Unknown,Transcriptomics,Genomics,Proteomics

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Functional germ line stem cells do not exist in adult mammalian ovaries

ABSTRACT: Single-cell RNA-seq of DDX4 FACS sorted cells from ovarian cortical tissue. Regarding singe cell quality, genes were filtered based on a RPKM greater than or equal to 1 in at least one cell, where the RPKM cutoff was determined from histograms of number of expressed transcripts versus RPKM, and it was also sensible with respect to the number of molecules per RPKM estimated from the ERCC spike-in. Three clear outliers were observed in a principal component analysis and therefore removed from further analysis.

INSTRUMENT(S): Illumina HiSeq 2000

ORGANISM(S): Homo sapiens

SUBMITTER: Outi Hovatta

PROVIDER: E-MTAB-2983 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The oocytes found within the primordial follicles of mammalian ovaries remain quiescent for months to years until they receive the appropriate signals to undergo the primordial to primary follicle transition and initiate folliculogenesis. The molecular mechanisms and extracellular signaling factors that regulate this process remain to be fully elucidated. The current study investigates the mechanisms utilized by anti-MÃ¼llerian hormone (AMH; i.e. MÃ¼llerian inhibitory substance) to inhibit the primordial to primary follicle transition. Ovaries from 4-day-old rats were placed into organ culture and incubated in the absence or presence of AMH, either alone or in combination with known stimulators of follicle transition, including basic fibroblast growth factor (bFGF), kit ligand (KITL), or keratinocyte growth factor (KGF). Following 10 days of culture, the ovaries were sectioned, stained, and morphologically evaluated to determine the percentage of primordial versus developing follicles. As previously demonstrated, AMH treatment decreased primordial to primary follicle transition. Interestingly, AMH inhibited the stimulatory actions of KITL, bFGF, and KGF. Therefore, AMH can inhibit the basal and stimulated development of primordial follicles. To investigate the mechanism of AMH actions, the influence AMH has on the ovarian transcriptome was analyzed. AMH treatment when compared with controls was found to alter the expression of 707 genes. The overall effect of AMH exposure is to decrease the expression of stimulatory factors, increase the expression of inhibitory factors, and regulate cellular pathways (e.g. transforming growth factor beta signaling pathway) that result in the inhibition of primordial follicle development. Analysis of the regulatory factors and cellular pathways altered by AMH provides a better understanding of the molecular control of primordial follicle development. Experiment Overall Design: RNA samples from two control groups (pooled untreated cultured ovaries) are compared to two treated groups (pooled cultured ovaries treated with AMH)

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Functional germ line stem cells do not exist in adult mammalian ovaries

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