Homologous recombination and non-homologous end-joining repair pathways in bovine embryos with different developmental competence.
This study investigated the expression of genes controlling homologous recombination (HR), and non-homologous end-joining (NHEJ) DNA-repair pathways in bovine embryos of different developmental potential. It also evaluated whether bovine embryos can respond to DNA double-strand breaks (DSBs) induced with ultraviolet irradiation by regulating expression of genes involved in HR and NHEJ repair pathways. Embryos with high, intermediate or low developmental competence were selected based on the cleavage time after in vitro insemination and were removed from in vitro culture before (36 h), during (72 h) and after (96 h) the expected period of embryonic genome activation. All studied genes were expressed before, during and after the genome activation period regardless the developmental competence of the embryos. Higher mRNA expression of 53BP1 and RAD52 was found before genome activation in embryos with low developmental competence. Expression of 53BP1, RAD51 and KU70 was downregulated at 72 h and upregulated at 168 h post-insemination in response to DSBs induced by ultraviolet irradiation. In conclusion, important genes controlling HR and NHEJ DNA-repair pathways are expressed in bovine embryos, however genes participating in these pathways are only regulated after the period of embryo genome activation in response to ultraviolet-induced DSBs.
Universidade Federal de Santa Catarina, Campus Universitário de Curitibanos, Curitibanos, SC, Brazil.
Marcos
M
Henrique Barreta
Bernardo
B
Garziera Gasperin
Vitor
V
Braga Rissi
Matheus Pedrotti
MP
de Cesaro
Rogério
R
Ferreira
João Francisco
JF
de Oliveira
Paulo Bayard Dias
PB
Gonçalves
Vilceu
V
Bordignon
D000818:Animals; D034961:Antigens, Nuclear / Q000235:genetics / Q000378:metabolism; D002417:Cattle; D053903:DNA Breaks, Double-Stranded; D059766:DNA End-Joining Repair / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D047108:Embryonic Development / Q000502:physiology*; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental; D059765:Homologous Recombination; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D000072200:Ku Autoantigen; D051135:Rad51 Recombinase / Q000235:genetics / Q000378:metabolism; D051721:Rad52 DNA Repair and Recombination Protein / Q000235:genetics / Q000378:metabolism; D014466:Ultraviolet Rays
D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D051721:Rad52 DNA Repair and Recombination Protein; D051135:Rad51 Recombinase; D000072200:Ku Autoantigen