Catalytic inhibition of H3K9me2 writers disturbs epigenetic marks during bovine nuclear reprogramming

Author(s):Show less -

Sampaio, Rafael Vilar ^{[1, 2, 3]} ; Sangalli, Juliano Rodrigues ^{[1, 2]} ; Camara De Bem, Tiago Henrique ^[1] ; Ambrizi, Dewison Ricardo ^[1] ; del Collado, Maite ^[1] ; Bridi, Alessandra ^[1] ; Cavalcante Mendes de Avila, Ana Clara Faquineli ^[1] ; Macabelli, Carolina Habermann ^[4] ; Oliveira, Lilian de Jesus ^[1] ; da Silveira, Juliano Coelho ^[1] ; Chiaratti, Marcos Roberto ^[4] ; Perecin, Felipe ^[1] ; Bressan, Fabiana Fernandes ^[1] ; Smith, Lawrence Charles ^[3] ; Ross, Pablo J. ^[2] ; Meirelles, Flavio Vieira ^[1]

Affiliation

^[1] Univ Sao Paulo, Fac Zootecnia & Engn Alimentos, Dept Med Vet, Pirassununga, SP - Brazil

^[2] Univ Calif Davis, Dept Anim Sci, Davis, CA 95616 - USA

^[3] Univ Montreal, Fac Vet Med, Ctr Rech Reprod & Fertilite, St Hyacinthe, PQ - Canada

^[4] Univ Fed Sao Carlos, Dept Genet & Evolucao, Sao Carlos, SP - Brazil

• Document type: Journal article
• Source: SCIENTIFIC REPORTS; v. 10, n. 1 JUL 13 2020.
• Web of Science Citations: 0

Abstract

Orchestrated events, including extensive changes in epigenetic marks, allow a somatic nucleus to become totipotent after transfer into an oocyte, a process termed nuclear reprogramming. Recently, several strategies have been applied in order to improve reprogramming efficiency, mainly focused on removing repressive epigenetic marks such as histone methylation from the somatic nucleus. Herein we used the specific and non-toxic chemical probe UNC0638 to inhibit the catalytic activity of the histone methyltransferases EHMT1 and EHMT2. Either the donor cell (before reconstruction) or the early embryo was exposed to the probe to assess its effect on developmental rates and epigenetic marks. First, we showed that the treatment of bovine fibroblasts with UNC0638 did mitigate the levels of H3K9me2. Moreover, H3K9me2 levels were decreased in cloned embryos regardless of treating either donor cells or early embryos with UNC0638. Additional epigenetic marks such as H3K9me3, 5mC, and 5hmC were also affected by the UNC0638 treatment. Therefore, the use of UNC0638 did diminish the levels of H3K9me2 and H3K9me3 in SCNT-derived blastocysts, but this was unable to improve their preimplantation development. These results indicate that the specific reduction of H3K9me2 by inhibiting EHMT1/2 during nuclear reprogramming impacts the levels of H3K9me3, 5mC, and 5hmC in preimplantation bovine embryos. (AU)

• FAPESP's process: 15/08807-6 - H3K9 methylation as regulator of epigenetic memory on bovine nuclear reprograming.
• Grantee: Rafael Vilar Sampaio
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/22887-0 - Cell-secreted vesicles containing miRNAs modulate epigenetic changes during in vitro culture of bovine gametes and embryos
• Grantee: Juliano Coelho da Silveira
• Support Opportunities: Research Grants - Young Investigators Grants
• 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: 14/50947-7 - INCT 2014: in Stem Cell and Cell Therapy
• Grantee: Dimas Tadeu Covas
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 14/21034-3 - Intra and intercellular routes during foliculogenesis and in vivo and in vitro oocyte maturation
• Grantee: Felipe Perecin
• Support Opportunities: Regular Research Grants
• FAPESP's process: 15/25111-5 - Analysis and modulation of H3K9 methylation in bovine somatic cells
• Grantee: Rafael Vilar Sampaio
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• FAPESP's process: 12/50533-2 - GIFT: genomic improvement of fertilization traits in Danish and Brazilian cattle
• Grantee: Marcelo Fábio Gouveia Nogueira
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 13/07160-3 - Influence of the 5-hydroxymethylcytosine on Nuclear Reprogramming of Bovines
• Grantee: Rafael Vilar Sampaio
• Support Opportunities: Scholarships in Brazil - Doctorate