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• W2738864970 abstract "Prorenin levels are increased in follicular fluid after LH surge. Recently, our group identified the presence of (pro)renin receptor [(P)RR] in bovine cumulus-oocyte complex (COC). Our aim was to evaluate the effect of prorenin in resumption of meiosis and in the levels of cAMP and cGMP in cumulus cells (CCs) and oocytes. For this, bovine ovaries were obtained in a local abattoir and approximately 20 COCs/ treatment were cultured in 200μl of TCM-199 for 15 hours at 39°C. The treatments were: positive control, negative control (FSK; 200μM), FSK and prorenin (10-10M), and FSK, prorenin plus aliskiren (ALK; direct renin inhibitor, 10-7M). Nuclear maturation was considered when oocytes reached the stage of metaphase I (MI) using10mg/mL of bisbezimide (Hoechst 33342) in fluorescence microscope. Statistical analysis was performed using SAS with significance of 5%. The rate of oocytes that reached MI was greater in the group treated with FSK plus prorenin (38.39%) compared with negative control (18.92%), and FSK, prorenin and ALK groups (8.68%, P<0.05). To determine the effect of prorenin on cAMP and cGMP concentrations in oocytes and CCs, COCs were cultured for 6 and 15 hours under the same conditions previously described and distributed as follows: positive control, negative control (FSK), prorenin and FSK plus prorenin. The concentrations of cAMP and cGMP were measured on CC (n=60 COCs) and oocytes (n=50) after 6h of culture using cAMP EIA kit (No. 581 001; Cayman Chemical) and cGMP EIA kit (No. 581 021; Cayman Chemical) according to manufacturer’s instructions. Data were tested for normal distribution using the Shapiro-Wilk test and normalized when necessary. Variables from different treatments were compared by ANOVA. Nuclear maturation from COCs cultured by 15 hours were considered as controls. Oocytes treated with FSK plus prorenin reached higher MI percentage (49.95%) than negative control (25.59%; P<0.05), however lower percentage than positive control (83.12% MI) and prorenin (78.34%) groups. Intra-oocyte cAMP concentrations were slightly reduced in COCs treated with prorenin plus FSK (8.66 ± 1.20) compared to negative control (10.33 ± 0.88). The positive control (4.00 ± 1.52) and prorenin (3.00 ± 0.57) groups showed lower concentration of cAMP compared to the negative control. Furthermore, concentration of cAMP (15.08±6.7, 9.91±3.68, 2.02±1.43 and 5.05±4.66) and cGMP (0.42±0.19, 0.31±0.07, 0.78±0.4 e 3.61±2.96) in cumulus cells and cGMP in oocytes (0.57±0.09, 0.71±0.09, 0.78±0.12 e 0.60±0.01) did not differ following the treatments (FSK, FSK plus prorenin, positive control and prorenin, respectively). In conclusion, our results indicate that prorenin acts on resumption of meiosis in bovine and suggest a regulation in the concentration of intra-oocyte cAMP." @default.
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• W2738864970 date "2015-03-01" @default.
• W2738864970 modified "2023-09-26" @default.
• W2738864970 title "Prorenin blocks forskolin effect and resumes meiosis in bovine oocytes" @default.
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