Dataset Information

MicroRNA-34c from follicular fluid-derived extracellular vesicles is a modulator of embryo quality

ABSTRACT: Oocyte competence is progressively acquired during follicle growth, with large follicles containing more often competent oocytes than small follicles. A competent oocyte is able to mature in vitro, be fertilized, and develop to the blastocyst stage. Extracellular vesicles (EVs) and their cargo micro RNAs (miRNAs) are critical in mediating intercellular communication inside the follicle and facilitating oocyte maturation. Whether follicular EVs are also involved in the acquisition of oocyte competence in the growing follicle remains unclear. Here, we have been using an individual culture system to distinguish follicular fluid and their corresponding bovine oocytes as competent or noncompetent. We used a qEV single-column method to isolate EVs from follicular fluid. RNA sequencing revealed 16 differentially expressed (DE) miRNAs in EVs from follicular fluid derived from either competent or noncompetent oocytes . Among the up-regulated miRNAs, we selected bta-miR-34c to further validate its effect on oocyte competence during in vitro maturation using mimics and inhibitors. By supplementing miR-34c mimics to the oocyte maturation medium, we could significantly improve blastocyst quality, as evidenced by higher cell numbers, while supplementation of miR-34c inhibitors produced the opposite effect. Taken together, the up-regulation of miR-34c in EVs derived from follicular fluid from competent oocytes is indicative of its regulatory role, and, when added during in vitro maturation of unselected oocytes, is able to increase total cell number and inner cell mass of resulting embryos, thus contributing to the development of healthy offspring

ORGANISM(S): Bos taurus

PROVIDER: GSE250281 | GEO | 2024/09/03

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Oocyte competence for early embryo development relies on intercellular communication between the maturing oocyte and preovulatory follicle. Preovulatory follicle maturity, as indicated by serum estradiol concentration or follicle diameter, has previously been linked to pregnancy, follicular fluid metabolites, cumulus-oocyte metabolism, and oocyte competency for embryo development. Such relationships indicate metabolic and developmental programming of the oocyte based on the preovulatory follicle’s physiological status, but downstream impacts on the molecular signature of blastocysts have not been examined. We hypothesized that supplementing maturing oocytes with follicular fluid originating from preovulatory follicles of greater or lesser maturity would impact the transcriptome of resulting blastocysts and indicate metabolic programming of the embryo that originated from the oocyte’s maturation environment. The objective was to investigate the effect of follicle maturity on metabolic preparation of the oocyte by examining the functional genome of blastocysts originating from oocytes matured in the presence of follicular fluid from preovulatory follicles of greater or lesser maturity. In vitro maturing oocytes were supplemented with follicular fluid collected from preovulatory follicles of greater or lesser maturity. Following identical embryo culture procedures, RNA-sequencing was performed on pools of 2 blastocysts (Greater, n = 12; Lesser, n = 15; all with stage code = 7 and quality code = 1). A total of 12,310 gene transcripts were identified in blastocysts after filtering to remove lowly abundant transcripts. There were 113 transcripts that differed in abundance between blastocysts originating from oocytes matured in greater versus lesser maturity follicular fluid (eFDR < 0.01). Although no pathways were significantly enriched with differentially abundant transcripts, transcriptome profiles suggested improved Wnt/β-catenin signaling, metabolism, and protection from oxidative stress in blastocysts derived from oocytes matured in greater maturity follicular fluid, while unregulated cell growth presented in blastocysts resulting from the lesser follicle maturity treatment.

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

Dataset Information

MicroRNA-34c from follicular fluid-derived extracellular vesicles is a modulator of embryo quality

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets