Chronically elevated androgen and/or consumption of a Western-style diet impairs oocyte quality and granulosa cell function in the nonhuman primate periovulatory follicle
To investigate the impact of chronically elevated androgens in the presence and absence of an obesogenic diet on oocyte quality in the naturally selected primate periovulatory follicle.
Rhesus macaques were treated using a 2-by-2 factorial design (n = 10/treatment) near the onset of menarche with implants containing either cholesterol (C) or testosterone (T, 4–5-fold increase above C) and a standard or “Western-style” diet alone (WSD) or in combination (T+WSD). Following ~ 3.5 years of treatment, females underwent controlled ovulation (COv, n = 7–10/treatment) cycles, and contents of the naturally selected periovulatory follicle were aspirated. Follicular fluid (FF) was analyzed for cytokines, chemokines, growth factors, and steroids. RNA was extracted from luteinizing granulosa cells (LGCs) and assessed by RNA-seq.
Only healthy, metaphase (M) I/II-stage oocytes (100%) were retrieved in the C group, whereas several degenerated oocytes were recovered in other groups (33–43% of T, WSD, and T+WSD samples). Levels of two chemokines and one growth factor were reduced (p < 0.04) in FF of follicles with a MI/MII oocyte in WSD+T (CCL11) or T and WSD+T groups (CCL2 and FGF2) compared to C and/or WSD. Intrafollicular cortisol was elevated in T compared to C follicles (p < 0.02). Changes in the expression pattern of 640+ gene products were detected in LGC samples from follicles with degenerated versus MI/MII-stage oocytes. Pathway analysis on RNAs altered by T and/or WSD found enrichment of genes mapping to steroidogenic and immune cell pathways.
Female primates experiencing hyperandrogenemia and/or consuming a WSD exhibit an altered intrafollicular microenvironment and reduced oocyte quality/competency, despite displaying menstrual cyclicity.
KeywordsAndrogen Follicular cytokines Nonhuman primate Oocyte quality Periovulatory follicle Western-style diet
The outstanding efforts of the NCTRI NHP Core members Emily Mishler, M.S., Corrine Wilcox, B.S., Kise Bond, P.S.M., and Andrea Calhoun, M.S. under the direction of Ov Slayden, Ph.D. as well as Diana Takahashi, M.S., were critical for the successful execution of all protocols and are greatly appreciated. In addition, the efforts of the staff of the ONPRC Endocrine Technology Support Core contributed to the success of this project. RNA sequencing was performed by the OHSU Massively Parallel Sequencing Shared Resource. Analysis of the RNA-seq data was performed by the ONPRC’s Biostatistics & Bioinformatics Core under the direction of Lucia Carbone, Ph.D. and Suzanne Fei, Ph.D. with statistical analyses performed by Lina Gao, Ph.D. and Byung Park, Ph.D. We greatly appreciate the core’s analysis pipelines that utilize OHSU’s Exacloud compute cluster.
Research reported in this publication was supported by the Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD) of the National Institutes of Health (NIH) under Award Number P50HD071836 (to RLS). Additional funding was provided by NIH Award Number P51OD011092 (Support for National Primate Research Center and Cores).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
All procedures involving rhesus monkeys were reviewed and approved by the ONPRC/Oregon Health and Science University (OHSU) Institutional Animal Care and Use Committee (IACUC) in accordance with the U.S. Public Health Service (PHS) Policy on Humane Care and Use of Laboratory Animals.
The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
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