Vitrification preserves follicular transcriptomic dynamics during ex vivo ovulation
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA943225 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR23802600_1.fastq.gz | Fastqsanger.gz | |||
| SRR23802600_2.fastq.gz | Fastqsanger.gz | |||
| SRR23802601_1.fastq.gz | Fastqsanger.gz | |||
| SRR23802601_2.fastq.gz | Fastqsanger.gz | |||
| SRR23802602_1.fastq.gz | Fastqsanger.gz |
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Project description:Vitrification is a method for long-term biological sample cryopreservation without causing intra- and extra-cellular ice formation. We recently established a novel closed vitrification system to cryopreserve mouse ovarian follicles. Using the 3D alginate hydrogel encapsulated in vitro follicle growth (eIVFG) method, we have demonstrated that compared to freshly-harvested follicles, vitrified follicles have normal follicle and oocyte reproductive outcomes. Our recent study further demonstrated the faithful preservation of molecular signatures of follicle-stimulating hormone (FSH)-stimulated follicle maturation in vitrified follicles. However, it is unknown whether ovulation, another crucial gonadotropin-dependent follicular event, and involved ovulatory gene regulatory pathways are well conserved in vitrified follicles. Fresh and vitrified follicles grown for 8 days by eIVFG were collected at 0, 1, 4 and 8-hour post human chorionic gonadotropin (hCG) treatment for the single-follicle RNA sequencing. Principal component analysis (PCA) and Pearson’s correlation analysis revealed that vitrified follicles have similar transcriptomic profiles to fresh follicles. Furthermore, the expression of several genes essential for ovulation were comparable between vitrified and fresh follicles. In summary, these results demonstrate that our closed vitrification system preserves follicular transcriptomic dynamics during ex vivo ovulation. Together with our previous findings that vitrification preserves FSH-stimulated follicle maturation, the integration of vitrification of immature follicles and eIVFG has a great potential to serve as an additional fertilization preservation approach for young cancer patients and endangerers species. Moreover, follicle vitrification enables a high-content ovarian follicle biobank, which can greatly improve the throughput of eIVFG for studying ovulation biology, anovulatory disease, toxicology, and novel contraceptive drug development targeting ovulation.
2023-09-20 | GSE227091 | GEO
Project description:Vitrification is a method for long-term biological sample cryopreservation that transforms cells into a glass-like state by cooling without causing intra- and extra-cellular ice formation, which is a major driver of cell cryoinjury. Using an alginate hydrogel encapsulated in vitro follicle growth (eIVFG) system, we have recently demonstrated that compared to freshly-harvested follicles, vitrified follicles have normal follicle and oocyte reproductive outcomes as well as comparable expression levels of several genes that are essential for gonadotropin-dependent folliculogenesis and oogenesis. However, it is unknown whether vitrification preserves the molecular signatures of folliculogenesis at the whole transcriptomic level, which is the primary research focus in this study. Multilayered secondary follicles with a diameter of 130 -160 µm were mechanically isolated from 16-day-old CD-1 female mice and vitrified using the closed vitrification method. After a 2-week storage in liquid nitrogen, vitrified follicles were warmed, and together with freshly-harvested follicles, cultured for 8 days using the eIVFG method. Next, mature antral follicles from day 8 were collected for single-follicle RNA sequencing using the SMART-seq RNA-seq method. Principal component analysis (PCA) and Pearson’s correlation analysis revealed that vitrified follicles have similar transcriptomic profiles to fresh follicles. Using the criterion of fold change >2 or <0.5 and a false discovery rate (FDR) adjusted p-value < 0.05, there were 35 differentially expressed genes out of a total of 11,806 detected genes between vitrified and fresh follicles, among which 17 were up-regulated and 18 were down-regulated in vitrified follicles. To the best of our knowledge, none of those differentially expressed genes identified between vitrified and fresh follicles have been shown to be critical to folliculogenesis and oogenesis. Meanwhile, gene ontology (GO) and KEGG pathway analysis over these 35 genes using the functional annotation tool DAVID revealed that no GO terms or signaling pathways were significantly enriched. Furthermore, the expression of several genes essential for the gonadotropin-dependent folliculogenesis and oogenesis were comparable between vitrified and fresh follicles, including granulosa cell-specific genes (Cyp19a1, Amh, Inha, Inhba, Inhbb, and Fshr), theca cell-specific genes (Cyp17a1, Col1a2, and Col3a1), and oocyte-specific genes (Gdf9, Bmp15, Zp1, Zp2, and Zp3). Equally important, Lhcgr, the gene that is primarily expressed in theca cells of preantral follicles and then greatly induced in mural granulosa cells in antral follicles to prepare for the luteinizing hormone (LH) surge and ovulation, also had comparable mRNA expression levels between vitrified and fresh follicles. Taken together, these results demonstrate that vitrification preserves the follicular cell transcriptome and molecular signatures of gonadotropin-dependent folliculogenesis in the eIVFG system. In conclusion, our study re-evaluates individual follicle cryopreservation by using the closed vitrification method in a more in-depth perspective and proves that vitrification preserves the follicular cell transcriptome in the eIVFG system, thus providing a robust model for fertility preservation, conservation of endangered species, and also establishing a high-content ovarian follicle biobank for studying ovarian biology and female reproductive toxicology.
2021-12-29 | GSE184691 | GEO
Project description:Vitrification preserves murine ovarian follicular cell transcriptome in a 3D encapsulated in vitro follicle growth system
| PRJNA765674 | ENA
Project description:To investigate the transcriptomic profile of in vivo and ex vivo ovulation systems
2024-07-08 | GSE255274 | GEO
Project description:The composition of follicular fluid reflects crucial paracrine signalling between granulosa- and theca cells and the oocyte. Dynamic changes in the protein composition of human follicular fluid across multiple time points in the period where final meiotic maturation and release of the oocyte take place are previously undescribed. Twenty-five women undergoing IVF- or ICSI treatment in a standard antagonist protocol with agonist ovulation trigger were included in this prospective cohort study. From each patient one follicle was aspirated by transvaginal ultrasound guided puncture at one of five specific time points either before ovulation induction (T=0) or 12-, 17-, 32- or 36 hours after ovulation induction (five patients/time point). Proteomics was carried out by liquid chromatography-mass spectrometry. In total, 400 proteins were identified (FDR<0.05) and 30 were significantly regulated across time points (one-way ANOVA, adjusted p<0.05). Compared to compiled human plasma proteome sets, 47 proteins were unique to follicular fluid. Enriched biological functions among differentially expressed proteins included immune functions, wound healing and functions related to blood coagulation (FDR<0.02). The most profound changes occurred shortly after ovulation induction. We confirmed parallel protein expression to known granulosa cell mRNA changes and described many hitherto unknown protein expression profiles during ovulation. Thus, the study endorsed important biological functions of some proteins and suggested additional proteins, which may be crucial to intrafollicular signalling and oocyte competence that should be further investigated.
2019-07-29 | PXD012561 | Pride
Project description:Ex vivo imaging reveals the spatiotemporal control of ovulation
| PRJNA1074329 | ENA
Project description:Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disease which leads to serious impairment of reproductive health in women of child-bearing age. Anovulation or oligo-ovulation is a common clinical manifestation of PCOS patients. The disturbance of ovarian immune microenvironment contributes to the disorders of follicle development and ovulation, and the mechanism remains unclear. Here we demonstrated the protective effect of immune factor interleukin-22 (IL-22) on PCOS follicle development and ovulation. Follicular IL-22 levels were significantly lower in PCOS patients than in the control group and were positively correlated with the oocyte fertilization rate and the high-quality embryo rate. Additionally, IL-22 evidently improved follicle development in vitro and promoted ovulation-related gene expression, which was disrupted by the depletion of interleukin-22 receptor 1 (IL-22R1) or inhibition of STAT3 in granulosa cells, indicating that IL-22 acted through IL-22R1 and STAT3 signalling pathway to promote follicle development and ovulation in PCOS. In summary, this study elucidated the vital role of ovarian immune microenvironment in follicle development and ovulation, the application of IL-22 may provide new insights into the treatment of PCOS patients.
2024-10-09 | GSE266340 | GEO
Project description:Vitrification is replacing slow freezing as the most popular method for human embryo cryopreservation in clinics world-wide. Several studies demonstrated that cryopreservation alters gene expression of mammalian embryos, but none of them analysed what happen with those embryos that get implanted and follow with the gestation. The aim of this study was to evaluate the effect of vitrification technique on rabbit embryonic and fetal development by performing a transcriptomic analysis of 6 day old embryos and 14 days old fetal placentas. Effect of vitrification on late blastocyst and fetal placenta transcriptome. Four indepent replicates were performed for each condition (control and vitrified) and for both tissues (embryo and fetal placenta).
2013-09-26 | E-GEOD-51208 | biostudies-arrayexpress
Project description:Ovulation refers to the process when the ovarian surface-facing wall of a preovulatory follicle ruptures and releases the cumulus oocyte complex (COC) into the oviduct or fallopian tube in response to hormonal cues. In parallel, the unruptured wall of the follicle within the ovary transitions to becoming a progesterone-producing corpus luteum. Ovulation is essential for fertilization and eventual pregnancy. Disruption of ovulation, whether purposefully through contraceptive intervention or idiopathically in cases of anovulatory infertility, has translational implications for human health. Importantly, key processes of ovulation, including follicle rupture and luteinization, are recapitulated in models of ex vivo ovulation despite the absence of an intact hypothalamic-pituitary-gonadal axis and intra-ovarian cues. In our study, we used an ex vivo ovulation model to identify functional and molecular differences between distinct regions of the follicle wall, which we refer to as the ruptured and unruptured sides. We observed that the unruptured side of the follicle wall exhibits hallmarks of luteinization after ovulation while the ruptured side exhibits signs of cell death. RNA-sequencing of these specific follicle regions revealed 2,099 differentially expressed genes between follicle sides without hCG exposure and 1,673 between follicle sides 12 hours post-hCG, which were further validated in vivo. We found enriched pathways that recapitulate known ovulation biology, including oxidative stress on the ruptured side and angiogenesis on the unruptured side. We also identified previously unappreciated pathways that may play an important role in ovulation, such as amino acid transport and Jag-Notch signaling on the ruptured side, as well as metal ion processing and IL-11 signaling on the unruptured side. Ultimately our studies demonstrate that our ex vivo model recapitulates known in vivo ovarian biology, identifies pathways that may be novel regulators of ovulation and luteinization, and may have future translational applications for the study of ovulatory disorders and the development of novel non-hormonal contraceptives.
2024-10-18 | GSE266449 | GEO
Project description:Camelidae are considered to be inducing ovulators. Semen from these male animals has been identified to be rich in ovulation-inducing factors-NGF (β-nerve growth factor). Ovulation process is a vital and tightly regulated from multiple factors. Understanding the biological mechanisms underlying ovulation processes follicular development and hormone secretion will benefit the evidence the potential molecule mechanism of inducing ovulation. However, very little is known about the roles of these molecules in ovary. To identify candidate biomarkers of inducing ovulators.
2019-11-26 | PXD016451 |