Expression of the beta-2 adrenergic receptor (ADRB-2) in human and monkey ovarian follicles: a marker of growing follicles?

Abstract

BackgroundADRB-2 was implicated in rodent ovarian functions, including initial follicular growth. In contrast, ADRB-2 expression and function in nonhuman primate and human ovary were not fully known but innervation and significant levels of norepinephrine (NE), which is a ligand at the ADRB-2, were reported in the ovary.MethodsWe studied expression of ADRB-2 in human and rhesus monkey ovary (RT-PCR, immunohistochemistry; laser micro dissection) and measured levels of norepinephrine (NE; ELISA) in monkey follicular fluid (FF). 3D cultures of monkey follicles (4 animals) were exposed to NE or the ADRB-2 agonist isoproterenol (ISO), and follicular development (size) was monitored. Upon termination expression of ADRB-2, FSH receptor and aromatase genes were examined.ResultsImmunohistochemistry and RT-PCR of either human follicular granulosa cells (GCs) obtained by laser micro dissection or isolated monkey follicles revealed ADRB-2 in GCs of primordial, primary, secondary and tertiary follicles. Staining of GCs in primordial and primary follicles was intense. In large preantral and antral follicles the staining was heterogeneous, with positive and negative GCs present but GCs lining the antrum of large follicles were generally strongly immunopositive. Theca, interstitial, and ovarian surface epithelial cells were also positive. NE was detected in FF of preovulatory antral monkey follicles (0.37 + 0.05 ng/ml; n = 7; ELISA) but not in serum. We examined preantral follicles ranging from 152 to 366 μm in diameter in a 3D culture in media supplemented with follicle stimulating hormone (FSH). Under these conditions, neither NE, nor ISO, influenced growth rate in a period lasting up to one month. Upon termination of the cultures, all surviving follicles expressed aromatase and FSH receptors, but only about half of them also co-expressed ADRB-2. The ADRB-2 expression was not correlated with the treatment but was positively correlated with the follicular size at the beginning and at the end of the culture period. Hence, expression of ADRB-2 was found in the largest and fastest-in vitro growing follicles.ConclusionsThe results imply ADRB-2-mediated actions in the development of primate follicles. Drugs interfering with ADRB-2 are used to treat medical conditions and may have unexplored effects in the human ovary.

Antioxidant Intervention Attenuates Aging-Related Changes in the Murine Ovary and Oocyte

“…Following antioxidant administration, the murine ovarian transcriptome reflected an environment with enhanced cell survival and β-adrenergic signaling, decreased apoptotic signaling proinflammatory pathways, and altered antioxidant signaling compared to control aged mice. Upregulated β-adrenergic signaling may contribute to improved ovarian function through changes in follicular development and hormone secretion [37][38][39], particularly since ovarian sympathetic nerve activity may decline with ovarian aging [40]. The anti-inflammatory effects of antioxidant treatment are consistent with previous reports [41].…”

Intrafollicular and Systemic Dopamine, Noradrenaline and Adrenaline Concentrations in Cycling Mares

“…Metanephrine, a metabolite of DA and NA in FF and granulosa cells, also provides clear evidence of the follicular metabolism of catecholamines [ 25 ]. In addition, a ligand (NA in FF) and its receptor (ADRB-2 in granulosa cells) co-exist in the large antral follicles of primates [ 39 ]. Likewise, NA can be synthesized by the neuron-like cells in the ovary and oocytes.…”

Gonadotropin Ratio Affects the In Vitro Growth of Rhesus Ovarian Preantral Follicles

“…To the best of our knowledge, our study is the first to show that high dose gonadotropin treatment shortens the duration of IVFG using rhesus preantral follicles (220–380 µm) and that the gonadotropin ratio affects the IVFG efficiency in rhesus models as well as in murine models. As aforementioned, previous reports employed low dose, long-term culture for the IVFG of secondary follicles sized less than 366 µm in diameter, 8 9 or high dose, short-term culture for the IVFG of small antral follicles sized larger than 500 µm in diameter. 10 11 …”

“…Gonadotropin treatment is also essential in the IVFG of animal models. In many studies, only FSH was used, 8–9 11 and in others a combination of FSH and LH was used. 10 Our data showed that the FSH:LH of the 2:1 group was superior to that of the 1:1 group ( figure 3 ).…”

“…Most of the previous reports used rodents at the adolescent or pre-reproductive period, for instance 12–14-day-old mice, for IVFG of preantral ovarian follicles. 12 14 15 In previous studies using rhesus monkeys, they retrieved ovaries from the animals of 7–12 years for small antral follicle culture 10 11 and from those of 4–17 years for secondary follicle culture; 8 9 both ages correspond to mid to late reproductive period since their adolescence begins at around 2 years. Our animals were younger than those in previous reports, that is, 4–5 years and two 8–9 years.…”