Androgen Receptor mRNA Expression in the Rhesus Monkey Ovary

Abstract

Immunocytochemical detection of androgen receptors (ARs) in several compartments of the macaque ovary, including the germinal epithelium, follicle, and corpus luteum, suggests a role for androgens in modulating ovarian function via the classical receptor-mediated pathway. To examine AR mRNA expression in the rhesus monkey ovary, total RNA was isolated from whole ovaries, the germinal epithelium-enriched cortical and medullary compartments of the ovary, and corpora lutea from early (d 3-5), mid (d 6-8), mid-late (d 10-12), and late (d 13-15) stages of the luteal phase of the menstrual cycle. RNA was also obtained from luteinized granulosa cells from monkeys receiving gonadotropin treatment to stimulate the development of multiple ovarian follicles. After reverse transcription of total RNA using oligo-dT as a primer, polymerase chain reaction (PCR) was used to amplify a unique 329 bp segment of the monkey AR hormone-binding region. Reverse transcriptase (RT)-PCR products of the expected size were detected in all ovarian and control tissues. Sequence analysis of the AR cDNA from the macaque ovary revealed 99% nucleotide homology and 100% predicted amino acid homology to the cDNA for the hormone-binding region of human AR. Northern analysis demonstrated the presence of a major AR mRNA species at 9.5 kb in corpus luteum, luteinized granulosa cells, and prostate, with additional bands detected in the corpus luteum and prostate at 7.9 and 3.4 kb, respectively. A sensitive RNase protection assay was used to examine AR mRNA levels in ovarian tissues and showed AR mRNA expression throughout the life-span of the corpus luteum. Thus, detection of AR mRNA in the primate ovary, including the periovulatory follicle and corpus luteum, supports the concept that these tissues are targets for receptor-mediated androgen action during the menstrual cycle.

Role of androgens in normal and pathological ovarian function

“…AR mRNA is present in primate corpora lutea (CL) (Duffy et al 1999). During the early luteal phase of a cycle, developing and even regressing primate CL exhibit AR immunostaining (Hild-Petito et al 1991).…”

Androgen actions in the ovary: balance is key

“…Studies across species reported that ARs are expressed in theca cells, granulosa cells (GCs), and the oocyte of the follicle and throughout most stages of follicular development (Horie et al 1992, Chadha et al 1994, Hirai et al 1994, Suzuki et al 1994, Tetsuka & Hillier 1996, Szoltys & Slomczynska 2000, Slomczynska & Tabarowski 2001, Gill et al 2004, Hampton et al 2004, Juengel et al 2006. However, their expression patterns may differ between ovarian cell types (Tetsuka & Hillier 1996, Szoltys & Slomczynska 2000 and in most species, AR is abundant in the preantral/antral stages of follicular development but declines as follicles mature to the preovulatory stage (Horie et al 1992, Chadha et al 1994, Suzuki et al 1994, Duffy et al 1999, Hampton et al 2004, Juengel et al 2006. In fact, AR is expressed in cell-specific regions of human ovarian follicles at all stages of follicular development beyond the primordial phase (Rice et al 2007).…”

Vascular Endothelial Growth Factor (VEGF) Production by the Monkey Corpus Luteum During the Menstrual Cycle: Isoform-Selective Messenger RNA Expression In Vivo and Hypoxia-Regulated Protein Secretion In Vitro1

“…Serum estradiol and P 4 levels were determined using ELISA (Elecsys 2010; Roche Diagnostics, Indianapolis, IN) as described previously [18]. The first day of low (Ͻ100 pg/ml) serum estradiol levels following the midcycle estradiol peak corresponds with the day after the LH surge and, therefore, was considered to be Day 1 of the luteal phase [19].…”