Abstract

Induced ovulation of small pre-ovulatory follicles reduced pregnancy rates, embryo survival, day seven embryo quality, and successful embryo cleavage in beef cows undergoing fixed-time artificial insemination. RNA-sequencing of oocytes and associated cumulus cells collected from pre-ovulatory follicles 23 h after gonadotropin-releasing hormone (GnRH) administration to induce the pre-ovulatory gonadotropin surge suggested reduced capacity for glucose metabolism in cumulus cells of follicles ≤11.7 mm. We hypothesized that the follicular fluid metabolome influences metabolic capacity of the cumulus-oocyte complex and contributes to reduced embryo cleavage and quality grade observed following induced ovulation of small follicles. Therefore, we performed a study to determine the correlation between pre-ovulatory follicle diameter and follicular fluid metabolome profiles in lactating beef cows (Angus, n = 130). We synchronized the development of a pre-ovulatory follicle and collected the follicular contents approximately 20 h after GnRH administration. We then performed ultra-high performance liquid chromatography—high resolution mass spectrometry (UHPLC-HRMS) metabolomic studies on 43 follicular fluid samples and identified 38 metabolites within pre-ovulatory follicles of increasing size. We detected 18 metabolites with a significant, positive correlation to follicle diameter. Individual and pathway enrichment analysis of significantly correlated metabolites suggest that altered glucose and amino acid metabolism likely contribute to reduced developmental competence of oocytes when small pre-ovulatory follicles undergo induced ovulation.

Highlights

  • Decreased pre-ovulatory follicle diameter at the time of pharmacological induction of ovulation was associated with lower pregnancy rates and/or reduced embryo survival in beef cattle undergoing fixed-time artificial insemination (FTAI) [1,2,3,4]

  • Though studies of the follicular fluid metabolome have been performed in humans and dairy cows, little focus has been placed on the impact of follicle diameter at induced ovulation on the follicular fluid and resulting cellular functions of the granulosa cells or cumulus-oocyte complex in beef cows

  • We hypothesized that size-dependent changes in the follicular fluid’s metabolome influence metabolic capacity of the cumulus-oocyte complex that contribute to reduced embryo cleavage and quality grade observed following induced ovulation of small pre-ovulatory follicles

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Summary

Introduction

Decreased pre-ovulatory follicle diameter at the time of pharmacological induction of ovulation was associated with lower pregnancy rates and/or reduced embryo survival in beef cattle undergoing fixed-time artificial insemination (FTAI) [1,2,3,4] Such reductions in fertility were likely the result of poor preparation of the maternal environment for pregnancy establishment and the ovulation of an oocyte with reduced developmental competency [2,5,6,7]. Studies that investigate the relationship between pre-ovulatory follicle size and the follicular fluid milieu are essential to further understanding reduced oocyte developmental competence and subsequent fertility when beef cows are induced to ovulate a small pre-ovulatory follicle. We hypothesized that size-dependent changes in the follicular fluid’s metabolome influence metabolic capacity of the cumulus-oocyte complex that contribute to reduced embryo cleavage and quality grade observed following induced ovulation of small pre-ovulatory follicles. We utilized ultra-high performance liquid chromatography—high resolution mass spectrometry (UHPLC-HRMS) to identify metabolites within pre-ovulatory follicles of increasing size and determined metabolites with a significant correlation to follicle diameter

Animal Data
Follicular Fluid Processing
UHPLC-HRMS Metabolomics
Primary Data Analysis
Statistical Analyses
Conclusions
Full Text
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