Beta-hydroxybutyrate alters bovine preimplantation embryo development through transcriptional and epigenetic mechanisms†

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Sangalli, Juliano Rodrigues ; Nociti, Ricardo Perecin ; Chiaratti, Marcos Roberto ; Bridi, Alessandra ; Botigelli, Ramon Cesar ; Ambrizi, Dewison Ricardo ; Saraiva, Helena Fabiana Reis de Almeida ; Perecin, Felipe ; da Silveira, Juliano Coelho ; Ross, Pablo Juan ; Meirelles, Flavio Vieira

• Document type: Journal article
• Source: BIOLOGY OF REPRODUCTION; v. 112, n. 2, p. 20-pg., 2024-12-12.

Abstract

Developing embryos are susceptible to fluctuations in the nutrients and metabolites concentrations within the reproductive tract, which can lead to alterations in their developmental trajectory. Ketotic dairy cows have diminished fertility, and elevated levels of the ketone body beta-hydroxybutyrate (BHB) have been associated with poor embryonic development. We used an in vitro model based on either in vitro fertilization (IVF) or parthenogenesis to investigate the effects of BHB on the preimplantation bovine embryo development, epigenome, and transcriptome. Embryo culture medium was supplemented with BHB at a similar concentration to that present in the blood of cows suffering with severe ketosis, followed by analysis of blastocysts formation rate, diameter, total number of cells, levels of H3K9 beta-hydroxybutyrylation (H3K9bhb), apoptosis, and transcriptional alterations. As a result, we observed that BHB reduced the blastocysts rates, the diameter and the total number of cells in both parthenotes and IVF embryos. Exposure to BHB for either 3 or 7 days greatly increased the H3K9bhb levels in parthenotes at the 8-cells and blastocyst stages, and affected the expression of HDAC1, TET1, DNMT1, KDM6B, NANOG, and MTHFD2 genes. Additionally, culture of IVF embryos with BHB for 7 days dramatically increased H3K9bhb and reduced NANOG in blastocysts. RNA-seq analysis of IVF blastocysts revealed that BHB modulated the expression of 118 genes, which were involved with biological processes such as embryonic development, implantation, reproduction, proliferation, and metabolism. These findings provided valuable insights into the mechanisms through which BHB disrupts preimplantation embryonic development and affects the fertility in dairy cows. Beta-hydroxybutyrate induces histone lysine beta-hydroxybutyrylation in parthenotes and IVF-embryos, decrease their development to blastocyst, and alters the transcription of genes involved in implantation, reproduction, proliferation, and metabolism. (AU)

• FAPESP's process: 16/13416-9 - Use of the ketone body beta-hydroxybutyric acid as an experimental tool to study the link among metabolism, epigenetic and reproduction in cattle
• Grantee: Juliano Rodrigues Sangalli
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 22/01433-7 - Multi-user equipment approved in the grant FAPESP 21/06645-0: Inverted Microscope Yhunder Imager 3D Assay
• Grantee: Juliano Coelho da Silveira
• Support Opportunities: Multi-user Equipment Program
• FAPESP's process: 22/06581-4 - Dietary nutrients and metabolites influencing the bovine epigenome, the reproduction, the health and the food production
• Grantee: Juliano Rodrigues Sangalli
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 23/13753-9 - Dietary nutrients and metabolites influencing the bovine epigenome, the reproduction, the health and the food production.
• Grantee: Juliano Rodrigues Sangalli
• Support Opportunities: Scholarships in Brazil - Young Researchers
• FAPESP's process: 13/08135-2 - CTC - Center for Cell-Based Therapy
• Grantee: Dimas Tadeu Covas
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 18/09552-0 - Epigenetic effect of the ketone body beta-hydroxybutyrate on histone beta-hydroxybutyrylation in bovine cumulus-oocyte complexes
• Grantee: Juliano Rodrigues Sangalli
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor