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Dynamic modeling of the murine oocyte and somatic cell metabolome during follicle development

ABSTRACT: Explore in vivo ovarian follicle transcriptomics from primordial follicles to the antral stage. Ovarian follicles were isolated from CD-1 mice. Entire ovaries were collected at post-natal day 3 and day 4 to collect primordial follicles. Primary follicles (70-90 µm in diameter), two-layered secondary follicles (100-130 µm), multi-layer secondary follicles (150‒180 µm), and pre-antral follicles (200‒300 µm) were mechanically isolated from post-natal day 10, 12, 16, and 18 ovaries, respectively. Antral follicles (400‒600 µm) were mechanically isolated from pregnant mare serum gonadotropin (PMSG)-treated mice ovaries at post-natal day 20. Follicles were then aspirated and combined per ovarian follicle maturation stage (e.g., primary, two-layered secondary). Three different samples were collected from each pooled follicular stage for transcriptomic analysis. RNA was purified and hybridized in MouseRef-8 v2.0 Expression BeadChip Kit (Illumina, San Diego, CA), as previously described (Skory, Bernabe et al., 2013).

ORGANISM(S): Mus musculus

PROVIDER: GSE97902 | GEO | 2017/04/18

SECONDARY ACCESSION(S): PRJNA383162

REPOSITORIES: GEO

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Project description:Dibutyl phthalate (DBP), di-2-ethylhexyl phthalate (DEHP), and benzyl butyl phthalate (BBP) are three phthalates commonly found in consumer products, including the plastic coating of pharmaceuticals and personal care products. Folliculogenesis, a tightly regulated process occurring in the ovary, is the maturation of an immature primordial follicle to a mature antral follicle. Follicles house the oocyte and antral follicles specifically play a crucial role in ovarian steroidogenesis and ovulation. DBP, BBP, and DEHP have been associated with inhibited antral follicle growth in vitro, decreased ovulation rates in vitro, and decreased antral follicle counts in women. However, little is known about the effects of a three-phthalate mixture on antral follicles in vivo. The objective of this study was to evaluate the effects of a human relevant mixture of DBP, BBP, and DEHP on ovarian follicles through proteome profiling analysis. CD-1 female mice (60 days old) were pipet fed tocopherol stripped corn oil (vehicle control) only or a phthalate mixture (52% DBP, 36% DEHP, and 12% BBP dissolved in vehicle) which modeled human follicular fluid concentrations. The mice were treated with 32µg/kg/day (PHT Mix 32; cumulative estimate in general population) and 500µg/kg/day (PHT Mix 500; cumulative estimate in occupationally exposed individuals) for 10 consecutive days. Proteome profiling of antral follicles (>250µm) was performed via label-free tandem mass spectrometry. A total of 5,417 antral follicle proteins were identified in the three groups, of which 194 were differentially abundant between the vehicle and PHT Mix 32 group, and 136 between the vehicle and PHT Mix 500 group. Gene ontology analysis revealed that the two treatments of the phthalate mixture upregulate and downregulate distinctive processes, supporting non-monotonic effects of phthalates on the antral follicle proteome. Taken together, these results reveal that a human relevant mixture of DBP, BBP, and DEHP alters the antral follicle proteome and merits further evaluation to elucidate the molecular mechanisms by which phthalates cause negative reproductive outcomes.

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Dynamic modeling of the murine oocyte and somatic cell metabolome during follicle development

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