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The Journal of Reproduction and... Dec 2020Cotton rats (Sigmodon hispidus, CRs) are commonly used as animal models in biomedical research. However, the reproductive characteristics and ovarian development in the...
Cotton rats (Sigmodon hispidus, CRs) are commonly used as animal models in biomedical research. However, the reproductive characteristics and ovarian development in the CRs has not been widely investigated. We have previously shown that female CRs, in particular, show several unique phenotypes associated with the urogenital system, such as chronic kidney disease and pyometra. Our investigation revealed unique morphologies in CR ovaries, particularly in oocytes. Cotton rat ovaries at 6-8 weeks of age were obtained from the Hokkaido Institute of Public Health, and their sections analyzed by light microscopy and transmission electron microscopy. Although the general histology and folliculogenesis of CR ovaries were similar to those of other experimental rodents, multi-oocyte follicles (MOFs) and double nucleated oocytes (DNOs) were also observed. Although MOFs were found at all stages of follicular development, a greater frequency of MOFs was observed in the primary and secondary stages. However, DNOs tended to be frequently observed in primordial follicles. Almost all MOF oocytes and a few DNOs possessed a clear zona pellucida, expressed DEAD (Asp-Glu-Ala-Asp) box polypeptide 4 and Forkhead box protein 2, a representative marker of oocytes and follicular epithelial cells. Thus, our investigations revealed the unique phenotypes of the CR ovary. As MOFs and DNOs are occasionally observed in human patients with infertility, the CR would be a useful animal model to study for gaining a better understanding of folliculogenesis and oocytogenesis, as well as their abnormalities in humans and other animals.
Topics: Animals; Epithelial Cells; Female; Mice; Mice, Inbred C57BL; Mice, Inbred ICR; Oocytes; Ovarian Follicle; Ovary; Phenotype; Rats; Reproduction; Sigmodontinae; Zona Pellucida
PubMed: 32879182
DOI: 10.1262/jrd.2020-061 -
Reproductive Biology and Endocrinology... Aug 2010Ovarian surface epithelial cells are thought to be a precursor cell type for ovarian carcinoma. It has been proposed that an increased rate of ovarian surface epithelial...
BACKGROUND
Ovarian surface epithelial cells are thought to be a precursor cell type for ovarian carcinoma. It has been proposed that an increased rate of ovarian surface epithelial cell proliferation during ovulatory wound repair contributes to the accumulation of genetic changes and cell transformation. The proliferation of ovarian surface epithelial cells during ovulatory wound repair has been studied primarily using immunohistochemical staining of paraffin-embedded ovary sections. However, such analyses require complex reconstruction from serially-cut ovary sections for the visualization and quantification of the cells on the ovarian surface. In order to directly visualize the proliferation and organization of the ovarian surface epithelial cells, we developed a technique for immunohistochemical staining of whole mouse ovaries. Using this method, we analyzed cell proliferation and morphologic changes in mouse ovarian surface epithelial cells during follicle growth and ovulatory wound repair.
METHODS
Three-week old FVB/N female mice were superovulated by sequential administration of pregnant mare's serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG). Ten hours after hCG administration, mice were given 5-bromo-2-deoxyuridine (BrdU) and euthanized two hours after BrdU administration for ovary isolation. The levels of incorporated BrdU in the ovarian surface epithelial cells were measured by staining paraffin-embedded ovary sections and whole ovaries with the BrdU antibody. Re-epithelialization of the ovarian surface after ovulatory rupture was visualized by immunohistochemical staining with E-cadherin and Keratin 8 in paraffin-embedded ovary sections and whole ovaries.
RESULTS
We determined that active proliferation of ovarian epithelial surface cells primarily occurs during antral follicle formation and, to a lesser extent, in response to an ovulatory wound. We also demonstrated that ovarian surface epithelial cells exhibit a circular organization around the wound site
CONCLUSION
Whole ovary immunohistochemistry enables efficient and comprehensive three-dimensional visualization of ovarian surface epithelial cells without the need for laborious reconstruction from immunohistochemically-stained serial ovary sections.
Topics: Animals; Bromodeoxyuridine; Cell Proliferation; Chorionic Gonadotropin; Epithelial Cells; Female; Gonadotropins, Equine; Immunohistochemistry; Mice; Ovary; Ovulation; Superovulation; Wound Healing
PubMed: 20712898
DOI: 10.1186/1477-7827-8-98 -
Tissue Engineering. Part A Jul 2020Cancer survivorship has increased considerably, but common cancer treatments may threaten female reproductive health and fertility. In females, standard fertility...
Cancer survivorship has increased considerably, but common cancer treatments may threaten female reproductive health and fertility. In females, standard fertility preservation techniques include egg and embryo banking and ovarian tissue cryopreservation, but these methods are not suitable for all individuals. Emerging fertility preservation technologies include follicle growth and ovarian bioprosthetics. Although these platforms hold tremendous promise, they remain in the preclinical phase likely because of our inability to adequately phenocopy the complexity of the ovarian environment. The goal of this study was to use an established research archive of fixed human ovarian tissue established through the Oncofertility Consortium to better understand the dynamics and milieu of growing follicles within the human ovary. We performed a histological analysis of the immediate surroundings of primary and secondary stage follicles. We evaluated oocyte and follicle diameters of these growing follicles, analyzed their growth trajectories, and mapped their precise relationships to other stage follicles within a defined area. We also stratified our findings according to participant age and previous treatment history. Our results serve as benchmarks for follicles grown and provide insight into how follicles should be seeded spatially within bioprosthetic ovaries, potentially improving the efficacy and clinical translation of these emerging technologies. Impact statement Life-preserving cancer treatments have greatly increased survivorship. However, treatments often have off-target health consequences that threaten female reproductive health and fertility. Although several standard fertility preservation options exist, there is a constant need to explore and expand options for all populations follicle growth and ovarian bioprosthetics are new experimental procedures, which are currently limited to proof of concept. In this study, we analyzed human ovarian tissue from a deidentified biospecimen repository to characterize the growing follicle landscape with the ultimate goal of informing bioengineering practices. This spatial analysis pinpoints the geometry of growing follicles within the human ovary and provides a framework for paralleling this environment in platforms.
Topics: Cryopreservation; Female; Fertility Preservation; Humans; Ovarian Follicle; Ovary; Tissue Engineering
PubMed: 32598235
DOI: 10.1089/ten.tea.2020.0051 -
Stem Cell Reports Nov 2023Three-dimensional (3D) stem cell models of the ovary have the potential to benefit women's reproductive health research. One such model, the reconstituted ovary (rOvary)...
Three-dimensional (3D) stem cell models of the ovary have the potential to benefit women's reproductive health research. One such model, the reconstituted ovary (rOvary) self-assembles with pluripotent stem cell-derived germ cells creating a 3D ovarian mimic competent to support the differentiation of functional oocytes inside follicles. In this study, we evaluated the cellular composition of the rOvary revealing the capacity to generate multiple follicles surrounded by NR2F2+ stroma cells. However, the rOvary does not develop a surface epithelium, the source of second-wave pre-granulosa cells, or steroidogenic theca. Therefore, the rOvary models represent the self-assembly of activated follicles in a pre-pubertal ovary poised but not yet competent for hormone production.
Topics: Female; Humans; Ovary; Ovarian Follicle; Oocytes; Granulosa Cells; Epithelium
PubMed: 37890483
DOI: 10.1016/j.stemcr.2023.10.001 -
Nihon Hoshasen Gijutsu Gakkai Zasshi Nov 2022It has been reported that the placement of protective equipment in female pelvic radiography is predominantly inadequate compared with that of male. We analyzed the...
PURPOSE
It has been reported that the placement of protective equipment in female pelvic radiography is predominantly inadequate compared with that of male. We analyzed the actual situation of ovarian shielding by protective devices using X-ray, CT, and MR images obtained in the past, and evaluated the effectiveness of gonadal protection in female hip radiography.
METHODS
The ovaries were contoured in MR images and the pelvic bone was extracted by CT images. The MR/CT fusion images were created using a 3D workstation. The amount of physiological fluctuation in the ovarian location was measured. The fusion images in the ray-summation display were manually superimposed with the X-ray image, and the percentage of ovaries that could be shielded by the protective device was classified into four categories: (a) complete protection, (b) partial protection, (c) failure of protection, and (d) image retaking.
RESULTS
The mean and maximum ovarian fluctuations were 1.1 cm and 3.9 cm in the superior and inferior directions, respectively, and 0.7 cm and 2.0 cm in the left and right directions, respectively. The percentage of ovaries shielded was 18.9% for complete protection, 58.5% for partial protection, 15.1% for inadequate protection, and 7.5% for image retaking.
CONCLUSION
The effectiveness of gonadal protection is low because the protective device could not cover the entire ovary in about 80% of the female hip radiographs.
Topics: Male; Female; Humans; Radiation Protection; Radiography; Hip Joint; Gonads; Ovary
PubMed: 36184464
DOI: 10.6009/jjrt.2022-1314 -
Biology of Reproduction Jul 2014Granulosa cell formation and subsequent follicular assembly are important for ovarian development and function. Two members of the GATA family of transcription factors,...
Granulosa cell formation and subsequent follicular assembly are important for ovarian development and function. Two members of the GATA family of transcription factors, GATA4 and GATA6, are expressed in ovarian somatic cells early in development, and their importance in adult ovarian function has been recently highlighted. In this study, we demonstrated that the embryonic loss of Gata4 and Gata6 expression within the ovary results in a strong down-regulation of genes involved in the ovarian developmental pathway (Fst and Irx3) as well as diminished expression of the pregranulosa and granulosa cell markers SPRR2 and FOXL2, respectively. Postnatal ovaries deficient in both Gata genes show impaired somatic cell proliferation and arrested follicular development at the primordial stage, where oocytes are either enclosed by one layer of squamous granulosa cells or remain in germ cell nests/clusters. Furthermore, germ cell nests and primordial follicles are predominantly localized to the central region of the Sf1Cre; Gata4(flox/flox) Gata6(flox/flox) ovaries, where the boundary between the medulla and cortex is almost nonexistent. Lastly, most of the oocytes are lost early in development in conditional double mutant ovaries, which confirms the importance of normally differentiated granulosa cells as supporting cells for oocyte survival. Thus, both GATA4 and GATA6 proteins are fundamental regulators of granulosa cell differentiation and proliferation, and consequently of proper follicular assembly during normal ovarian development and function.
Topics: Animals; Down-Regulation; Female; GATA4 Transcription Factor; GATA6 Transcription Factor; Gene Deletion; Granulosa Cells; Mice; Oocytes; Ovarian Follicle; Ovary
PubMed: 24899573
DOI: 10.1095/biolreprod.113.117002 -
Journal of Visualized Experiments : JoVE May 2011Many organs depend on stem cells for their development during embryogenesis and for maintenance or repair during adult life. Understanding how stem cells form, and how...
Many organs depend on stem cells for their development during embryogenesis and for maintenance or repair during adult life. Understanding how stem cells form, and how they interact with their environment is therefore crucial for understanding development, homeostasis and disease. The ovary of the fruit fly Drosophila melanogaster has served as an influential model for the interaction of germ line stem cells (GSCs) with their somatic support cells (niche) (1, 2). The known location of the niche and the GSCs, coupled to the ability to genetically manipulate them, has allowed researchers to elucidate a variety of interactions between stem cells and their niches (3-12). Despite the wealth of information about mechanisms controlling GSC maintenance and differentiation, relatively little is known about how GSCs and their somatic niches form during development. About 18 somatic niches, whose cellular components include terminal filament and cap cells (Figure 1), form during the third larval instar (13-17). GSCs originate from primordial germ cells (PGCs). PGCs proliferate at early larval stages, but following the formation of the niche a subgroup of PGCs becomes GSCs (7, 16, 18, 19). Together, the somatic niche cells and the GSCs make a functional unit that produces eggs throughout the lifetime of the organism. Many questions regarding the formation of the GSC unit remain unanswered. Processes such as coordination between precursor cells for niches and stem cell precursors, or the generation of asymmetry within PGCs as they become GSCs, can best be studied in the larva. However, a methodical study of larval ovary development is physically challenging. First, larval ovaries are small. Even at late larval stages they are only 100μm across. In addition, the ovaries are transparent and are embedded in a white fat body. Here we describe a step-by-step protocol for isolating ovaries from late third instar (LL3) Drosophila larvae, followed by staining with fluorescent antibodies. We offer some technical solutions to problems such as locating the ovaries, staining and washing tissues that do not sink, and making sure that antibodies penetrate into the tissue. This protocol can be applied to earlier larval stages and to larval testes as well.
Topics: Animals; Dissection; Drosophila melanogaster; Female; Larva; Ovary; Staining and Labeling; Tissue Fixation
PubMed: 21610675
DOI: 10.3791/2537 -
Dong Wu Xue Yan Jiu = Zoological... Oct 2013This study compared the accumulation of Zn²⁺ and Cu²⁺ in the ovaries and ova of loaches under different concentrations of Zn²⁺ (1.00, 2.50 and 5.00 mg/L...
This study compared the accumulation of Zn²⁺ and Cu²⁺ in the ovaries and ova of loaches under different concentrations of Zn²⁺ (1.00, 2.50 and 5.00 mg/L respectively) and Cu²⁺ (0.10, 0.25 and 0.50 mg/L respectively). The results showed that both Zn²⁺ and Cu²⁺ accumulated in the ovaries, and that the relationship between accumulation and time was linear over 20 days of exposure. The accumulation of the metals in ovaries was closely related to the concentration of exposure in the solutions (P<0.05), and was obviously affected by the time and doses. However, the Cu²⁺ concentration was significantly higher than Zn²⁺ (P<0.05). The development level of ova in the ovaries also correlated with the concentration and exposure period in the Zn²⁺ and Cu²⁺ solutions. This study compared the accumulation of Znand Cu in the ovaries and ova of loaches under different concentrations of Zn (1.00, 2.50 and 5.00 mg/L respectively) and Cu (0.10, 0.25 and 0.50 mg/L respectively). The results showed that both Zn and Cu accumulated in the ovaries, and that the relationship between accumulation and time was linear over 20 days of exposure. The accumulation of the metals in ovaries was closely related to the concentration of exposure in the solutions (0.05), and was obviously affected by the time and doses. However, the Cu concentration was significantly higher than Zn(0.05). The development level of ova in the ovaries also correlated with the concentration and exposure period in the Zn and Cu solutions.
Topics: Animals; Copper; Cypriniformes; Female; Ovary; Ovum; Zinc
PubMed: 24115671
DOI: No ID Found -
Toxicological Sciences : An Official... May 2019Phthalates are synthetic chemicals with widespread human exposure due to their use as additives in consumer products. Phthalate diesters are hydrolyzed in the... (Comparative Study)
Comparative Study
Phthalates are synthetic chemicals with widespread human exposure due to their use as additives in consumer products. Phthalate diesters are hydrolyzed in the environment and in the body to monoesters that may be more toxic than the parent compounds. This study tested the hypothesis that adult mouse antral follicles, but not neonatal ovaries, are able to metabolize an environmentally relevant mixture of phthalates. Whole neonatal ovaries and isolated adult antral follicles from CD-1 mice were cultured in media treated with vehicle control or 0.1-10 µg/ml of a mixture composed of 35% diethyl phthalate (DEP), 21% di(2-ethylhexyl) phthalate (DEHP), 15% dibutyl phthalate (DBP), 15% diisononyl phthalate (DiNP), 8% diisobutyl phthalate (DiBP), and 5% benzylbutyl phthalate (BzBP). After 4 days of culture, media were subjected to high-performance liquid chromatography tandem mass spectrometry to measure the amounts of diester phthalates and monoester metabolites. Ovaries and follicles were collected to measure the gene and protein expression of the enzymes required for phthalate metabolism. Monoester metabolites for all phthalates except DiNP were detected in the media for both culture types at most doses. The long-chain phthalates (BzBP, DEHP, and DiNP) were metabolized less than the short-chain phthalates (DEP, DBP, and DiBP) compared with respective controls. Expression of metabolizing enzymes was observed for all treatment groups in both culture types. These data indicate that mouse ovaries are capable of metabolizing low doses of phthalates and suggest that metabolic capacity differs for follicles at different stages of development.
Topics: Activation, Metabolic; Age Factors; Animals; Animals, Newborn; Cells, Cultured; Environmental Pollutants; Female; Gene Expression Regulation, Developmental; Gene Expression Regulation, Enzymologic; Mice; Molecular Structure; Organ Culture Techniques; Ovarian Follicle; Ovary; Phthalic Acids; Risk Assessment
PubMed: 30768133
DOI: 10.1093/toxsci/kfz047 -
Journal of Ayub Medical College,... 2017Incidence of stress is on the rise in our daily life involving various neurobiological, endocrinological and behavioral changes. Hunger stress has a potent influence on...
BACKGROUND
Incidence of stress is on the rise in our daily life involving various neurobiological, endocrinological and behavioral changes. Hunger stress has a potent influence on mental, physical, and reproductive health by affecting the hypothalamic-pituitary gonadal axis.
METHODS
It was a laboratory based randomized control trial. Adult female mice (BALB-c strain) weighing 25-27 grams on first day of estrous cycle were taken in two groups (ten each). Group A was kept in normal environment of animal house for one month. Group B was given hunger stress by restricting the diet to about 50% per day for one month. Right ovaries of the animals were dissected out and observed for shape, color, and weight. Histological slides were prepared for the count of primary, secondary, and tertiary follicles.
RESULTS
Statistically significant decrease in animal and ovary weight with significant fall in ovarian follicles was observed.
CONCLUSIONS
Hunger stress affects the ovaries by reducing its weight and number of follicles.
Topics: Animals; Female; Hunger; Mice; Mice, Inbred BALB C; Ovary; Stress, Physiological
PubMed: 29330998
DOI: No ID Found