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To compare the transcriptome expression of cumulus granulosa cells between PCOS patients with insulin resistance and women undergoing assisted reproduction for fallopian tube reasons

ABSTRACT: Purpose polycystic ovary syndrome (PCOS) is the most common endocrine and metabolic disorder in women of childbearing age. It is characterized by follicular dysplasia and hyperandrogenemia. Insulin resistance plays an important role in the development of PCOS.However, the mechanism of regulation and interaction between insulin resistance and follicle development remains unclear.The aim of this study was to identify differential genes associated with follicle development in cumulus granulosa cells in PCOS patients with insulin resistance compared with controls. Methods In this study, RNA-seq, bioinformatics analysis and reverse transcription qPCR (RT-qPCR) were used to analyze the gene expression in cumulus granulosa cells.A total of seven cDNA libraries were constructed from three PCOS samples and four control samples.3 μg of RNA was used as starting material for each sample.Ribosomal RNA was removed using the Epicentre rigo-zero ™ Gold Kit (Epicentre, Illumina, Madison, WI, USA). Results Compared with the control group, there were 2233 DEGs in the PCOS group, including 1158 up-regulated genes and 1075 down-regulated genes. Some different expression patterns were observed in the two groups.The next series of experiments verified the effect of some genes on oocyte development. Conclusion Our study showed that the differential expression of transcriptome in cumulus granulosa cells could affect the growth, development and maturation of oocytes.Meanwhile, the results of RT-PCR and Elisa showed that the expression of renin angiotensin system was changed in PCOS patients with insulin resistance, and these changes could affect oocyte maturation and excretion through cumulus granulosa cells.

ORGANISM(S): Homo sapiens

PROVIDER: GSE216609 | GEO | 2023/10/01

REPOSITORIES: GEO

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Project description:In mammals, the capacity of the female germ cell, the oocyte, to develop into embryo is acquired throughout meiotic maturation. Immature oocyte cannot be fertilized while mature oocyte is apt to accept spermatozoa and to develop an embryo. In a follicle, the oocyte is surrounded by mural granulosa cells (GC) and is physically and metabolically coupled with specialized granulosa cumulus cells (CC) which play an important role in oocyte maturation and fertilization. Factors expressing in GC and CC during maturation may reflect the oocyte quality, i.e. its capacity to be fertilized and assure early embryo development. However, the modifications of the content and the amount of peptide/proteins in the oocyte and the surrounding CC during oocyte maturation are mostly unknown and so there is not an accurate way of evaluating/monitoring how different in vitro maturation (IVM) protocols being in use in assisted reproduction technologies, can affect the process. In this context, Intact Cell MALDI-TOF Mass Spectrometry (ICM-MS) was applied to bovine follicular cells (bovine single oocytes, cumulus cells and granulosa cells) in order to characterize proteomic changes that occur in the follicle during female gamete development. In order to characterize finely endogenous molecular species observed on ICM-MS profiles and to identify markers of interest with their post-translational modifications, we carried out top-down proteomic on the different follicular cells from oocyte, oocyte-cumulus complexes, cumulus cells and granulosa cells protein extracts. Prior to top-down MS using a dual linear ion trap Fourier Transform Mass Spectrometer LTQ Orbitrap Velos, depending on the amount of available biological material, we employed three analytic strategies as a direct infusion, a mono-dimensional liquid chromatography (µLC-1D-MS/MS) and an off-line multi-dimensional liquid chromatography combining four fractionations (based on reverse phase or gel filtration LC) to µLC-MS/MS. Here, we deposited our dataset from µLC-1D-MS/MS (analyses of oocytes, oocyte-cumulus complexes, cumulus cells and granulosa cells protein extracts) and MDLC-MS/MS (analyses of granulosa cells protein extract).

Project description:Direct exposure to physiologically-relevant elevated temperatures during the first half of in vitro maturation heightens cumulus-derived progesterone production, suppresses matrix metallopeptidase 9 secretion, and alters cumulus transcriptome. We hypothesized that heat-induced perturbations in cumulus cells enveloping maturing oocyte may extend to the mural granulosa of the periovulatory follicle in the heat-stressed cow to subsequently follicular fluid proteome. Lactating Holsteins were pharmacologically stimulated to produce a dominant follicle then given GnRH to induce an LH surge. Following GnRH administration, cows were maintained at ~67 temperature humidity index (THI; thermoneutral conditions) or exposed to conditions simulating an acute heat stress event (71 to 86 THI; heat stress for ~12 h). Fluid was aspirated from periovulatory follicle ~16 h after GnRH. Follicular fluid proteome from thermoneutral (n = 5) and hyperthermic (n = 5) cows was evaluated by quantitative tandem mass spectrometry (nano LC-MS/MS). We identified 35 differentially-abundant proteins. Functional annotation revealed numerous immune-related proteins. Subsequent efforts revealed an increase in levels of the proinflammatory mediator bradykinin in follicular fluid (P = 0.0456) but not in serum (P = 0.9319) of hyperthermic cows. Intrafollicular increases in transferrin (negative acute phase protein) in hyperthermic cows (P = 0.0181) coincided with a tendency for levels to be increased in the circulation (P = 0.0683). Nine out of 15 cytokines evaluated were detected in follicular fluid. Heat stress increased intrafollicular IL6 levels (P = 0.0160). Whether hyperthermia-induced changes in the heat-stressed cow’s follicular fluid milieu reflect changes in mural granulosa, cumulus, other cell types secretions, and/or transudative changes from circulation remains unclear. Regardless of origin, heat stress/hyperthermia related changes in the follicular fluid milieu may have an impact on components important for ovulation and competence of the cumulus-oocyte complex contained within the periovulatory follicle

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To compare the transcriptome expression of cumulus granulosa cells between PCOS patients with insulin resistance and women undergoing assisted reproduction for fallopian tube reasons

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