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The Transcriptome Landscape change of Human Granulosa Cells under gonadotropins Revealed by Small bulk Cell RNA Sequencing

ABSTRACT: STUDY QUESTION: Can superovulation dose of gonadotropins (Gns) affect the transcriptome of granulosa cells and follicular fluid (FF) hormone levels? SUMMARY ANSWER: Cell cycle- and meiosis-associated genes were down-regulated by Gns stimulation, whereas immune- and cytokine-associated genes were up-regulated. Hormone levels were also affected by Gns stimulation. WHAT IS KNOWN ALREADY: Superovulation treatment had adverse effects on maturation, spindle function, and chromosomal aneuploidy in oocytes. STUDY DESIGN, SIZE, DURATION: One leading pre-ovulatory follicle per subject was used from three natural-cycle and four Gns-stimulated patients. PARTICIPANTS/MATERIALS, SETTING, METHODS: Granulosa cells and FF were collected from the same leading follicle from each patient. RNA was extracted from granulosa cells and subjected to deep sequencing and analysis. Follicle-stimulating hormone (FSH), estradiol (E2), androstenedione (AND), testosterone (T), luteinizing hormone (LH), and progesterone (P) levels in FF were measured by immunoassays. Student s t test was used for statistical analysis. MAIN RESULTS AND ROLE OF CHANCE: A total of 715 genes were up-regulated, and 287 genes were down-regulated, in the Gns-stimulated group relative to the Control group. Gene Ontology analysis revealed that the down-regulated genes were enriched in cell cycle and meiosis pathways, primarily those associated with follicle or oocyte maturation and quality. On the other hand, the up-regulated genes were enriched in functions related to immunity and cytokine Ccytokine receptor interactions. In stimulated vs. natural-cycle follicles, E2 and LH levels were significantly reduced (P < 0.001).

ORGANISM(S): Homo sapiens

PROVIDER: GSE124177 | GEO | 2021/12/20

REPOSITORIES: GEO

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Project description:The molecular mechanisms that regulate the pivotal transformation processes observed in the follicular wall following the pre-ovulatory LH-surge, are still not established, particularly for cells of the thecal layer. To elucidate thecal and granulosa cell type-specific biological functions and signaling pathways, large dominant bovine follicles were collected before and 21 hrs after an exogenous GnRH induced LH surge. Because LH receptor density varies within the granulosa cell populations, antral granulosa (aGC; those aspirated by follicular puncture) and membrane associated granulosa (mGC; those scraped from the follicular wall) were compared to thecal cell expression profiles determined by mRNA microarrays. Thecal cell gene expression was less affected in the peri-ovulatory follicle when compared to granulosa cells, as evidenced by only 2% versus 25% of the ~11,000 genes expressed changing in response to the LH surge, respectively. The majority of the 203 LH-regulated thecal genes were also LH regulated in granulosa cells, leaving a total of 58 genes as LH-regulated theca cell specific genes. Most of the 58 genes (i.e., 74%) thecal specific genes including several known thecal markers (CYP17A1, NR5A1) were downregulated, while most genes identified are new to theca. Many of the newly identified upregulated thecal genes (e.g., PTX3, RND3, PPP4R4) were also upregulated in granulosa. Minimal expression differences were observed between aGC and mGC, however, transcripts encoding extracellular proteins (NID2) and matrix modulators (ADAMTS1, SASH1) predominated these differences. We also identified large numbers of unknown LH-regulated granulosa cell genes and discuss their putative roles in ovarian function. The single dominant ovarian follicle was collected from each cow before the LH surge or 22 hours after GnRH (used to induce LH surge). RNA was extracted from three independent cells within each follicle and there were hybridized on Affymetrix microarrays.

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The Transcriptome Landscape change of Human Granulosa Cells under gonadotropins Revealed by Small bulk Cell RNA Sequencing

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