Oocyte maturation is a critical stage in embryo development and female reproduction, and is regulated by adipokines- hormones produced by adipose tissue. Omentin-1 (ITLN1) is an adipokine known to regulate ovarian follicle function, however, its role in oocyte maturation remains unclear. This study investigated ITLN1 expression in porcine oocytes and cumulus cells before and after in vitro maturation (RT-qPCR, Western blotting, immunofluorescence) and the effect of ITLN1 on: oocyte maturation by assessing the percentage of oocytes in metaphase II (M2) (DAPI) and progesterone (P4) secretion (ELISA); glucose metabolism, including glucose transporter type 4 (GLUT4) expression, glucose and lactate secretion, and mitochondrial function (Seahorse Test); lipid metabolism, as assessed by lipid droplet area (Nile Red) and adipose differentiation-related protein and adipose triglyceride lipase (ATGL) expression; extracellular signal-regulated kinase 1/2 (ERK1/2) and AMP-activated protein kinase alpha 1 (PRKAA1) phosphorylation, which mediating the effects of ITLN1 on oocyte maturation. Moreover, the effect of ITLN1 on preimplantation embryo development was also examined. ITLN1 expression increased after maturation in both oocytes and cumulus cells, with strong immunofluorescence signals in both cells. ITLN1 increased oocyte maturation to the M2 stage and P4 secretion. It enhanced glucose metabolism by upregulating GLUT4 and reducing glucose and lactate levels, improved mitochondrial function, reduced lipid droplet area, upregulated ATGL expression, and stimulated ERK1/2 and PRKAA1 phosphorylation. However, ITLN1 had no significant effect on preimplantation embryo development. In conclusion, ITLN1 positively regulates porcine oocyte maturation in vitro through coordinated modulation of energy metabolism, mitochondrial activity, lipid utilization, and ERK1/2-PRKAA1 signalling.
K. Kubicka, N. Respekta-Długosz, Patrycja Kurowska et al.· Reproduction· 0 citations
Adropin is a new protein that regulates energy homeostasis. The serum and follicular fluid (FF) levels of adropin are decreased in women with polycystic ovarian syndrome (PCOS); however, its role in ovarian function is unknown. The aims were to determine the expression of adropin and its receptor G protein-coupled receptor 19 (GPR19) in human granulosa cells (GC), its immunolocalization and its in vitro effects on GC function. Blood plasma, FF and GC samples were obtained from normal weight, obese and diagnosed with or without PCOS women (n=8). The in vitro effects of adropin on GC proliferation, apoptosis, cell cycle progression, steroidogenesis were analyzed. The results revealed that adropin plasma concentration was decreased in obese patients, with a similar reduction observed in obese patients with PCOS, while GPR19 expression was decreased in the GC of obese and PCOS women, as well as in obese PCOS patients. We noted that in all investigated patients groups adropin reduced GC proliferation and cell cycle progression, negatively influenced steroidogenesis enzyme levels and promoted apoptosis. Such disruptions in GC function are likely to impair ovarian follicular maturation and contribute to the subfertility commonly observed in PCOS. Such alterations may ultimately affect oocyte competence and ovarian responsiveness, parameters that are clinically relevant for in vitro fertilization outcomes. Our findings suggest that adropin may act as a novel regulator of ovarian function and could contribute to the pathophysiology of PCOS, highlighting its potential clinical value as a marker of altered ovarian follicular function in affected women.
Patrycja Kurowska, M. Dawid, N. Respekta-Długosz et al.· Journal of Endocrinology· 0 citations
Asprosin is a profibrillin-1-derived protein hormone involved in energy metabolism, glucose homeostasis, and appetite. Although asprosin can cross the blood-brain barrier and act within the central nervous system, its direct role in hippocampal neurons remains unclear. This study examined asprosin system components and the effects of asprosin on metabolic activity, proliferation, reactive oxygen species (ROS) production, cell cycle progression, apoptosis, and intracellular signaling in mouse hippocampal HT-22 cells. Asprosin, Furin, and Olfr734 expression was assessed by Western blot and immunocytochemistry, whereas intracellular and secreted asprosin levels were quantified by ELISA. Cells were treated with asprosin over 0.01-100 nM, and low nanomolar doses were selected for detailed cell cycle, apoptosis, and signaling analyses. Metabolic activity, proliferation, ROS production, cell cycle distribution, and apoptosis were evaluated using alamarBlue, bromodeoxyuridine incorporation, ROS detection, and flow cytometry assays. Selected ROS-, cell cycle-, and apoptosis-related markers were analyzed by reverse transcription quantitative polymerase chain reaction and Western blot. PKA and ERK1/2 involvement was assessed by phosphorylation analysis and pharmacological inhibition. HT-22 cells expressed Asprosin, Furin, and Olfr734. Asprosin regulated cellular responses in a dose- and time-dependent manner: low nanomolar doses generally supported metabolic and proliferative activity, whereas 10 and 100 nM reduced these parameters. Asprosin did not markedly increase total ROS production but modulated Hmox1 and Sod2 expression. It also altered cell cycle distribution, apoptosis-related parameters, PKA and ERK1/2 phosphorylation, and inhibitor-sensitive expression of Ccne1, Bax, and Bcl2. These findings identify, for the first time, a functional asprosin system in mouse HT-22 hippocampal neurons.
N. Respekta-Długosz, Renata Głowaczewska, Dominika Wachowska et al.· Neuroscience· 0 citations