| Autor: |
Costa CB; Graduate Program in Pharmacology and Biotechnology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu 18618-970, SP, Brazil.; Department of Biological Sciences, School of Sciences, Humanities and Languages, São Paulo State University (UNESP), Assis 19806-900, SP, Brazil.; Laboratory of Animal Reproduction, Department of Veterinary Medicine, University of Londrina (UEL), Londrina 86057-970, PR, Brazil., Silva NCD; Laboratory of Animal Reproduction, Department of Veterinary Medicine, University of Londrina (UEL), Londrina 86057-970, PR, Brazil., Silva AN; Graduate Program in Anatomy of Domestic and Wild Animals, University of São Paulo (USP), Pirassununga 13635-000, SP, Brazil.; Department of Veterinary Medicine, College of Animal Science and Food Engineering, University of São Paulo (USP), Pirassununga 13635-000, SP, Brazil., Pioltine EM; Department of Biological Sciences, School of Sciences, Humanities and Languages, São Paulo State University (UNESP), Assis 19806-900, SP, Brazil., Dellaqua TT; Graduate Program in Pharmacology and Biotechnology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu 18618-970, SP, Brazil., Zangirolamo AF; Laboratory of Animal Reproduction, Department of Veterinary Medicine, University of Londrina (UEL), Londrina 86057-970, PR, Brazil.; National Institute of Science and Technology for Dairy Production Chain (INCT-LEITE), University of Londrina (UEL), Londrina 86057-970, PR, Brazil., Meirelles FV; Department of Veterinary Medicine, College of Animal Science and Food Engineering, University of São Paulo (USP), Pirassununga 13635-000, SP, Brazil., Seneda MM; Laboratory of Animal Reproduction, Department of Veterinary Medicine, University of Londrina (UEL), Londrina 86057-970, PR, Brazil.; National Institute of Science and Technology for Dairy Production Chain (INCT-LEITE), University of Londrina (UEL), Londrina 86057-970, PR, Brazil., Nogueira MFG; Graduate Program in Pharmacology and Biotechnology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu 18618-970, SP, Brazil.; Department of Biological Sciences, School of Sciences, Humanities and Languages, São Paulo State University (UNESP), Assis 19806-900, SP, Brazil. |
| Abstrakt: |
The use of C-type natriuretic peptide (CNP) in the interaction with the oocyte and in the temporary postponement of spontaneous meiosis resumption has already been well described. However, its action in pre-implantation developmental-stage embryos is yet to be understood. Thus, our study aimed to detect the presence of the canonical CNP receptor (natriuretic peptide receptor, NPR2) in germinal vesicle (GV)-, metaphase II (MII)-, presumptive zygote (PZ)-, morula (MO)-, and blastocyst (BL)-stage embryos and, later, to observe possible modulations on the embryos when co-cultured with CNP. In Experiment I, we detected and quantified NPR2 on the abovementioned embryo stages. Further, in Experiment II, we intended to test different concentrations (100, 200, or 400 nM of CNP) at different times of inclusion in the in vitro culture (IVC; inclusion from the beginning, i.e., day 1, or from day 5). In Experiment III, 400 nM of CNP was used on day 1 (D1) in the IVC, which was not demonstrated to be embryotoxic, and it showed potentially promising results in the blastocyst production rate when compared to the control. Thus, we analyzed the embryonic development rates of bovine embryos (D7) and hatching kinetics (D7, D8, and D9). Subsequently, morula and blastocyst were collected and evaluated for transcript abundance of their competence and quality (apoptosis, oxidative stress, proliferation, and differentiation) and lipid metabolism. Differences with probabilities less than p < 0.05, and/or fold change (FC) > 1.5, were considered significant. We demonstrate the presence of NPR2 until the blastocyst development stage, when there was a significant decrease in membrane receptors. There was no statistical difference in the production rate after co-culture with 400 nM CNP. However, when we evaluated the abundance of morula transcripts, there was an upregulated transcription in ADCY6 ( p = 0.057) and downregulated transcripts in BMP15 ( p = 0.013), ACAT1 ( p = 0.040), and CASP3 ( p = 0.082). In addition, there was a total of 12 transcriptions in morula that presented variation FC > 1.5. In blastocysts, the treatment with CNP induced upregulation in BID , CASP3 , SOX2 , and HSPA5 transcripts and downregulation in BDNF , NLRP5 , ELOVL1 , ELOVL4 , IGFBP4 , and FDX1 transcripts (FC > 1.5). Thus, our study identified and quantified the presence of NPR2 in bovine pre-implantation embryos. Furthermore, 400 nM of CNP in IVC, a concentration not previously described in the literature, modulated some transcripts related to embryonic metabolism, and this was not embryotoxic morphologically. |
