Norepinephrine, Active Norepinephrine Transporter, and Norepinephrine-Metabolism Are Involved in the Generation of Reactive Oxygen Species in Human Ovarian Granulosa Cells

Saller, S.; Merz-Lange, Julia; Raffael, Sandra; Hecht, S.; Pavlik, R.; Thaler, Christian J.; Berg, Dieter; Berg, U.; Kunz, Lars; Mayerhofer, Artur

doi:10.1210/en.2011-1769

Cited by 49 publications

(73 citation statements)

References 45 publications

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“…As recently found in human FF [3], macaque FF NE levels exceeded those in serum, which were below the detection limit of 0.2 ng/ml in monkeys.…”

Section: Resultsmentioning

confidence: 49%

“…Thus, in FF of large monkey follicles, NE accumulated and reached levels that are much higher than serum levels. This is comparable to the situation in the human ovary [3]. The reason and mechanisms for accumulation in the FF remain unknown, but may be related to the expression of NE- and DA-transporters and storage-capacities of GCs with respect to catecholamines [3,4,41] and to the ability of oocytes to metabolize DA to NE [42].…”

Section: Discussionmentioning

confidence: 66%

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

Merz

Saller

Kunz³

et al. 2015

J Ovarian Res

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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.

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“…As recently found in human FF [3], macaque FF NE levels exceeded those in serum, which were below the detection limit of 0.2 ng/ml in monkeys.…”

Section: Resultsmentioning

confidence: 49%

Section: Discussionmentioning

confidence: 66%

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

Merz

Saller

Kunz³

et al. 2015

J Ovarian Res

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“…FF, was frozen at -20°C until use for Western blot and proNGF ELISA studies. Washed cells were re-suspended in culture media supplemented with penicillin (100 U/ml), streptomycin (100 μg/ml) and 10% (v/v) fetal calf serum (FCS) (all from PAA) as previously described [26-30]. Experiments were performed on day 2 or 3 of cell culture, unless otherwise indicated.…”

Section: Methodsmentioning

confidence: 99%

“…Collection of tissues was approved by the Oregon National Primate Research Center Institutional Animal Care and Use Committee. Sections (4 μm) were cut from all paraffin-embedded samples and were used for immunohistochemistry, as described [30]. In brief, sections were deparaffinized, subjected to antigen retrieval by heating in citrate buffer (1.8 mM citric acid, 8.2 mM sodium citrate, pH 6.0) at 90°C for 30 min and endogenous peroxidase activity was blocked.…”

Section: Methodsmentioning

confidence: 99%

Pro-nerve growth factor in the ovary and human granulosa cells

Meinel

Blohberger

Berg

et al. 2015

Hormone Molecular Biology and Clinical Investigation

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Background Pro-nerve growth factor must be cleaved to generate mature NGF, which was suggested to be a factor involved in ovarian physiology and pathology. Extracellular proNGF can induce cell death in many tissues. Whether extracellular proNGF exists in the ovary and may play a role in the death of follicular cells or atresia was unknown. Material and Methods Immunohistochemistry of human and Rhesus monkey ovarian sections was performed. IVF-derived follicular fluid and human granulosa cells were studied by RT-PCR, qPCR, Western blotting, ATP- and caspase-assays. Results and Conclusions Immunohistochemistry of ovarian sections identified proNGF in granulosa cells and Western blotting of human isolated granulosa cells confirmed the presence of proNGF. Ovarian granulosa cells thus produce proNGF. Recombinant human proNGF even at high concentrations did not affect the levels of ATP or the activity of caspase 3/7, indicating that in granulosa cells proNGF does not induce death. In contrast, mature NGF, which was detected previously in follicular fluid, may be a trophic molecule for granulosa cells with unexpected functions. We found that in contrast to proNGF, NGF increased the levels of the transcription factor early growth response 1 and of the enzyme choline acetyl-transferase. A mechanism for the generation of mature NGF from proNGF in the follicular fluid may be extracellular enzymatic cleavage. The enzyme MMP7 is known to cleave proNGF and was identified in follicular fluid and as a product of granulosa cells. Thus the generation of NGF in the ovarian follicle may depend on MMP7.

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“…Due to that oxidative 37 stress may be classified as basal oxidative stress (BOS), low intensity oxidative stress (LOS), intermediate 38 intensity oxidative stress (IOS), and high intensity oxidative stress (HOS). Another classification of poten- 39 tial interest may differentiate three categories such as mild oxidative stress (MOS), temperate oxidative 40 stress (TOS), and finally severe (strong) oxidative stress (SOS). Perspective directions of investigations in 41 the field include development of sophisticated classification of oxidative stresses, accurate identification 42 of cellular ROS targets and their arranged responses to ROS influence, real in situ functions and operation 43 of so-called ''antioxidants'', intracellular spatiotemporal distribution and effects of ROS, deciphering of 44 molecular mechanisms responsible for cellular response to ROS attacks, and ROS involvement in realiza- 45 tion of normal cellular functions in cellular homeostasis.…”

mentioning

confidence: 99%