Glutamine and norepinephrine in follicular fluid synergistically enhance the antioxidant capacity of human granulosa cells and the outcome of IVF-ET

Abstract

An increasing number of studies demonstrate that changes in neurotransmitters metabolic levels in follicular fluid are directly related to oocyte maturation, fertilization, the quality of embryo and pregnancy rates. However, the relationship between the intra-follicular neurotransmitters and the function of granulosa cells (GCs), and the outcome of in vitro fertilization-embryo transfer (IVF-ET) is not clear. Human follicular fluid and cumulus GCs were harvested from large follicles obtained from patients undergoing IVF. Neurotransmitters and steroid hormones in follicular fluid were measured through liquid chromatography-tandem mass spectrometry (LC–MS/MS) and high-performance liquid chromatography-mass spectrometry (HPLC–MS/MS). Based on the content of glutamine (Gln) in follicular fluid, the samples were divided into two groups: high Gln level group and low Gln level group. The expression of proliferation-, steroidogenesis- and antioxidant-related genes in GCs was detected by qRT-PCR. In vitro, KGN cells were used to further verify the effects of Gln and NE on GCs function. Primary and secondary outcomes were the number of mature and retrieved oocytes, and the ratio of high-quality embryos, respectively. Gln (46.75 ± 7.74 μg/mL) and norepinephrine (NE, 0.20 ± 0.07 μg/mL) were abundant neurotransmitters in follicular fluid, and exhibited a significantly positive correlation (R = 0.5869, P < 0.005). In high Gln level group, the expression of proliferation, steroidogenesis and antioxidant-related genes in GCs were higher than those in low Gln level group, and the contents of estriol and E2 in follicular fluid were more abundant. Moreover, the concentrations of Gln and NE in follicular fluid showed significantly positive correlation with IDH1 expression in GCs (R = 0.3822, R = 0.4009, P < 0.05). Importantly, a significantly positive correlation was observed between IDH1 expression in GCs and the ratio of higher-quality/cleaved embryos (R = 0.4480, P < 0.05). In vitro studies further demonstrated that Gln and NE played synergistically function in improving GCs proliferation and E2 production by upregulating IDH1 expression. These data demonstrate that Gln and NE in follicular fluid might play significant positive roles in GCs function, and may be potential predictors for selecting optimal quality oocytes and evaluating the quality of embryonic development.

Differential effects of follicle-stimulating hormone glycoforms on the transcriptome profile of cultured rat granulosa cells as disclosed by RNA-seq

“…CREB) exhibited underexpression at this time point, regardless of their cellular or pathway position. The exception was AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) receptor, which is stimulated by glutamic acid present in follicular fluid [52] and in turn regulates signaling molecules (presumably via calcium influx) involved in ovulation [53]; AMPAR showed overexpression with all treatments. At this time, signaling was also very similar between FSH 18/21 and eq FSH with the exception of GPCRs for glycoprotein hormones and c-Jun N-terminal kinase (JNK; a kinase involved in cell cycle progression, mitosis and apoptosis [54, 55]) signaling overexpression stimulated by eq FSH.…”

“…CREB) exhibited underexpression at this time point, regardless of their cellular or pathway position. The exception was AMPA (αamino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) receptor, which is stimulated by glutamic acid present in follicular fluid [52] and in turn regulates signaling molecules (presumably via calcium influx) involved in ovulation [53]; AMPAR showed overexpression with all treatments.…”

“…1). In glycoprotein hormones, the oligosaccharide in position Asn 52 is involved in the activation of the receptor/signal transducer (G protein) system and biological response [3][4][5][6], an effect mediated through the stabilization of the conformation of the FSH dimer bound to the FSH receptor (FSHR) [7][8][9]. Meanwhile, glycans attached to FSH play a major role in defining the circulatory half-life and in vivo bioactivity of the gonadotropin [10],…”

Norepinephrine induces anoikis resistance in high-grade serous ovarian cancer precursor cells

“…In light of new evidence that peripheral nerves can initiate and potentiate tumor growth through delivery of neurotransmitters, such as NE ( 36 , 41 , 65 , 66 ), we considered whether NE may play a role in the progression of HGSC from the FT. We utilized immortalized human FTE cell lines with reduced TP53 expression (shTP53) ( 67 , 68 ), characteristic of HGSC precursor lesions ( 13 ), and exposed them to a physiologically relevant dose of NE ( 69 ) that is also commonly used in relevant experimental reports ( 54 , 70 ). Overall, we demonstrate that NE enhances FTE cell spheroid formation and subsequent viability in ULA culture conditions, enabling adhesion of these aggregates to the OSE.…”

A Pilot Study on Biochemical Profile of Follicular Fluid in Breast Cancer Patients

“…A recent study by Wang et al, has shown that Gln, synergistically with norepinephrine, promotes estrogen synthesis in FF of IVF patients, and increases the expression of antioxidant genes, as antioxidant gene IDH1, glutathione peroxidase, and Nrf2. Besides, high levels of Gln improve the function of granulose cells in vitro [ 55 ]. On the other hand, it cannot be excluded that the decrease of Gln found in patients with BC is the result of a compensatory mechanism to attenuate oxidative stress [ 56 ].…”