Rabbit Ovary Research Articles

The Interstitial Gland as a Source of Pro- or Anti-Senescent Cells during Chinchilla Rabbit Ovarian Aging.

The aging ovary in mammals leads to the reduced production of sex hormones and a deterioration in follicle quality. The interstitial gland originates from the hypertrophy of the theca cells of atretic follicles and represents an accumulative structure of the ovary that may contribute to its aging. Here, reproductive and mature rabbit ovaries are used to determine whether the interstitial gland plays a crucial role in ovarian aging. We demonstrate that, in the mature ovary, interstitial gland cells accumulate lipid droplets and show ultrastructural characteristics of lipophagy. Furthermore, they undergo modifications and present a foamy appearance, do not express the pan-leukocyte CD-45 marker, and express CYP11A1. These cells are the first to present an increase in lipofuscin accumulation. In foamy cells, the expression of p21 remains low, PCNA expression is maintained at mature ages, and their nuclei do not show positivity for H2AX. The interstitial gland shows a significant increase in lipofuscin accumulation compared with the ovaries of younger rabbits, but lipofuscin accumulation remains constant at mature ages. Surprisingly, no accumulation of cells with DNA damage is evident, and an increase in proliferative cells is observed at the age of 36 months. We suggest that the interstitial gland initially uses lipophagy to maintain steroidogenic homeostasis and prevent cellular senescence.

Histomorphology of the ovaries of rabbits does during ovulation induced by the combined use of gonadotropins

Hormonal stimulation of folliculogenesis is a general method of increasing reproductive capacity in rabbits; however, studying the safety of the developed protocols remains an urgent scientific task. The use of hormonal agents is due to the physiological characteristics of rabbits, primarily coitus-dependent mechanisms of ovulation. Applying gonadotropin-releasing analogs depends on mature follicles ready for ovulation in the ovary. It, therefore, has limited effectiveness for rabbits of the “zero” cycle that have not yet given birth. Therefore, such females are recommended to use medications based on gonadotropins obtained from the serum of foal mares (eCG) or human chorionic gonadotropin (hCG). There is limited data on the combined use of gonadotropins, especially the long-term administration in different doses. Thus, the research aimed to conduct a morphological evaluation of rabbit's ovaries with combined serum and chorionic gonadotropins to stimulate folliculogenesis before artificial insemination to assess the safety of the proposed dosages and long-term administration. Stimulation of ovulation in rabbits of experimental groups was caused by the combined use of serum and chorionic gonadotropins (eCG – 400 IU; hCG – 200 IU) for experimental groups 1 and 2 at a dose of 40 IU and 24 IU, respectively. Females of the control group were induced to ovulate by subcutaneous injection of 0.2 ml of gonadotropin-releasing hormone analog. Ovaries were collected on the seventh day of gestation after the euthanasia of the rabbit with prior premedication. After dissection and macroscopic examination, the ovaries were fixed in a 10% aqueous solution of neutral formalin. Histological slides were made according to the general method by pouring in paraffin and subsequent staining with hematoxylin and eosin. The research has established that repeated (during five reproductive cycles) combined use of serum and chorionic gonadotropins has dose-dependent effectiveness and can cause dysfunction of ovaries and blood circulation. Pathological changes may acquire a systemic character with the manifestation of critical conditions for the health and life of rabbits. The stimulating effect of the combined use of eCG and hCG was detected at a dose of 40 IU, and the inhibition of folliculogenesis was observed at 24 IU. Since the premises for the hyperandrogenism were found microscopically, it was essential to investigate the level of androgens in the blood of rabbits after using a dose of 24 IU. At the same time, both doses of gonadotropins affect the reactivity of the endothelium, which is manifested by the presence of intravascular vesicles and changes in the rheological properties of blood.

Characterization and establishment of an immortalized rabbit ovary granulosa cell line for reproductive biology experiments.

Granulosa cells (GCs) are the key components of ovarian follicles and regulate the maturation, communication, growth, and development of oocytes. GCs have great potential as human therapeutic models and in livestock breeding. In this study, we established an immortalized cell line (Im-RGCs) by transforming primary rabbit granulosa cells (Pri-RGCs) with lentivirus-mediated simian virus 40 Large T (SV40LT). Morphologically, Im-RGCs were indistinguishable from Pri-RGCs and maintained intact cell structure as observed by H&E staining. Also, Im-RGCs exhibited no significant change in cell proliferation, viability, and growth. Furthermore, GC-specific markers, such as FSHR, StAR, CYP11A1, and CYP19A1, were examined by PCR, immunofluorescence, and Western blotting. ELISA and karyotype analysis showed that Im-RGCs can synthesize steroid hormones and maintain the normal number of chromosomes during the infinite passage. Furthermore, soft-agar cloning and nude mice tumorigenic experiments indicated the absence of any malignancy transformation in Im-RGCs. In conclusion, we successfully established the immortalized granulosa cell line of rabbit follicles that can be used for biological, animal husbandry, and female reproductive research.

Morphological assessment of ovaries during stimulation of ovulation in rab-bits with equine serum gonadotropin in different doses

Ovulatory stimulation of rabbits during artificial insemination requires the use of hormonal drugs to stimulate maturation of follicles, effective fertilization and full manifestation of female sexual function. For this purpose, gonadotropins or analogues of gonadotropin-releasing hormone are common in the practice of rabbit breeding. The latter have a selective effect on folliculogenesis and, under certain conditions, are not capable of inducing ovulation in rabbits of “zero” reproductive cycle, that is, those that have not given birth. Instead, gonadotropins, in particular serum gonadotropin obtained from foal mares (eCG), have a pronounced effect on folliculogenesis in animals. eCG is a glycoprotein hormone secreted by trophoblastic epithelial cells of fetal origin. In other species, except for horses, it is able to induce the release of both luteinizing and follicle-stimulating hormone. That is why eCG is widely used together with human chorionic gonadotropin to induce follicle growth and ovulation. However, there are data on complications and pathological conditions that may occur with long-term use of eCG, especially in genital tract. Therefore, the goal of our research was the morphological evaluation of the functional compartments of ovaries when using eCG to stimulate reproductive function of rabbits. Schemes of hormonal treatment of females in the conditions of a private rabbit farm, an experiment was conducted, which included the introduction of eCG in different doses (40 IU for animals of experimental group 1 and 25 IU for experimental group 2) during five reproductive cycles. Selection of material for histological studies, its fixation, production and analysis of tissue sections stained with hematoxylin and eosin were performed according to commonly used methods. The complex of morphofunctional changes in the ovaries of the rabbits of experimental groups corresponds to the pathomorphological picture of the syndrome of hyperstimulated ovaries. In our experiment, obtained changes were more pronounced at a dose of 40 IU, using which we additionally detected signs of increased immunoreactivity in body of rabbits, as evidenced by the presence of nodular lymphoid formations in outer theca of secondary and tertiary follicles. Thus, as a safe regimen for the use of eCG for ovulatory stimulation in rabbits, we recommend a dosage of 25 IU for 1–2 reproductive cycles.

Behavioral and Physiological Differences in Female Rabbits at Different Stages of the Estrous Cycle.

Estrus involves a series of complex physiological signs and changes in behavior before ovulation, which play a crucial role in animal reproduction. However, there have been few studies that evaluate behaviors during the different stages of estrus cycle in female rabbits. Therefore, more detailed information is needed on distinguishing the various stages of the estrous cycle. This study explored the behavioral and physiological differences at various estrous cycle stages in female New Zealand White rabbits. The continuous recording method was employed to record the daily behaviors of twenty postpartum female rabbits during the estrous cycle. Compared with the diestrus stage, the duration of foraging and drinking behavior in estrus decreased significantly, and the frequency of grooming and biting behaviors increased (p < 0.05). Differences in reproductive hormone levels (FSH, LH, P4, and E2) and follicle development were measured at each stage via ELISA and HE staining. The FSH and LH levels showed an increasing trend and then decreased, with the lowest being in late estrus (p < 0.05). The P4 level was the lowest in estrus (p < 0.05), and E2 showed a gradually increasing trend. There was no significant difference in the number of primordial follicles at each stage, but the number of primary follicles in estrus was significantly higher than at the other stages (p < 0.05). To further understand the molecular regulation mechanism of the estrous cycle in female rabbits, we analyzed the ovarian transcription patterns of female rabbits in diestrus (D group) and estrus (E group) employing RNA-seq. A total of 967 differentially expressed genes (DEGs) were screened from the ovaries of female rabbits between the diestrus and estrus groups. A KEGG analysis of DEGs enriched in the estrogen signaling pathway, aldosterone synthesis, and secretion pathway, such as CYP19A1 and IGF1R, was performed. The rabbits' behavior, related physiological hormones, and molecular regulation also differed at different estrous cycle stages. The results provide recommendations for the adequate management practices of postpartum re-estrus and breeding female rabbits.

Histopathological effectiveness of vitex agnus castus in treating polycystic ovaries induced by letrozole in female rabbits

Female fertility is significantly impact by the common endocrine health issue known as polycystic ovarian syndrome (PCOS) in the childbearing years, Due to their ease of use and reduced cost-effectiveness when compared to chemical medicines generated via synthesis, therapeutic wild plants and herbs have been prized agents in the management of different illnesses for ages. Premenstrual syndrome, mastalgia, infertility, and menstrual cycle control are among the numerous conditions and diseases that can be treated with Vitex agnus-castus, a well-known medicinal plant that is native to the Mediterranean region and is widely used in Europe, Asia, and North Africa. The purpose of the study is to learn more about Vitex agnus catus and evaluate its efficacy in treating polycystic ovaries induced by letrozole in comparison to clomiphene citrate therapy. Since the effects of herbal activity are assessed by, observing histopathological changes in rabbit ovaries. The incidence of subcapsular ovarian cysts, capsular thickness, incomplete luteinization, and the quantity of corpora lutea were all higher in the research groups' ovaries. Compared to the group with a positive control (treated with normal saline only and administered orally). Histopathological study of the rabbit ovaries for both treatment groups revealed hyperplasia of the surface epithelium and congested blood vessels of the ovaries.

Light and focused ion beam microscopy workflow for resin-embedded tissues.

Although the automated image acquisition with the focused ion beam scanning electron microscope (FIB-SEM) provides volume reconstructions, volume analysis of large samples remains challenging. Here, we present a workflow that combines a modified sample protocol of the classical transmission electron microscope with FIB-SEM volume imaging. The proposed workflow enables efficient 3D structural surveys of rabbit ovaries collected at consecutive developmental stages. The precise trimming of the region of interest adds the time dimension to the volume, constructing a virtual 4D electron microscopy. We found filopodia-like processes emitted by oocyte cysts allowing contact between oocytes not previously observed.

Injection of Platelet Rich Plasma Intra-Ovarian to Treat Inactive Ovaries in Rabbits

Injection of Platelet Rich Plasma Intra-Ovarian to Treat Inactive Ovaries in Rabbits

Effect of light and dark shift on primary follicles diameter during post natal development in ovary of rabbit (Oryctolagus cuniculus)

Effect of light and dark shift on primary follicles diameter during post natal development in ovary of rabbit (Oryctolagus cuniculus)

Negative effects of heat stress on ovarian tissue in female rabbit.

Numerous studies have highlighted the role of miRNA in the deformation and necrosis of cells of ovarian tissue caused by heat stress (HS), which ultimately affects ovarian function. Although the role of small RNAs has been investigated in alterations in ovarian tissue functioning in response to HS, the expression profile of ovarian miRNA has been explored to a lesser extent. In this study, female rabbits were subject to HS treatment by using electrical heater. The current work demonstrated that HS could significantly change physiological performance of female rabbits including body weight, rectal temperature and relative ovary weight, and significantly reduce serum IL-2, IL-8, CAT, and GSH-Px concentrations by enzyme-linked immunosorbent assay (ELISA) technique. As a result, an increase in apoptosis in ovarian cells, as well as unhealthy follicles, were observed by Hematoxylin-eosin (HE) and TUNEL staining. Additionally, small RNA-seq revealed changes in the miRNA expression profile of rabbit ovaries under HS. Five hundred fourteen miRNAs were obtained including known miRNAs 442 and novel miRNAs 72. Among these miRNAs, 23 miRNAs were significantly expressed under HS. Eleven differentially expressed miRNAs (DE miRNAs) and 9 their predicted targets were confirmed by qPCR, which were expected miRNA-mRNA negative regulation pattern. Among the DE miRNAs and targets, miR-141-39 may target COQ6, miR-449a-5p and miR-34c-5p may control RFC5 and RTN2 together, miR-449a-5p may target ACADVL, miR-34c-5p potentially targets Bcl-2 and miR-196b-5p potentially regulates CASK and HOXB6. Thus, the current work suggested the negative effects of HS on the ovarian tissue of female rabbits, and in conclusion these changes could be caused by decreased serum IL-2, IL-8, CAT and GSH-Px levels, increased ovarian apoptosis, and changed the expression of miRNAs.

The effects of light colour on female rabbit reproductive performance and the expression of key genes in follicular development.

The purpose of this study was to analyse the effects of light colour on rabbit reproductive performance and the expression of key follicular development genes. Rabbits (n = 1,068, 5 months old, 3.6–4.4 kg live body weight) were divided randomly into four groups, housed individually in wire mesh cages and exposed to red, green, blue, and white light-emitting diode (LED) light (control). The lighting schedule was 16 L : 8 D-15 d / 150 lx / 6:00 am–22:00 pm (3 d preartificial insemination to 12 d postartificial insemination). Red light and white light affected the conception rate and kindling rate and increased the total litter size at birth (p < 0.05). The effects of red light on litter size at weaning, litter weight at weaning, and individual weight at weaning increased compared with the green and blue groups. The effects of red light on live litter size at birth were increased compared with those in the blue group (p < 0.05). Compared to white light, green and blue light reduced the number of secondary follicles (p < 0.05). Compared to red light, green and blue light reduced the number of tertiary follicles (p < 0.05). Compared with white light, red LED light resulted in greater ovarian follicle stimulating hormone receptor and luteinizing hormone receptor mRNA expression (p < 0.05). Compared with green and blue LED light, red LED light resulted in greater B-cell lymphom-2 mRNA expression (p < 0.05). Compared with green LED light, red LED light inhibited FOXO1 mRNA expression in rabbit ovaries (p < 0.05). Red light can affect the reproductive performance of female rabbits and the expression of key genes for follicular development.

Cryoprotective effect of antifreeze protein III on the rabbit ovary.

Ovarian tissue cryopreservation is effective in preserving fertility in cancer patients who have concerns about fertility loss due to cancer treatment. However, ischemia reduces the lifespan of grafts. Microvascular transplantation of cryopreserved whole ovary may allow immediate revascularisation, but the damage incurred during the cryopreservation procedure may cause follicular depletion; hence, preventing chilling injury would help maintain ovarian function. This study was designed to investigate the beneficial effects of antifreeze protein III (AFP III) on rabbit ovary cryopreservation. Ovaries (n =25) obtained from 5-month-old female rabbits (n =13) were frozen by slow freezing and vitrification. Cryoprotectant media were supplemented with and without 1mg/mL of AFP III. The experiment was divided into five groups: fresh control group (F), slow freezing group (S), slow freezing group with AFP III (AFP III-S), vitrification group (V) and vitrification group with AFP III (AFP III-V). All groups of ovaries were examined by histological characteristics analysis, ultrastructural analysis, apoptosis detection and follicle viability test. With slow freezing, the normal rate of change in follicle morphology, density of stromal cells and the survival rate of follicles in the AFP III supplemented group were significantly higher than those in the non-supplemented group, and a lower oocyte apoptotic rate was shown in the AFP III supplemented group. In the vitrification groups, the normal rate of change in follicle morphology and density of stromal cells in the AFP III supplemented group were significantly higher than those in the non-supplemented group, and a lower oocyte apoptotic rate was found in the AFP III supplemented group. But there was no obvious difference in the survival rate of follicles between the two groups. There was also no significant difference in the normal rate of change in follicle morphology, the survival rate of follicles and the apoptotic rate of oocytes between the vitrification and slow freezing groups (P >0.05), but the density of stromal cells in the vitrification groups was statistically higher than that of the slow freezing group (P <0.05). The addition of AFP III in slow freezing and vitrification could improve the cryoprotective effect of ovaries, which was more evident in slow freezing. The findings of this study provide a foundation for further research on the effects of AFP III in human ovarian tissue.

Effects of Dietary Polyphenols from Olive Mill Waste Waters on Inflammatory and Apoptotic Effectors in Rabbit Ovary.

Simple SummaryCircular economy strategies applied in the agro-food supply chain have recently been focused on new techniques for using agri-food by-products in animal nutrition. Rabbits efficiently convert plant proteins into foods rich in animal protein, therefore its meat, which contains numerous compounds, is potentially beneficial to human health and represents an excellent example of functional food. The residues obtained from the olive oil extraction process are a key source of phenolic compounds. In recent years, polyphenols have been used as dietary supplements to enhance animal health, welfare and performance, and to obtain functional foods of animal origin. Polyphenols enhance primary and total follicle number, and influence the inflammatory and apoptotic process by controlling and inhibiting pro-inflammatory cytokines (e.g., cyclooxygenase-2, interleukin-1beta, and tumor necrosis factor-alpha), and apoptotic factors (e.g., BCL2-associated X protein). Many health benefits of polyphenol consumption have not yet been determined, including the molecular and cellular mechanisms responsible for these actions at ovarian level. This paper highlights that dietary polyphenols, obtained from the olive oil industry, inhibit inflammatory and apoptotic activities in rabbit ovary, by modulating gene and protein expressions of cyclooxygenase-2 and BCL2-associated X protein, thus suggesting a direct involvement of these dietary compounds in mammalian reproduction.The aim of this study was to evaluate the effect of dietary polyphenols on the expression of the effectors involved in inflammation and apoptosis in rabbit ovary. New Zealand White female rabbits were fed a basal control diet (CTR), or the same diet supplemented with a polyphenolic concentrate (POL, 282.4 mg/kg) obtained from olive mill waste waters. The follicle counts and the relative mRNA (RT-qPCR) and protein (immunohistochemistry) expression of the effectors involved in inflammation (cyclooxygenase-2; interleukin-1beta; tumor necrosis factor-alpha, TNFA) and apoptosis (BCL2-associated X protein, BAX), detected in the ovaries of both groups, were examined. The POL diet increased the primary and total follicles number. Cyclooxygenase-2 gene expression was higher (p < 0.05) in the POL group than in the CTR group, whereas BAX was lower (p < 0.05) in POL than CTR. Immunohistochemistry revealed the presence of all the proteins examined, with weaker (p < 0.05) COX2 and BAX signals in POL. No differences between the CTR and POL groups were observed for IL1B and TNFA gene and protein expression. These preliminary findings show that dietary polyphenols modulate inflammatory and apoptotic activities in rabbit ovary, regulating cyclooxygenase-2 and BAX expression, thus suggesting a functional involvement of these dietary compounds in mammalian reproduction.

Effect of dietary supplementation with nettle or fenugreek on folliculogenesis and steroidogenesis in the rabbit ovary – An in vivo study

The objective of the study was to investigate the effect of dietary supplementation with nettle or fenugreek on folliculogenesis and steroidogenesis in the juvenile rabbit ovary. To gain insight into the mechanism of action of these herbs, we examined follicle formation, ovarian cell proliferation and apoptosis, steroidogenic enzyme abundance and steroid concentrations in ovarian tissue and plasma. Animals were fed with control, 1% nettle- or 1% fenugreek-supplemented pellets from 5 to 12 weeks of age (n = 10 per each group), when animals were slaughtered for ovary and blood collection. The addition of nettle decreased the numbers of primordial (P = 0.015) and early antral (P = 0.02) follicles and increased the number of primary (P = 0.04) ones when compared with the control group. Following fenugreek supplementation, the numbers of primary (P = 0.008) and antral (P = 0.027) follicles were greater, while the number of early antral (P = 0.003) follicles was lower in comparison with the control group. Nettle revealed apoptotic activity through activation of caspases 9 (P = 0.047), 8 (P = 0.022) and 3 (P = 0.004), whereas fenugreek increased (P = 0.042) follicular cell proliferation marked by PCNA protein abundance. Furthermore, only fenugreek targeted steroidogenic enzymes, decreasing CYP17A1 (P = 0.043) and increasing CYP19A1 (P = 0.048) protein abundances that resulted in enhanced estradiol biosynthesis and its elevated (P = 0.006) plasma concentration. In conclusion, both herbs affected follicle development in the rabbit ovary in a stage specific manner. Additionally, fenugreek altered ovarian steroidogenesis in a way that might affect sexual maturation in rabbits.

Гістологічне дослідження яєчників сукрольних кролиць

The objective of our experiment was to study the histrostructural changes in the ovaries of pregnant rabbits with previous reproductive cycles in artificial insemination without using further hormonal treatment to increase the reproductive function of rabbits. We have studied the ovaries of pregnant rabbits of the Hyla breed on the 7th day of pregnancy age, which were in the fifth reproductive cycle. The AI technique in rabbits needs an ovulation stimulator or inducer because female rabbits are induced ovulators. In our experiments, we used Surfagon for induction of ovulation by the method proposed by Lisin V. I., Sushko A.B., Institute of animal’s science of the NAAS. The ovaries were fixed in 10%-neutral formalin solution. Paraffin histosections were made and coloured with hematoxylin and eosin according to the standard technique. The macroscopic and microscopic studies of the rabbit ovary of the 5th reproductive cycles indicate a high level of their luteinization. The results of the histological study show that the histostructural picture of the ovaries of pregnant rabbits reflects the nature of the course of the phases of the previous reproductive cycles typical for this animal species. In the ovary of pregnant rabbits of the fifth reproductive cycle, all generations of follicles were found. The predominant functional elements of the ovarian parenchyma were luteal structures: the corpus luteum of pregnancy, atretic bodies and interstitial glandular tissue. There were two types of follicular atresia: obliterating, which took place in the preantral (primary and secondary) follicles and cystic and the antral tertiary follicles. The formation of hemorrhagic follicles was a type of cystic atresia. The formation of atretic bodies occurred as a result of luteinization of primary and secondary follicles. In the ovaries of pregnant rabbits of the fifth reproductive cycle, yellow bodies of various generations were found, that is the evidence of their functioning for several reproductive cycles. The number and development of corpus luteum in the right and left ovaries was asynchronous, which indicates the asynchronous nature of ovulation in the preceding and current reproductive cycles. Interstitial glandular tissue was the predominant histological element in the ovaries of rabbits of the fifth reproductive cycle. Compared with the ovaries of other mammalian species, it was found not only in the cortical, but also in the medulla of the ovaries of pregnant rabbits.

Efficiency of GnRH-Loaded Chitosan Nanoparticles for Inducing LH Secretion and Fertile Ovulations in Protocols for Artificial Insemination in Rabbit Does.

Simple SummaryNano-drug delivery systems can be employed for improving ovulation induction prior to artificial insemination (AI) in rabbits. In this study, different routes of administration and different doses of GnRH–loaded chitosan nanoparticles (GnRH–ChNPs) were assessed for inducing ovulation in rabbits, proving their usefulness to reduce the GnRH dose and animal handling and improving AI outcomes. The use of GnRH–ChNPs allows for the reduction of the conventional intramuscular GnRH dose to half without compromising fertility. However, the addition of GnRH–ChNPs to semen extenders, although successfully inducing ovulation, has negative impacts on fertility. Thus, more studies are needed to allow further adjustments.Gonadotropin-releasing hormone (GnRH)–loaded chitosan nanoparticles (GnRH–ChNPs) were used at different doses and routes of administration to induce ovulation in rabbits as an attempt to improve artificial insemination (AI) procedures and outcomes. In this study, the characteristics (size, polydispersity, loading efficiency, and zeta-potential) of GnRH–ChNPs and the GnRH release pattern were determined in an in vitro study. A first in vivo study assessed the pituitary and ovarian response to different GnRH–ChNPs doses and routes of administration (two i.m. doses, Group HM = 0.4 µg and Group QM = 0.2 µg, and two intravaginal doses, Group HV = 4 µg and Group QV = 2 µg) against a control group (C) receiving bare GnRH (0.8 µg). The HM, QM, and HV treatments induced an earlier LH-surge (90 min) than that observed in group C (120 min), whilst the QV treatment failed to induce such LH surge. The number of ovulation points was similar among treatments, except for the QV treatment (no ovulation points). A second in vivo study was consequently developed to determine the hormonal (progesterone, P4, and estradiol, E2) profile and pregnancy outcomes of both HM and HV treatments against group C. The treatment HM, but not the treatment HV, showed adequate P4 and E2 concentrations, conception and parturition rates, litter size, litter weight, and viability rate at birth. Overall, the use of GnRH–ChNPs allows for a reduction in the conventional intramuscular GnRH dose to half without compromising fertility. However, the addition of GnRH–ChNPs to semen extenders, although successfully inducing ovulation, has negative impacts on fertility. Thus, more studies are needed to explore this point and allow further adjustments.

Effect of Moringa oleifera (Lam) Leaf Extract on Mating Readiness and Development of Rabbit Ovarian Follicles

Moringa oleifera (Lam) is feed rich nutrients and bioactive compaunds including phytosterogen types namely phytosterols and isoflavones. The purpose of thid study was to test moringa leaf extracts with diferent solvents against mating readiness and ovarian foliccle development in rabbit does . Three moringa leaf extracts namely ethanol coarse extract (EKE) ethanol fraction extract (EFE) and hexan fraction extract (EFH) were used in this study. Seven treatments were tried on 28 rabbit does in the extracts in pellet feed at a dose of 0 % (without extract), EFH 0.13%, EFH 0.26%, EFE 0.37, EFE 0.74%, EKE 0.54% and EKE 1.08%. Each treatment is repeated on 4 tails rabbit does. The variables in this study were mating readiness and histology of ovarian follicle development. The results showed that the treatment of EFH administration of 0.26% caused 100% mating readiness and 75% pregnancy rate. The addition of EFH, EFE and EKE treatment led to a decrease in follicle development in the ovaries of lactation rabbit mothers on day 28. From this study it was concluded that the administration of moringa leaf extract increases blood estradiol, resulting in passionate livestock but has the effect of lowering follicle development in the ovaries of rabbits.

Effects of dietary supplementation with algae, sunflower oil or soybean oil on folliculogenesis in the rabbit ovary during sexual maturation

This study aimed to examine the effects of algae (rich in omega-3 fatty acids), sunflower oil (rich in omega-6 fatty acids) and soybean oil (rich in omega-6 fatty acids) on the entire folliculogenesis in juvenile and sexually mature rabbits. After weaning, rabbits were randomly divided into four experimental groups of 14 animals each. Control animals received non-supplemented pellets, while in the other groups, the pellets contained 1% marine algae, 3% sunflower oil or 3% soybean oil. Animals from each group were slaughtered at 12 weeks of age (n = 7 per group) or at 18 weeks of age (n = 7 per group). The ovaries were harvested and fixed for hematoxylin-eosin staining, immunohistochemical localization of PCNA and TUNEL assay. Algae-enriched diet markedly decreased the number of primordial and primary follicles, while addition of sunflower oil reduced the number of primary follicles in 12-week-old rabbits. The number of antral follicles was higher following algae supplementation, but lower after addition of soybean oil in that age group. Proliferating index was decreased following supplementation with algae and soybean oil in juvenile rabbits, whereas it was increased after addition of algae and decreased following vegetable oils in mature ones. Dietary PUFAs did not impact apoptosis in the rabbit ovary of both age groups. The obtained results suggest that PUFA-enriched diet regulate either early folliculogenesis or antral follicle development in rabbits that might influence reproductive performance as a consequence. It appears that observed effects are attributed to sexual maturity.

Subcutaneous grafting of ovaries from newborn rabbits into adult ovariectomized mice does not result in normal folliculogenesis

About 10% (6.1 million) of women in the United States ages 15–44 have difficulty becoming pregnant or staying pregnant. About 1/3 are fertility problems in women. Abnormal ovarian function is a major contributor. A greater understanding of ovarian function and especially follicular development would improve treatment. Follicles form when germ cells arrive in the primitive sex cords, which are derived from the coelomic epithelium. In most species, these cords fragment and the germ cells become covered by precursors of the granulosa cells. Thus primordial follicles form. Pools of these follicles are eventually recruited and undergo maturation; a series of functional and morphological transformations that lead to ovulatory follicles. In species like humans, cattle and pigs, follicles form during the embryonic phase of gestation, while maturation takes place after birth. In contrast, both phases occur postnatally in rabbits and makes them an excellent model to study folliculogenesis. Laboratory studies using non‐rodent species are logistically more difficult and even prohibitive. One alternative is ovarian grafting into immunocompromised mice. When adult ovarian tissue is transplanted from large animals into mice, some primordial follicles survive and undergo maturation. We investigated the rabbit model by transplantation of immature rabbit ovaries under the dorsal skin of adult ovariectomized NCr N ude mice (1 ovary / mouse). At 4 – 5 weeks, growth of the grafted ovaries was evident. Macroscopically, 50% of the grafts (5/10) had one or two follicle‐like structures on their surface. Histologically the structures were cysts lined by cuboidal cells and contained cells with the germ cell phenotype. In conclusion, ovaries grafted subcutaneously from newborn rabbits into ovariectomized adult mice developed cystic primordial follicles. Normal follicle formation and maturation did not occur.Support or Funding InformationMidwestern University intramural funds

Inferring lanosterol functions in the female rabbit reproductive tract based on the immunolocalization of lanosterol 14-demethylase and farnesoid beta-receptor

Female reproductive organs have de novo synthesis of cholesterol. Some sterol molecules, intermediaries in the cholesterol synthesis, have important paracrine/autocrine actions. Lanosterol binds to the farnesoid beta-receptor (FXRβ), a molecule widely expressed in the ovaries, suggesting that it may play a role in reproduction. Up to date, we know little about lanosterol functions across female reproductive organs. We described immunolocalized lanosterol 14-demethylase (LDM or CYP51A1), responsible for catalyzing the conversion of lanosterol in cholesterol, and FXRβ in the ovary, oviduct, uterus, and vagina of virgin and pregnant rabbits. In virgin rats, we found CYP51A1 and FXRβ immunoreactivity was found in all ovarian follicles, epithelial cells, stroma, and Graafian follicles. Also, the epithelium and stroma, as well as the smooth muscle of the oviduct, vagina, and uterus showed CYP51A1 and FXRβ immunoreactivity. In pregnant dams, we observed the presence of CYP51A1 and FXRβ immunoreactivity in the corpora lutea, giant uterine cells, and trophoblastic cells. The presence of CYP51A1 and FXRβ support that lanosterol participates in diverse reproductive processes, including follicular maturation, transport of gametes and zygote, implantation of blastocyst, lubrication, and contraction of the vagina, secretion of female prostate, and control of delivery mediated by pelvic muscles contraction.