Analyses of apoptosis and DNA damage in bovine cumulus cells after in vitro maturation with different copper concentrations: consequences on early embryo development.
The aim of this study was to investigate the influence of copper (Cu) during in vitro maturation (IVM) on apoptosis and DNA integrity of cumulus cells (CC); and oocyte viability. Also, the role of CC in the transport of Cu during IVM was evaluated on oocyte developmental capacity. Damage of DNA was higher in CC matured without Cu (0 µg/dl Cu, P < 0.01) with respect to cells treated with Cu for cumulus-oocyte complexes (COCs) exposed to 0, 20, 40, or 60 µg/dl Cu). The percentage of apoptotic cells was higher in CC matured without Cu than in CC matured with Cu. Cumulus expansion and viability of CC did not show differences in COC treated with 0, 20, 40, or 60 µg/dl Cu during IVM. After in vitro fertilization (IVF), cleavage rates were higher in COC and DO + CC (denuded oocytes + CC) with or without Cu than in DO. Independently of CC presence (COC, DO + CC or DO) the blastocyst rates were higher when 60 µg/dl Cu was added to IVM medium compared to medium alone. These results indicate that Cu supplementation to IVM medium: (i) decreased DNA damage and apoptosis in CC; (ii) did not modify oocyte viability and cumulus expansion; and (iii) improved subsequent embryo development up to blastocyst stage regardless of CC presence during IVM.
Cátedra de Fisiología,Laboratorio de Nutrición Mineral,Facultad de Ciencias Veterinarias,Universidad Nacional de La Plata,calle 60 y 118 s/n,CP (1900),La Plata,Buenos Aires,Argentina.
D E
DE
Rosa
Instituto de Genética Veterinaria Prof. Fernando N. Dulout (IGEVET),Facultad de Ciencias Veterinarias,Universidad Nacional de La Plata - CONICET,calle 60 y 118 s/n,CP (1900),La Plata,Buenos Aires,Argentina.
J M
JM
Anchordoquy
Instituto de Genética Veterinaria Prof. Fernando N. Dulout (IGEVET),Facultad de Ciencias Veterinarias,Universidad Nacional de La Plata - CONICET,calle 60 y 118 s/n,CP (1900),La Plata,Buenos Aires,Argentina.
J P
JP
Anchordoquy
Instituto de Genética Veterinaria Prof. Fernando N. Dulout (IGEVET),Facultad de Ciencias Veterinarias,Universidad Nacional de La Plata - CONICET,calle 60 y 118 s/n,CP (1900),La Plata,Buenos Aires,Argentina.
M A
MA
Sirini
Instituto de Genética Veterinaria Prof. Fernando N. Dulout (IGEVET),Facultad de Ciencias Veterinarias,Universidad Nacional de La Plata - CONICET,calle 60 y 118 s/n,CP (1900),La Plata,Buenos Aires,Argentina.
J A
JA
Testa
Cátedra de Fisiología,Laboratorio de Nutrición Mineral,Facultad de Ciencias Veterinarias,Universidad Nacional de La Plata,calle 60 y 118 s/n,CP (1900),La Plata,Buenos Aires,Argentina.
G A
GA
Mattioli
Instituto de Genética Veterinaria Prof. Fernando N. Dulout (IGEVET),Facultad de Ciencias Veterinarias,Universidad Nacional de La Plata - CONICET,calle 60 y 118 s/n,CP (1900),La Plata,Buenos Aires,Argentina.
C C
CC
Furnus
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001755:Blastocyst / Q000166:cytology / Q000502:physiology; D002417:Cattle; D002478:Cells, Cultured; D003300:Copper / Q000008:administration & dosage / Q000494:pharmacology*; D054885:Cumulus Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D004249:DNA Damage / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D005260:Female; D005307:Fertilization in Vitro; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods*; D008297:Male; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology
D003300:Copper
Apoptosis; DNA damage; Early development; Minerals