMH and FSH levels in the plasma were measured using ELISA. Ovaries and abdominal fat pads were collected and sectioned for histological analysis. At the conclusion of the study, HFHS rats gained significantly more weight than controls and displayed larger abdominal and gonadal fat pads. Rats were divided into three groups based on estrous cycle status: non-cycling (n=7), HFHS cycling (n=8), and control cycling (n=7). Non-cycling and HFHS rats displayed different distributions in adipocyte size relative to controls, indicating a shift towards larger cell size. On average, non-cyclers and HFHS had more cysts then controls, and non-cyclers had fewer corpora lutea compared to HFHS and controls. While AMH and FSH levels did not differ between groups, the ratio of FSH to AMH significantly correlated with follicular structures. The FSH/AMH ratio displayed strong negative correlations with total follicle number (r=-0.72, p<0.0005) and late antral follicles (r=-0.78, p<0.00005), also correlating positively with the number of cysts per section (r=0.74, p<0.001). Independently, adipocyte size was also predictive of cysts per section (r=0.49, p<0.02). Taken together, however, the FSH/AMH ratio and adipocyte size strongly predicted number of cysts per section in multiple regression. Our results suggest that morphological changes resulting from diet-induced obesity occur along a spectrum, modulated by hormonal balance and adiposity. These results establish the FSH/AMH ratio and adipocyte size as important markers for the onset of ovarian dysfunction, particularly cyst formation. In addition, the FSH/AMH ratio predicts follicle counts in the ovary, providing a combined measure for prediction of follicle status.

Spatial relationships of ovarian follicles and luteal structures in dairy cows subjected to ovulation synchronization: Progesterone and risks for luteolysis, ovulation, and pregnancy

Objectives were to determine relative ovary location of follicles, GnRH-induced corpora lutea (CL), and older CL present in ovaries as part of ovulation synchronization and their associations with progesterone concentration and risk for luteolysis, ovulation, and pregnancy. Cows were exposed to a timed artificial insemination (AI) program [GnRH-1-7 d-PGF2α (1 dose or 2 doses 24 h apart)-56 h after first or only dose of PGF2α-GnRH-2-16 h-timed AI at 72 ± 3 d in milk]. Blood was collected to assess progesterone when ovarian structures were mapped in 694 cows before GnRH-1 and before and 48 h after PGF2α and, in a subset of cows, size of CL (n = 599) and progesterone (n = 380) at 6 d after AI. Dominant follicles and CL in single-ovulating cows were detected more often in right than left ovaries (follicles before GnRH-1: 60.6% right and GnRH-2: 61.2% right; and CL before GnRH-1: 58.6% right and GnRH-2: 66.4% right). Dominant follicles in single-ovulating cows before GnRH-1 tended to be ipsilateral to the CL more often than contralateral (54.8 vs. 45.2%) with co-dominant follicles identified in both ovaries (19.3%). In response to GnRH-1 or GnRH-2, more left-ovary follicles ovulated contralateral to CL (left to right, 54.7%; right to left, 34.7%) than right-ovary follicles, but fewer left-ovary follicles ovulated ipsilateral to CL (left to left: 45.3%) than right-ovary follicles ovulated ipsilateral (right to right: 65.3%). Preovulatory follicles in single-ovulating cows before PGF2α tended to be detected more often ipsilateral than contralateral to CL induced by GnRH-1 (younger CL; 56.5 vs. 43.6%), but were of equal frequency ipsilateral or contralateral to older CL present before GnRH-1. Luteolytic risk was less in cows bearing co-dominant follicles in both ovaries compared with those in either right or left ovaries. Luteolytic risk in single-ovulating cows did not differ between ovaries. Luteolytic risk was greater for cows bearing older CL (86.5%) than for cows bearing younger GnRH-1-induced CL (65.3%) or both (79.6%). Pregnancy risk at 60 d after AI was or tended to be greater in cows having both CL types compared with either younger or older CL, respectively, partly because of greater embryonic loss in the latter 2 cases. More female calves tended to be carried in right horns when conception occurred after first service, whereas the opposite greater female frequency occurred in left horns after repeat services. Right-ovary dominance is evident before and after GnRH treatment.

Функция трансплантатов овариальной ткани после гипотермического хранения: значение состава среды инкубации

Ð’ N€Ð°Ð±Ð¾N‚е Ð?N€Ð¾Ð²ÐµÐ´ÐµÐ½ NN€Ð°Ð²Ð½Ð¸N‚елNŒÐ½N‹Ð¹ анализ моN€N„ологиN‡ÐµNÐoой NN‚N€NƒÐoN‚NƒN€N‹, NÐ½Ð´Ð¾ÐoN€Ð¸Ð½Ð½Ð¾Ð¹ N„NƒÐ½ÐoN†Ð¸Ð¸ и N„ол-лиÐoNƒÐ»Ð¾Ð³ÐµÐ½Ð·Ð° N‚N€Ð°Ð½NÐ?ланN‚аN‚ов Ð?оловозN€ÐµÐ»Ð¾Ð¹ и неонаN‚алNŒÐ½Ð¾Ð¹ оваN€Ð¸Ð°Ð»NŒÐ½Ð¾Ð¹ N‚Ðoани ÐoN€N‹N Ð?оNÐ»Ðµ гиÐ?оN‚еN€Ð¼Ð¸N‡ÐµNÐoого N…N€Ð°Ð½ÐµÐ½Ð¸N (ГХ) в NN€ÐµÐ´Ð°N… N€Ð°Ð·Ð»Ð¸N‡Ð½Ð¾Ð³Ð¾ NÐ¾NN‚ава. ДлN вN‹Ð?олнениN N€Ð°Ð±Ð¾N‚N‹ бN‹Ð»Ð¸ иNÐ?олNŒÐ·Ð¾Ð²Ð°Ð½N‹ NÐ»ÐµÐ´NƒNŽN‰Ð¸Ðµ меN‚одN‹: геN‚еN€Ð¾N‚оÐ?иN‡ÐµNÐoаN N‚N€Ð°Ð½NÐ?ланN‚аN†Ð¸N (иNNÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ðµ NN‚N€NƒÐoN‚NƒN€N‹ и N„NƒÐ½ÐoN†Ð¸Ð¸ оваN€Ð¸Ð°Ð»NŒÐ½Ð¾Ð¹ N‚Ðoани Ð?оNÐ»Ðµ ГХ); иммNƒÐ½Ð¾-N„еN€Ð¼ÐµÐ½N‚нN‹Ð¹ анализ (оÐ?N€ÐµÐ´ÐµÐ»ÐµÐ½Ð¸Ðµ ÐoонN†ÐµÐ½N‚N€Ð°N†Ð¸Ð¸ Ð?оловN‹N… гоN€Ð¼Ð¾Ð½Ð¾Ð² в Ð?лазме); моN€N„ологиN‡ÐµNÐoий (оN†ÐµÐ½Ðoа N†ÐµÐ»Ð¾NN‚-ноNN‚и NN‚N€NƒÐoN‚NƒN€N‹ N‚N€Ð°Ð½NÐ?ланN‚аN‚ов N‚Ðoани Ð?оNÐ»Ðµ ГХ). ЭÐoNÐ?еN€Ð¸Ð¼ÐµÐ½N‚алNŒÐ½Ð¾ доÐoазано, N‡N‚о оÐ?N€ÐµÐ´ÐµÐ»NNŽN‰Ð¸Ð¼ N„аÐoN‚оN€Ð¾Ð¼ N„оллиÐoNƒÐ»NN€Ð½Ð¾Ð³Ð¾ N€Ð°Ð·Ð²Ð¸N‚иN и NÐ½Ð´Ð¾ÐoN€Ð¸Ð½Ð½Ð¾Ð¹ N„NƒÐ½ÐoN†Ð¸Ð¸ N‚N€Ð°Ð½NÐ?ланN‚аN‚ов оваN€Ð¸Ð°Ð»NŒÐ½Ð¾Ð¹ N‚Ðoани N€Ð°Ð·Ð»Ð¸N‡Ð½N‹N… NN‚адий гиNN‚огенеза Ð?оNÐ»Ðµ ГХ NÐ²Ð»NÐµN‚NN NÐ¾NN‚ав NN€ÐµÐ´N‹ инÐoNƒÐ±Ð°N†Ð¸Ð¸. УNN‚ановлено, N‡N‚о Ð?оNÐ»Ðµ Ð?N€Ð¸Ð¼ÐµÐ½ÐµÐ½Ð¸N N„оNN„аN‚но-NÐ¾Ð»ÐµÐ²Ð¾Ð³Ð¾ бNƒN„еN€Ð° длN ГХ знаN‡Ð¸Ð¼Ð¾ NƒÐ¼ÐµÐ½NŒNˆÐ°Ð»NN N„оллиÐoNƒÐ»NN€Ð½N‹Ð¹ Ð?NƒÐ» и NÐ½Ð¸Ð¶Ð°Ð»Ð°NNŒ NN‚еN€Ð¾Ð¸Ð´Ð¾Ð³ÐµÐ½Ð½Ð°N N„NƒÐ½ÐoN†Ð¸N N‚N€Ð°Ð½NÐ?ланN‚аN‚ов ÐoаÐo Ð?оловозN€ÐµÐ»Ð¾Ð¹, N‚аÐo и неонаN‚алNŒÐ½Ð¾Ð¹ оваN€Ð¸Ð°Ð»NŒÐ½Ð¾Ð¹ N‚Ðoани. ПоÐoазано, N‡N‚о в N€ÐµÐ·NƒÐ»NŒN‚аN‚е гиÐ?оN‚еN€Ð¼Ð¸N‡ÐµNÐoого N…N€Ð°Ð½ÐµÐ½Ð¸N в NN€ÐµÐ´Ðµ, NÐ¾Ð´ÐµN€Ð¶Ð°N‰ÐµÐ¹ манниN‚ол, ÐoолиN‡ÐµNN‚во N„оллиÐoNƒÐ»Ð¾Ð² (N€Ð°Ð·Ð½Ð¾Ð¹ NN‚еÐ?ени зN€ÐµÐ»Ð¾NN‚и и желN‚N‹Ðµ N‚ела) и NÐ½Ð´Ð¾ÐoN€Ð¸Ð½Ð½Ð°N N„NƒÐ½ÐoN†Ð¸N Ð?оNÐ»Ðµ N‚N€Ð°Ð½NÐ?ланN‚аN†Ð¸Ð¸ оваN€Ð¸Ð°Ð»NŒÐ½Ð¾Ð¹ N‚Ðoани бN‹Ð»Ð¸ NN€Ð°Ð²Ð½Ð¸Ð¼N‹ N N‚аÐoовN‹Ð¼Ð¸ Ð?оNÐ»Ðµ N‚N€Ð°Ð½NÐ?ланN‚аN†Ð¸Ð¸ NÐ²ÐµÐ¶ÐµÐ²N‹Ð´ÐµÐ»ÐµÐ½Ð½Ð¾Ð¹ N‚Ðoани.Probl Cryobiol Cryomed 2019; 29(1): XXX–XXXÂ

Expression of the plasminogen system in the physiological mouse ovary and in the pathological polycystic ovary syndrome (PCOS) state

BackgroundThe fibrinolytic system and its inhibitors play a number of roles, apart from their function in blood haemostasis and thrombosis, namely in ovarian folliculogenesis and in ovulation. Plasminogen is converted to active plasmin at the time of follicular rupture through a decrease in plasminogen activator inhibitor-1 (PAI-1) and an increase in plasminogen activators. Oligo−/anovulation and follicle arrest are key characteristics of PCOS, but studies evaluating fibrinolytic/proteolytic markers within human or animal PCOS ovaries are lacking. We aimed to investigate and compare the expression and distribution of the plasminogen system markers in PCOS and control ovaries.MethodsA hyperandrogenised PCOS mouse model was used that mimics the ovarian, endocrine and metabolic features of the human condition. Immunohistochemistry and digital image analysis were used to investigate and compare fibrinolytic/proteolytic markers plasminogen, plasminogen/plasmin, tissue plasminogen activator, urokinase plasminogen activator and inhibitor PAI-1 in PCOS and control ovaries. Student’s t-test was used to compare data sets for normally distributed data and Wilcoxon-Mann Whitney test for non-normally distributed data.ResultsWe noted differences in the ovarian distribution of PAI-1 that was expressed throughout the PCOS ovary, unlike the peripheral distribution observed in control ovaries. Plasminogen was present in small follicles only in PCOS ovaries but not in small follicles of control ovaries. When we assessed and compared PAI-1 expression within follicles of different developmental stages we also noted significant differences for both the PCOS and control ovaries. While we noted differences in distribution and expression within specific ovarian structures, no differences were noted in the overall ovarian expression of markers assessed between acyclical PCOS mice and control mice at the diestrus stage of the estrous cycle.ConclusionsOur novel study, that comprehensively assessed the fibrinolytic/proteolytic system in the mouse ovary, showed the expression, differential localisation and a potential role for the plasminogen system in the physiological mouse ovary and in PCOS. Androgens may be involved in regulating expression of the ovarian plasminogen system. Further studies evaluating these markers at different time-points of ovulation may help to further clarify both physiological and potential pathological actions these markers play in ovulatory processes distorted in PCOS.

Gonad pathology and intersex severity in pelagic (Tilapia zilli) and benthic (Neochanna diversus and Clarias gariepinus) species from a pesticide-impacted agrarian catchment, south-south Nigeria

Pesticides are distributed to different degrees in surface water and sediment, thus, risks of toxicity and adverse impacts to physiology of resident species could be determined by their microhabitat (sub-habitat) associations, either water column or sediment. River Owan receives pesticide input from diffuse sources from adjacent farmlands. Surface water, sediment and fish samples [(Pelagic: Tilapia zilli (n = 92) and benthic: Neochanna diversus (n = 59), Clarias gariepinus (n = 68) were collected within the catchment area across seasons for 18-months (August 2016–January 2018) and measured for pesticide levels. Testicular and Ovarian tissue samples across the three species were also examined for pathological alterations. Individual pesticide concentrations in surface water and sediment exceeded international allowable limits, while concentration of pesticide residues in tissues of benthic species was higher compared to pelagic fish. Histopathological assessment revealed a higher incidence of ovarian disruption including atretic follicles, intersex and disorganization of ovarian structure in benthic (bottom-dwelling) fish (C. gariepinus and N. diversus) compared to pelagic (water-column) fish (T. zilli). Males benthic fish species also recorded more severe anomalies, compared to pelagic fish. The damages and anomalies observed in ovarian and testicular tissue indicate chronic responses to pollutant exposure, and implicates the elevation of pesticide concentrations in surface water and sediment above permissible limits. The higher incidence of anomalies recorded for benthic species compared to pelagic species, indicates greater risks of reproductive disruption and could be associated with the microhabitat preferences (water-column or sediment).

Can Mesenchymal Stem Cell Transplantation and Fennel Ingestion Alter the Histology of Senile Diabetic Rat Ovary?

Background: The relation between menopausal status and diabetes mellitus remains controversial. However, clinical trials suggest that menopause would speed the progression to diabetes. Reproductive tract dysfunction is a recognized consequence of diabetes obesity syndrome (DOS). Therefore, a possible association between obesity, diabetes mellitus and abnormalities of female reproductive function may occur. Nowadays traditional medicinal plants and spices are commonly used for their possible effects as hypoglycaemic, antidiabetic and folliculogenic agents instead of using synthetic drugs. Recent studies suggest that bone marrow stem cells (BMSCs) are promising grafts to treat a variety of diseases, including reproductive dysfunction.Aim of the work: The objective of this study was to evaluate the therapeutic implication of MSCs and fennel ingestion on ovarian diabetically induced senile rats.Materials & Methods: 24 senile female albino rats were used in this study. They were divided into 4 equal groups: control group, diabetic group, diabetic/MSCs group and diabetic/ MSCs /fennel oil group. Treatments were continued for 8 weeks, and ovarian samples were collected for histological, immunohistochemical and ultrastructure studies.Blood glucose levels were estimated and serum hormonal analyses were performed.Results: Induction of diabetes caused variable sizes of typical ovarian follicular cysts and elevation of blood glucose level. Stem cell transplantation showed some amelioration in the ovarian structure and blood glucose level. Administration of both MSCs and fennel oil had the ability to lower the blood glucose level and restore ovarian structure in diabetic rats.Conclusion: The use of MSCs in regenerative medicine and tissue engineering makes them a promising therapy to treat diabetes mellitus complications. Fennel essential oil could be given to diabetic patients to ameliorate diabetes in addition to its use as a fertility enhancing agent.