Ovarian Follicle Development Research Articles

Effect of Date Palm Pollen Supplementation on the Egg Production, Ovarian Follicles Development, Hematological Variables and Hormonal Profile of Laying Hens.

Simple SummaryDate palm pollen (DPP) is a natural product produced from male palm flowers and is used to improve ovulation and fertility in both women and men because it contains amino acids, fatty acids, flavonoids, saponins and sterols. Thus, this experiment investigated the effect of DPP supplementation on the oviposition rate, egg quality, ovarian follicle development, hematological variables and hormonal profile of laying hens. The findings indicated that the egg weight (EW), egg mass, albumen quality and laying rate of laying hens treated with DPP levels were significantly increased compared to those of the hens in the control group. The increased DPP levels significantly (p < 0.05) increased white blood cells (WBC), hemoglobin (Hb) and total antioxidant capacity (TAC), while H/L ratio significantly decreased. The increased DPP level significantly (p < 0.05) increased follicle-stimulating hormone (FSH) and luteinizing hormone (LH) concentration, ovary and oviduct weights compared to those of the control group.This experiment studied the effect of DPP supplementation in the laying hens’ diet on the ovarian follicle development, hematological variables and hormonal profile of laying hens. Eighty-four, 78-week-old, Lohman LSL hybrids layers were equally divided into four groups (4 groups × 7 replicates × 3 hens); hens in the 1st group were fed a basal diet (control), while those in the 2nd, 3rd and 4th groups, were fed on the same diet and supplemented with 1.25, 2.5 and 5.0 g DPP/kg diet. The results showed that the egg weight, egg mass and laying rate of laying hens treated with DPP levels were significantly increased compared to those of the hens in the control group. Egg weight, egg surface area, albumen quality and percentage of the yolk in treated hens significantly increased compared with controls. The increased DPP levels in laying hens‘ diet significantly (p < 0.05) increased WBC, Hb and TAC, while heterophil/lymphocyte (H/L ratio) significantly decreased. Additionally, the concentrations of FSH and LH and the weights of ovary and oviduct in the treated hens significantly (p < 0.05) increased compared with the control. In conclusion, the DPP supplementation in the hen diet significantly improved egg production, EW, H/L ratio, ovarian follicles, FSH and LH hormones concentrations.

Evaluating the Expression of Wnt Pathway Related Genes in Mouse Vitrified Preantral Follicles: An Experimental Study

Background:Wnt signaling pathway plays critical role in ovarian follicle development. Therefore, the aim of this study was to evaluate the effects of vitrification on the expression of Wnt pathway related genes in preantral follicles (PFs).Methods:Isolated PFs (n=982) of 14–16 day old female mice (n=45: 15 for each group) were divided into fresh (n=265), toxicity (n=272), and vitrified (n=265). The mRNA levels of Wnt2, Wnt4, Lrp5 and Fzd3 were evaluated by real-time PCR on the 2nd and 6th days of culture period. One-way ANOVA was conducted to analyze the data. Post hoc Tukey’s HSD was used for multiple comparisons and p-value less than 0.05 was considered statistically significant.Results:The developmental parameters of fresh PFs were significantly higher than those of vitrified (p<0.001). There were no differences between fresh and vitrified PFs on the 2nd day of culture (p<0.001). Wnt4 expression levels decreased significantly in vitrified groups compared with fresh ones (p<0.001). Fzd3 and Lrp expression levels increased significantly in vitrified groups compared with those in the fresh group on the 2nd day (p<0.001). On the 6th day of culture period, the expression levels of Wnt2 and Fzd3 increased significantly in vitrified group compared to those of fresh group (p<0.001). Moreover, the expression levels of Wnt4 and Lrp increased significantly in toxicity groups compared to those of the control group (p<0.001).Conclusion:Vitrification increase the expression levels of Wnt2, Lrp and Fzd3 genes of PFs during in vitro culture.

Stimulation of follicle growth and development during estrus in Ettawa Grade does fed a flushing supplement of different polyunsaturated fatty acids.

Background and Aim:Flushing with the manipulation of fatty acids, particularly polyunsaturated fatty acids, like linoleic and α-linolenic acids in the ration, is a strategy to raise the nutritional status of the female mammals to improve ovarian function and follicle development. This study was designed to investigate the effectiveness of flushing supplementation with different types of polyunsaturated and saturated fatty acids in stimulating follicle growth and development during estrus in Ettawa Grade does with a low initial body condition score (BCS ≤2).Materials and Methods:Eighteen Ettawa Grade does in the second parity, with an average body weight of 32.11±2.19 kg, were divided into three groups according to the fatty acid supplemented to their ration: (i) About 2.8% lauric acid flushing (group); (ii) 2.8% linoleic acid flushing (LA group); and (iii) 2.8% α-linolenic acid flushing (ALA group). The ration was formulated to be isocaloric (total digestible nutrient = 77%) and isonitrogenous (crude protein = 15%). The experiment was conducted for 35 days; that is, 14 days for acclimatization and synchronization of the estrous cycle and 21 days for fatty acid flushing until the appearance of the next estrus. A completely randomized design was applied.Results:According to the results, none of the different fatty acids in the ration affected the nutrient intakes, BCSs, average daily gains, and plasma glucose, cholesterol, and progesterone concentrations of the three groups of does. However, the BCSs (by 0.8-0.9) and the plasma cholesterol concentrations were higher after fatty acid flushing for 21 days than before the flushing period. The ALA group had the highest number of large-sized preovulatory follicles, whereas the LAURIC group had the highest plasma estradiol concentration during estrus. All three groups had similar plasma progesterone concentrations during estrus after fatty acid flushing.Conclusion:Flushing supplementation with 2.8% ALA from flaxseed oil gave the best results in terms of stimulating the highest number of large-sized preovulatory follicles in Ettawa Grade does.

Singleton and twin pregnancies in Red Maradi goats: changes in maternal serum biochemical composition and ovarian biometrics at mid gestation

Pregnancy involves some structural and physiological adjustments to achieve an optimal outcome for the fetus and its mother. The magnitude of these changes is influenced by number of fetuses in utero. This research was designed to compare serum biochemical and ovarian morphometric changes associated with singleton and twin pregnancies in Maradi goats. Using Richardson formula, 2.1 × [CRL (cm) + 17], twelve (12) ovarian and blood samples (7 single and 5 twin) from mid gestation (≈70-100 dGA) pregnant goats were purposively selected. Ovarian weights, ovarian diameters, serum biochemistry and levels of ovarian sex hormones were determined and analyzed using standard procedures. Twin pregnant goats had higher (p &lt; 0.05) left ovarian weights, average ovarian weight, left ovarian diameters, average ovarian diameter, serum calcium and inorganic phosphorus levels compared with singleton pregnant goats. There was no significant variation (p &gt; 0.05) in the mean crown-rump length, mean gestational age, mean fetal weight, right ovarian weights, right ovarian diameters, serum concentrations of sodium, potassium, chloride, urea, creatinine, AST, ALT, total protein, estrogen and progesterone between sinlgeton and twin pregnant Red Maradi goats. Ovarian sections from the twin bearing mid gestation goats had more growing follicles and fewer primordial follicles compared with the singleton bearing goats. Findings from this study indicate that twin pregnancy, which could be genetically programmed, has the tendency to affect ovarian follicles development, ovarian morphometrics and calcium metabolism in mid gestation in Red Maradi goats.

Bisphenol A and Its Analogues Exhibit Different Cytotoxic and Mitochondrial Dysfunction Potential in Human Granulosa Cells

Bisphenol A (BPA) is an environmental endocrine disruptor and has been strongly associated with the development of numerous diseases, including ovarian follicle development disorders. BPA is being replaced by structurally similar chemicals, such as bisphenol S (BPS), bisphenol F (BPF) and bisphenol AF (BPAF). However, the toxicity of these analogues in female reproduction is unclear. Here, we investigated the induction of cytotoxicity and mitochondrial dysfunction in the human granulosa cell line KGN by BPA and its selected analogues. We found that BPA and its analogues, especially BPAF, significantly reduced cell viability and caused cytotoxicity. Furthermore, we observed that BPA and BPAF significantly reduced mitochondrial function, including decreasing ATP generation, promoting ROS production and increasing intracellular Ca2+ levels. An oxidative-antioxidant imbalance was also detected after exposure to these chemicals. In contrast, the total antioxidant capacity was significantly reduced. To our knowledge, this is the first report on the evaluation of the potential of BPA and its analogues to induce cytotoxicity and mitochondrial dysfunction in ovarian granulosa cells. Our study revealed the possible mechanism of BPA and its analogues inducing granulosa cell damage and suggested that mitochondrial dysfunction may play an important regulatory role in bisphenol-induced follicular development disorders.

Aquaporin 4 expression is downregulated in large bovine ovarian follicles

Aquaporin channels (AQPs) are known to play an important role in the development of ovarian follicles through their function in water transport pathways. Compared to other AQPs, research on the role of AQP4 in female reproductive physiology, particularly in cattle, remains limited. In our previous study, gene chip microarray data showed a downregulation of AQP4 in bovine cystic follicles. This study was performed to validate the AQP4 expression level at the protein level in bovine follicles using immunohistochemistry, Western blotting, and immunoprecipitation assays. Immunostaining data showed that AQP4 was expressed in granulosa and theca cells of bovine ovarian follicles. The ovarian follicles were classified according to size as small ( 25 mm) in diameter. Consistent with earlier microarray data, semi-quantitative PCR data showed a decrease in AQP4 mRNA expression in large follicles. Western blot analysis showed a downregulation of the AQP4 protein in large follicles. In addition, AQP4 was immunoprecipitated and blotted with anti-AQP4 antibody in small and large follicles. Accordingly, AQP4 exhibited a low expression in large follicles. These results show that AQP4 is downregulated in bovine ovarian large follicles, suggesting that the downregulation of AQP4 expression may interfere with follicular water transport, leading to bovine follicular cysts.

The effect of agar substrate on growth and development of cryopreserved-thawed human ovarian cortical follicles in organ culture

ObjectiveTo preserve human ovarian tissue structure and improve follicular growth and survival during in-situ culture, various biomaterials are used. In this study we aimed to compare agar as a cultivation substrate with matrigel-coated insert in order to achieve an optimum system for in-situ human follicle culture. Study designFrozen-thawed human ovarian cortical tissues were cultured on either matrigel-coated inserts or agar-soaked substrates. The proportion of morphologically viable and degenerated follicles at different developmental stages, secreted hormonal levels, and apoptotic and proliferation gene expressions were compared between the cultured groups after 7-days of culture. ResultsThe follicular growth was not significantly different between the two cultured groups, although showing higher percentage of growing follicles in agar cultured group. The secreted hormonal levels didn’t have any difference between two cultured groups. Although the apoptotic gene expressions didn’t show any difference between the cultured groups, the apoptotic index was lower in agar cultured group. In addition, Ki67 gene expression, a proliferative marker, showed a significantly higher expression in agar cultured group. ConclusionBased on the results, agar is as suitable as matrigel-coated inserts for the survival and growth of follicles during culture. Therefore, agar can be an inexpensive alternative substrate for culturing frozen-thawed human ovarian cortical strips.

Effect of anti-müllerian hormone on the development and selection of ovarian follicle in hens

To elucidate the role of anti-müllerian hormone (AMH) in regulating the development of ovarian follicles in laying hens, the expressions of follicle-stimulating hormone receptor (FSHR), AMH receptor type 2 (AMHR2), steroidogenic-related genes steroidogenic acute regulatory protein (STAR), cytochrome P450 side-chain cleavage (CYP11A1), and 3β-hydroxysteroid dehydrogenase (HSD3B1) genes were measured from different sized follicles and granulosa cells. The results showed that the expressions of FSHR and AMHR2 genes were higher in small follicles and decreased after follicular selection. Oppositely, the expressions of STAR, CYP11A1, and HSD3B1 were significantly increased after follicular selection. It indicated that AMHR2 might mediate AMH suppression in the stimulating effects of follicle-stimulating hormone (FSH) on steroidogenic-related genes expression. To make sure the effects of AMH in this process, a total of 40 hens were treated (negative control, sham operation, 150 ng AMH/d or 300 ng AMH/d) for 25 d. We analyzed ovarian morphology, progesterone concentration in blood plasma, and the expressions of steroidogenic genes in ovaries and follicles. The AMH300 group had significantly lower weight of ovary and hierarchical follicles. Egg weight and ovary weight in AMH150 group were higher than those of sham operation and AMH300 groups, so did hierarchical follicles weight. The steroidogenic genes expressions showed an increase in ovarian tissue and the largest follicle of AMH150 and AMH300 groups. However, progesterone level in the blood was reduced by AMH injection with different concentrations. To further verify the above results, granulosa cells from 6 to 8 mm follicles were cultured with AMH (0, 5, 10, 20, 40, or 80 ng/mL). The results revealed that excessive AMH (80 ng/mL) exerted an inhibitory effect on progesterone synthesis and the expressions of STAR, CYP11A1, and HSD3B1. However, these genes expressions showed a significant increase in 20 ng/mL AMH-treated group. In summary, AMH inhibited the development of prehierarchical follicles in laying hens. The effects of AMH treatment with different concentrations on follicle development showed the follicle was selected by changing FSH responsiveness of prehierarchical follicles.

Ablation of TGFBR3 (betaglycan) in oocytes does not affect fertility in female mice.

Ovarian follicle development is regulated by locally produced TGFβ superfamily members. The TGFβ type III receptor (TGFBR3, or betaglycan), which regulates the actions of diverse TGFβ ligands, including inhibins, is expressed in different ovarian cell types. However, its functional roles in the ovary have not been investigated in vivo. Here, we ablated Tgfbr3 in murine oocytes using the Cre-loxP system. Oocyte-specific Tgfbr3 knockout (cKO) females were fertile, producing litters of similar size and frequency as controls. Their ovarian weights and histology were also normal. Though we confirmed efficient recombination of the floxed alleles, we did not detect Tgfbr3 mRNA in purified oocytes from superovulated cKO or control mice. These results challenge earlier observations of betaglycan protein expression in this cell type. Regardless, Tgfbr3 in the murine oocyte is clearly dispensable for female fertility.

The effect of different concentrations of cerium oxide during pregnancy on ovarian follicle development in neonatal mice

Cerium is a member of the rare metals group and widely used in drug delivery, gene therapy, molecular imaging and medicine. In this study, we investigated the effect of different doses of Cerium (IV) oxide (CeO2 ) during pregnancy on neonatal mice ovaries, as well as its effect on blood biochemical parameters. Thirty pregnant NMRI mice were divided into five groups: Control and 4 groups treated with CeO2 (10, 25, 80, 250 mg/kg.bw i.p) at the GD7 and GD14. The ovarian histological of neonatal (2 and 6 day-olds), as well as blood serum of neonates at 15-dpp were analyzed. Count of ovarian primordial follicles in neonates at 2 dpp showed a significant decrease in the groups treated with 80 and 250 mg/kg.bw doses of CeO2 . There was also a significant decrease in ovarian primordial and primary follicles in neonates at 6-dpp at 250 mg/kg.bw doses of CeO2 in the control (P < 0.05). There was no significant difference in serum levels of malondialdehyde and total antioxidant capacity between the experimental and control groups. Our results suggest that the effects of CeO2 on the ovarian tissue of neonatal mice during pregnancy may be dose-dependent.

Effect of eCG treatment on gene expression of selected matrix metalloproteinases (MMP-2, MMP-7, MMP-9, MMP-10, and MMP-13) and the tissue inhibitors of metalloproteinases (TIMP-2 and TIMP-3) in the chicken ovary

Several metalloproteinases (MMPs) are present and functional in the chicken ovary and regulate the extracellular matrix (ECM) during follicle development, ovulation, atresia, and regression. The regulation of the abundance of MMPs in avian ovarian follicles, however, is largely unknown. The aim of the present study was to examine effects of equine chorionic gonadotropin (eCG) on abundance of selected MMPs and relevant tissue inhibitors of MMPs (TIMPs) in the hen ovary. The MMP-2 and MMP-9 activity was also determined. Results indicated there were effects of eCG on abundances of MMP-2, MMP-7, MMP-9, MMP-10, MMP-13, TIMP-2, and TIMP-3 mRNA transcript and/or protein relative abundances in white, yellowish, small yellow, and the largest yellow preovulatory (F3-F1) ovarian follicles. The response to eCG depended on the stage of follicle development, layer of follicular wall, and the type of MMPs or TIMPs affected by eCG. Furthermore, there was a pause in egg laying when eCG was administered and there were morphological changes in the ovary following eCG treatment that were associated with alterations in MMP-2 and MMP-9 activity. In general, the results indicate that eCG, which has primarily follicle stimulating hormone (FSH)-like bioactivities, is a negative regulator of MMP abundance and activity in the largest yellow preovulatory follicles. Results from the present study indicate the gonadotropins, especially FSH, by the regulation of transcription, translation, and/or activity of proteins of the MMP system have effects on the mechanisms that underlie ECM remodeling and cell function throughout ovarian follicle development in the chicken ovary.

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.

Microfluidic Systems for Assisted Reproductive Technologies: Advantages and Potential Applications.

Microfluidic technologies have emerged as a powerful tool that can closely replicate the in-vivo physiological conditions of organ systems. Assisted reproductive technology (ART), while being able to achieve successful outcomes, still faces challenges related to technical error, efficiency, cost, and monitoring/assessment. In this review, we provide a brief overview of the uses of microfluidic devices in the culture, maintenance and study of ovarian follicle development for experimental and therapeutic applications. We discuss existing microfluidic platforms for oocyte and sperm selection and maintenance, facilitation of fertilization by in-vitro fertilization/intracytoplastimc sperm injection, and monitoring, selection and maintenance of resulting embryos. Furthermore, we discuss the possibility of future integration of these technologies onto a single platform and the limitations facing the development of these systems. In spite of these challenges, we envision that microfluidic systems will likely evolve and inevitably revolutionize both fundamental, reproductive physiology/toxicology research as well as clinically applicable ART.

Impact of acute short-term high thermal stress during early embryogenesis on hatchability, physiological body reaction, and ovarian follicles development of quails

This experiment aimed to evaluate the impact of continuous and intermittent thermal stress during early embryogenesis on hatchability, physiological body reaction, ovary weight, and follicle development of quails. A total of 540 eggs were divided into 3 equal groups (3 groups × 6 replicates × 30 eggs). In the first group (control), eggs were incubated at normal incubation conditions (37.5°C and 50–55% relative humidity) from day 0 till hatching. In the second group (continuous thermal stress [CTS]), eggs were daily exposed to 39.5°C and 50 to 55% during the early embryogenesis for 3 successive days (E4–E6) for 3 h (12:00–15:00). In the third group (intermittent thermal stress [ITS]), eggs were daily exposed to 39.5°C and 50 to 55% during the early embryogenesis for 90 min (12:00–13:30) then temperature was returned to 37.5°C for 60 min (13:30–14:30) after that the temperature was raised again for 39.5°C for 90 min (14:30–16:00) daily for 3 successive days (E4–E6). The findings showed that the highest relative water loss form egg (RWL/%) at 6 d of incubation was obtained in the CTS group (P ≤ 0.05). The hatchability rate was significantly (P ≤ 0.05) decreased in the thermal-treated groups compared with the control group. The body surface temperature and cloacal temperature in the CTS and ITS groups significantly (P ≤ 0.001) increased compared with the control group. Chick weight (g) at 5 wk old, total weight gain, daily weight gain were significantly lower (P ≤ 0.05) in the CTS group compared with the control group. Triiodothyronine (T3) hormone concentration and globulin level were significantly (P ≤ 0.05) lower in the CTS and ITS groups compared with the control. The ovarian follicle weights (first, second, third, fourth, and fifth) and the diameter of the large follicle (fifth follicle) were significantly (P ≤ 0.01) decreased by increasing incubation temperature. From these findings, it could be concluded that the hatchability and body weight at sexual maturity for quails produced from eggs exposed to CTS and IST were significantly decreased by 8 and 2.1% as well as 2.98 and 2.1%, respectively, compared with the control group.

MiRNA expression profile in chicken ovarian follicles throughout development and miRNA-mediated MMP expression

Accumulating evidence has demonstrated the role of microRNAs (miRs) in the avian ovary. In this study, high-throughput transcriptome analyses were employed to study the differential miR expression profiles in the chicken ovary, aiming to reveal miR-targeting matrix metalloproteinase (MMP) expression during follicular growth, maturation, and atresia. Using tissues of chicken ovarian follicles at key steps of development (slow growing - white, the most recently recruited - small yellow, and preovulatory - F2) and regression (the third postovulatory), 14 small RNA (sRNA) libraries were constructed. The 25 most highly expressed known miRs were identified along with eight significantly differentially expressed (DE) miRs (gga-miR-let-7d, gga-miR-31-3p, gga-miR-138-1-3p, gga-miR-1552-5p, gga-miR-92-3p, gga-miR-31-5p, gga-miR-202-3p, and gga-miR-6648-3p) which were further examined by quantitative real time-PCR (qRT-PCR) in white, yellowish, small yellow, and atretic follicles as well as in the granulosa and theca layer of yellow preovulatory F3–F1 follicles (n = 6 hens). These miRs were mainly associated with four pathways: inhibition of MMPs, axonal guidance signaling, HIF1α signaling, and GP6 signaling. Four predicted target genes (i.e. MMP-16, ADAM10, COL4A2, and COL4A5) were examined by qRT-PCR and negatively correlated with DE miRs. The identified candidate miR:mRNA target pairs include gga-miR-31-5p or gga-miR-92-3p:MMP-16, gga-miR-31-5p or gga-miR-92-3p:ADAM10, let-7d:COL4A2, and gga-miR-138-1-3p:COL4A5 are potentially associated with MMP modulation in the hen ovary, mostly in the granulosa and theca cells of the largest preovulatory follicles. These results provide a novel insight to the role of miRs in follicle development by identifying a miR target network that is putatively engaged in remodeling of the extracellular matrix during ovarian follicle development in chickens.

The effects of caffeine on ovarian tissue in rats.

Caffeine consumption increases during early adulthood, which has adverse effects on the reproductive system. This study aimed to assess the impact of embryonic caffeine exposure on rat ovary in adulthood. Female Wistar rats (240-270g) were divided into 5 groups (n = 7): experimental groups were exposed to 26, 45, 100, and 150mg/kg of caffeine via drinking water during pregnancy and the control group only received drinking water. The ovaries of the offspring were taken out on days 7, 14, 28, 60, 90, and 120 of postnatal development, and then, they were fixed in 10% formaldehyde solution. Ovarian follicles were studied using stereological methods, and data were analyzed using one-way ANOVA followed by the Tukey test in SPSS software. A value of p < 0.05 was considered significant. The body weight, the weight of the ovaries, the ovarian volume, and the number of primordial follicles decreased significantly (p < 0.05) in 45 and 100mg/kg, and (p < 0.001) in 150mg/kg caffeine-treated groups at all stages of postnatal development. Significant decreases were observed in the number of primary and secondary follicles in 45 and 100mg/kg (p < 0.05) and (p < 0.001) in 150mg/kg caffeine-treated groups on days 7, 14, 28, and 60 compared to the control group. The number of Graafian follicles also decreased significantly (p < 0.001) in 45, 100, and 150mg/kg caffeine-treated groups on days 14 and 28. Moreover, the mean volume of the oocyte in Graafian follicles reduced considerably in 45, 100, and 150mg/kg caffeine-treated groups compared to other groups (p < 0.05). The thickness of the zona pellucida (ZP) in the secondary follicles (p < 0.02) and Graafian follicles (p < 0.05) showed a significant reduction in 100 and 150mg/kg caffeine-treated groups on the 14th, 28th, and 60th days. In conclusion, high-dose caffeine consumption during gestation affects all stages of ovarian follicle development in rat offspring.

The Effect of Bone Marrow Transplantation on Oocyte-Granulosa Cell Interaction and Follicular Development of Cisplatin-Induced Ovarian Failure in Rat

Introduction: Chemotherapy has cytotoxic effect that induces follicular damage and abnormal folliculogenesis leads to ovarian failure. Two crucial growth factors in abnormal folliculogenesis, Growth Differentiation Factor-9 (GDF-9) and Kit-Ligand, will be disrupted and affect follicular development. In this study, we evaluate whether bone marrow transplantation (BMT) has a role on oocyte-granulosa cell interaction by analyzing GDF-9 and Kit-Ligand expressions and also follicular development by analyzing primordial, primary, secondary and graafian follicles of cisplatin-induced ovarian failure in rat. Material and method: Forty eight rats were divided into three groups: control, cisplatin and cisplatin + BMT. Ovarian failure was induced by administration of intraperitoneal cisplatin 5 mg/kg body weight for 1 week. BMT 2 x107 cells were injected through rat tail vein after cisplatin administration. Bone marrow was isolated from rat femur and characterized by CD44(+), CD45(-), CD105(+). Immunohistochemistry examinations for ovarian GDF-9, Kit-Ligand and follicle development evaluation were performed after 2 weeks of BMT injection. Results: The expressions of Kit-ligand among three groups by ANOVA were significant different (p=0.00), whereas by Post Hoc: cisplatin group lower than control group (p=0.00); cisplatin + BMT group higher than cisplatin group (p=0.00); and no significant different between control group and cisplatin + BMT group (p=0.955). The expressions of GDF-9 by Kruskal Wallis showed significant different (p=0.00) among three groups whereas cisplatin + BMT group higher than cisplatin group and control group. In cisplatin + BMT group the number of primordial, primary, secondary and graafian follicles were higher than those in cisplatin group; but were lower than those in control group (p=0.000). Positive Paul Kart Horan (PKH) labeling was seen in cisplatin + BMT group only. Conclusion: In cisplatin-induced ovarian failure in rat, bone marrow transplantation may improve oocytegranulosa cell interaction and follicular development.

Anti-M\xfcllerian hormone levels and risk of type 2 diabetes in women

Aims/hypothesisGiven its role in ovarian follicle development, circulating anti-Müllerian hormone (AMH) is considered to be a marker of reproductive ageing. Although accelerated reproductive ageing has been associated with a higher risk of type 2 diabetes, research on the relationship between AMH and type 2 diabetes risk is scarce. Therefore, we aimed to investigate whether age-specific AMH levels and age-related AMH trajectories are associated with type 2 diabetes risk in women.MethodsWe measured AMH in repeated plasma samples from 3293 female participants (12,460 samples in total), aged 20–59 years at recruitment, from the Doetinchem Cohort Study, a longitudinal study with follow-up visits every 5 years. We calculated age-specific AMH tertiles at baseline to account for the strong AMH–age correlation. Cox proportional hazards models adjusted for confounders were used to assess the association between baseline age-specific AMH tertiles and incident type 2 diabetes. We applied linear mixed models to compare age-related AMH trajectories for women who developed type 2 diabetes with trajectories for women who did not develop diabetes.ResultsDuring a median follow-up of 20 years, 163 women developed type 2 diabetes. Lower baseline age-specific AMH levels were associated with a higher type 2 diabetes risk (HRT2vsT3 1.24 [95% CI 0.81, 1.92]; HRT1vsT3 1.62 [95% CI 1.06, 2.48]; ptrend = 0.02). These findings seem to be supported by predicted AMH trajectories, which suggested that plasma AMH levels were lower at younger ages in women who developed type 2 diabetes compared with women who did not. The trajectories also suggested that AMH levels declined at a slower rate in women who developed type 2 diabetes, although differences in trajectories were not statistically significant.Conclusions/interpretationWe observed that lower age-specific AMH levels were associated with a higher risk of type 2 diabetes in women. Longitudinal analyses did not show clear evidence of differing AMH trajectories between women who developed type 2 diabetes compared with women who did not, possibly because these analyses were underpowered. Further research is needed to investigate whether AMH is part of the biological mechanism explaining the association between reproductive ageing and type 2 diabetes.Graphical abstract

Corrigendum to “Determine the Role of FSH Receptor Binding Inhibitor in Regulating Ovarian Follicles Development and Expression of FSHR and ERα in Mice”

[This corrects the article DOI: 10.1155/2018/5032875.].

Long-Term Advantages of Ovarian Reserve Maintenance and Follicle Development Using Adipose Tissue-Derived Stem Cells in Ovarian Tissue Transplantation.

(1) Background: Ovarian tissue transplantation with adipose tissue-derived stem cells (ASCs) has been shown to enhance graft vascularization and increase follicle survival after a short interval of 7 days. The aim of the present study was to investigate their long-term effects on primordial follicle pool maintenance and follicle development. (2) Methods: A total of 14 severe combined immunodeficient (SCID) mice were grafted with frozen-thawed human ovarian tissue with or without ASCs. Blood was taken monthly in order to quantify the anti-Müllerian hormone (AMH) and estradiol. After 6 months, all the grafts were retrieved and sent for histology and immunolabeling (AMH, AMH receptor II, estrogen receptors α and β, and c-kit/kit ligand). (3) Results: A significant upturn was observed in AMH and estradiol plasma levels 4 months after transplantation in both grafted groups. The primordial follicle pool was better preserved in the ASC group (41.86 ± 28.35) than in the standard transplantation group (9.65 ± 17.6, p < 0.05) compared to non-grafted controls (124.7 ± 140). (4) Conclusions: The use of ASCs prior to ovarian tissue transplantation yielded a larger primordial follicle pool and more physiological follicle distribution after long-term grafting. These findings suggested that ASC use might extend the ovarian tissue lifespan.