Single cell transcriptome atlas of microenvironment in preovulation follicle
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE189960 | GEO | 2022/06/22
REPOSITORIES: GEO
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Project description:Cows in Negative Energy Balance (NEB) may preferentially divert nutrients away from reproduction, thereby experiencing a period of anovulatory anestrus, delayed ovulation of large follicles and a condition of impaired fertility. To better understand the changes occurring in these large follicles as a function of time post-partum granulosa cells of preovulatory follicles have been collected at different times: 30, 60, 90 and 120 days after calving . An analysis of the transcriptome was performed using a global bovine oligo-array microarray to map the differences in genes expression and cellular functions that occur in the follicular microenvironment during the progressive recovery from NEB condition in dairy cow
2013-12-31 | GSE50854 | GEO
Project description:Cows in Negative Energy Balance (NEB) may preferentially divert nutrients away from reproduction, thereby experiencing a period of anovulatory anestrus, delayed ovulation of large follicles and a condition of impaired fertility. To better understand the changes occurring in these large follicles as a function of time post-partum granulosa cells of preovulatory follicles have been collected at different times: 30, 60, 90 and 120 days after calving . An analysis of the transcriptome was performed using a global bovine oligo-array microarray to map the differences in genes expression and cellular functions that occur in the follicular microenvironment during the progressive recovery from NEB condition in dairy cow Four time points experiment: 30, 60, 90 and 120 days. Granulosa cells from the 30, 60 and 90 days compare to the 120 days (reference). Biological replicates: 3 from each time point. One replicate per array.
2013-12-31 | E-GEOD-50854 | biostudies-arrayexpress
Project description:Gonadotropin-releasing hormone (GnRH) might combine with other reproductive hormones to affect follicular development by regulation of the cell cycle in granulosa cells (GCs).To investigate the function of GnRH in GCs, we examined the effects of a GnRH agonist (alarelin acetate) and an antagonist (cetrorelix acetate) on the proliferation of primary goose (Anser cygnoides) ovarian GCs by colorimetry and 5-ethynyl-2¢-deoxyuridine (EdU) cell proliferation assays. Differently expressed genes (DEGs) affected by these GnRH analogues in goose GCs were detected by RNA-seq and the results were verified using quantitative reverse transcription polymerase chain reaction (RT–qPCR) and western blotting. The optical densities in the colorimetry assays and the S phase cell cycle ratio of GCs in the EdU cell proliferation assays gradually increased and decreased with increasing concentrations in the alarelin-treated and cetrorelix-treated cells in dose-dependent manners, respectively. Statistically significances were achieved at concentrations of 10−5 mol/L alarelin and 1 mg/L cetrorelix acetate RNAseq analysis showed that 134 DEGs and 226 DEGs were detected following GC treatment with 10−5M GnRH alarelin (76 downregulated and 58 upregulated) and 1 mg/L cetrorelix (90 downregulated and 136 upregulated). Many of these DEGs are involved in the actions of genes encoding G protein-coupled hormone receptors, such as estrogen receptor 2 (ESR2), oxytocin receptor (OXTR), and prolactin receptor (PRLR), which are essential for folliculogenesis. The mRNA and protein expression levels of ESR2 in GCs were decreased and increased by the GnRH agonist and antagonist, respectively. Interestingly, ESR2 was increasingly expressed in pre-hierarchical follicles, most strongly expressed in GCs of F5 stage follicles and reduced in preovulatory follicles, which implicates a potential role in follicle selection. Thus, GnRH might act as an autocrine or paracrine factor to regulate GC proliferation by interacting with some key steroid hormone G protein-coupled receptors genes participating in follicular development.
2019-10-16 | GSE133337 | GEO
Project description:The Objective of this study was to establish global gene expression profiles associated with different stages of dominant follicle development in the horse. This was done by collecting ovaries at different stages of an ovulatory follicular wave, when the dominant follicle reached 22 mm (early dominant, ED), 33 mm (late dominant, LD) and 34 h after an i.v. injection of crude equine gonadotropins (CEG; 15 mg i.v.) when the largest follicle reached > 33 mm (preovulatory stage, PO). RNA was then separately collected from granulosa cells and theca-rich follicular wall fractions and was hybridized to the Agilent Horse Gene Expression Microarray. Gene expression was compared between ED and LD and between LD and PO stages within each of granulosa cells and theca walls. Significantly larger numbers of genes were differentially expressed in granulosa cells than in theca walls throughout development, and between LD and PO follicles than between ED and LF follicles. The most salient features were the downregulation of cell cycle genes in granulosa cells during both the ED-LD and LD-PO transitions and the upregulation of genes associated with inflammation, immunity, extracellular matrix remodelling and protection aganints toxic insult in both GCs and TCs, particularly during the LD-PO transition.
2014-03-15 | GSE52109 | GEO
Project description:In the ovary, proliferation and differentiation of granulosa cells (GCs) drive the growth of follicles. This is, in part, dependent on gonadotropins. Our immunohistochemical studies provided hints of mitochondrial biogenesis and intracellular redistribution in GCs of growing follicles. A cellular model, human KGN cells (granulosa cell tumor cells, derived from growing follicles) was used to study aspects of mitochondrial dynamics. To elevate cAMP and thereby mimic gonadotropin actions, forskolin (FSK) was used, which increased KGN cell size and mitochondrial DNA within 24 h. MitoTracker experiments and ultrastructural 3D reconstruction revealed that FSK treatment induced the formation of elaborate mitochondrial networks. H89, a protein kinase A (PKA) inhibitor, reduced network formation. A proteomic analysis indicated that FSK among others elevated levels of regulators of the cytoskeleton and the steroidogenic enzyme CYP11A1, located in mitochondria, was more than 3-fold increased, implying that cAMP/PKA-associated structural changes go in parallel with the acquisition of steroidogenic competence of mitochondria in KGN cells. In summary, in situ observations showed increases in mitochondria and suggested intracellular trafficking in GCs during follicular growth and indicated that they may partially be under the control of gonadotropins/cAMP. In line with this, elevation of cAMP in KGN profoundly affected mitochondria dynamics in a PKA-dependent manner and implicated cytoskeletal changes.
2022-10-14 | PXD032160 | Pride
Project description:Follicles of polycystic ovaries (PCO) often become arrested in early antral stages at around 3 to 11 mm in diameter. The condition disturbs dominant follicle selection and may result in altered ovulation and anovulation. During the growth and development of human follicles, the follicular fluid (FF) constitutes the avascular microenvironment in which the oocyte develops and acts as a vehicle for hormone signaling between cues from circulation and follicular cells. Previous proteomics studies performed on FF from women with polycystic ovarian syndrome (PCOS) have revealed information on the protein changes associated with the pathophysiology of this disorder. However, these studies have been conducted on FF samples obtained in connection with oocyte pick-up during ovarian stimulation right at the time of ovulation and are limited to follicular conditions during the follicular phase of the menstrual cycle. This study aimed to detect proteomic alterations in FF from human small antral follicles (hSAF) obtained from women with PCO as compared to normal women.
2022-04-07 | PXD031485 | Pride
Project description:The Objective of this study was to establish global gene expression profiles associated with different stages of dominant follicle development in the horse. This was done by collecting ovaries at different stages of an ovulatory follicular wave, when the dominant follicle reached 22 mm (early dominant, ED), 33 mm (late dominant, LD) and 34 h after an i.v. injection of crude equine gonadotropins (CEG; 15 mg i.v.) when the largest follicle reached > 33 mm (preovulatory stage, PO). RNA was then separately collected from granulosa cells and theca-rich follicular wall fractions and was hybridized to the Agilent Horse Gene Expression Microarray. Gene expression was compared between ED and LD and between LD and PO stages within each of granulosa cells and theca walls. Significantly larger numbers of genes were differentially expressed in granulosa cells than in theca walls throughout development, and between LD and PO follicles than between ED and LF follicles. The most salient features were the downregulation of cell cycle genes in granulosa cells during both the ED-LD and LD-PO transitions and the upregulation of genes associated with inflammation, immunity, extracellular matrix remodelling and protection aganints toxic insult in both GCs and TCs, particularly during the LD-PO transition. Five to nine biological replicates per cell type and developmental stage (total of 34 samples) were used in a single dye experiment. Samples were distributed among slides so that each experimental groupeach was represented at least once in each slide. For each gene, mean normalized intensities (n= 5 biological replicates/group) were compared between follicle stages (ED-LD and LD-PO) within each cell type (GC and TC).
2014-03-15 | E-GEOD-52109 | biostudies-arrayexpress
Project description:Human primary granulosa cells (GCs) derived from women, undergoing oocyte retrieval, can be cultured and are a cellular model for the study of human ovarian functions. In vitro they change rapidly, resembling initially cells of the preovulatory follicle and then cells of the corpus luteum. They are derived from individual patients and their different medical history, lifestyle and age lead to heterogeneity. Thus, cells can rarely be ideally matched for cellular experiments or, if available, only in small quantities. We reasoned that cryopreservation of human GCs may be helpful. Previous studies indicated feasibility of such an approach, but low survival of GCs was reported and consequences on GC functionality were only partially evaluated. We tested a slow freezing protocol (employing FCS and DMSO) of GCs upon isolation from follicular fluid. We compared cryopreserved and subsequently thawed cells with fresh, not cryopreserved ones, from the same patients. About 80 % of human GCs survived freezing/thawing. Neither morphology, nor levels of cell-cell contact, mitochondrial and steroidogenic genes were different between the two groups in cells cultured for 1-5 days. A proteomic analysis revealed no statistical significance in the abundance of a total of 5962 proteins. Both groups produced comparable basal levels of progesterone and responded similarly to hCG with elevation of progesterone. Taken together, we describe a rapid and readily available method for the cryopreservation of human GCs. We anticipate that it will allow future large-scale experiments and may thereby improve cellular studies with human ovarian cells.
2021-07-28 | PXD025632 | Pride
Project description:Cellular mechanisms that contribute to low estradiol concentrations produced by the preovulatory ovarian follicle in cattle with a compromised metabolic status (such as lactatino) are largely unknown. To gain insight into the main metabolic mechanisms affecting preovulatory follicle function RNAseq profiling was conducted on non-lactating Holstein-Friesian heifers (n=16) and lactating Holstein-Friesian cows (n=17) at three stages of preovulatory follicle development: A) newly selected dominant follicle in the luteal phase (Selection); B) follicular phase before the LH surge (Differentiation) and C) pre-ovulatory phase after the LH surge (Luteinization). Based on a combination of RNA sequencing, ingenuity pathway analysis and Q-RT-PCR validation several important molecular markers involved in steroid biosynthesis, such as the expression of steroidogenic acute regulatory protein (STAR) within developing dominant follicles, were identified to be affected (downregulated) by the catabolic state. We propose that the adverse metabolic environment caused by lactation decreases preovulatory follicle function by affecting cholesterol transport into the mitochondria to initiate steroidogenesis.
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Project description:Human ovarian folliculogenesis is an highly regulated and complex process. Characterization of follicular cell signatures during this dynamic process is important to understand follicle fate (to grow, become dominant or undergo atresia). The transcriptional signature of human oocytes and granulosa cells (GC) in early-growing and ovulatory follicles have been previously described, however that of oocytes with surrounding GCs in small antral follicles have not been studied yet. Here, we have generated a unique dataset of single-cell transcriptomics (SmartSeq2) consisting of the oocyte with surrounding GCs from several individual (non-dominant) small antral follicles isolated from adult human ovaries. We have identified two main types of (healthy) follicles, with a distinct oocyte and GC signature. Using the CellphoneDB algorithm, we then investigated the bi-directional ligand-receptor interactions regarding the TGFβ/BMP, WNT, NOTCH, and receptor tyrosine kinases (RTK) signaling pathways between oocyte and GCs within each antral follicle type. Our work not only revealed the diversity of small antral follicles but also contributes to fill the gap in mapping the molecular landscape of human folliculogenesis and oogenesis.
2021-10-26 | GSE186504 | GEO