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Supplementation with the histone deacetylase inhibitor trichostatin A during in vitro culture of bovine embryos

Published online by Cambridge University Press:  26 August 2011

Clara Slade Oliveira*
Affiliation:
UNESP–Via de Acesso Professor Paulo Donato Castellane, CEP 14884–900, Jaboticabal, São Paulo, Brazil.
Naiara Zoccal Saraiva
Affiliation:
Department of Preventive Veterinary Medicine and Animal Reproduction, Faculty of Agricultural and Veterinary Sciences, Sao Paulo State University, Access Road Professor Paulo Donato Castellane, zip code 14884–900, Jaboticabal, Brazil.
Marcela Maria de Souza
Affiliation:
Department of Preventive Veterinary Medicine and Animal Reproduction, Faculty of Agricultural and Veterinary Sciences, Sao Paulo State University, Access Road Professor Paulo Donato Castellane, zip code 14884–900, Jaboticabal, Brazil.
Tatiane de Almeida Drummond Tetzner
Affiliation:
Department of Preventive Veterinary Medicine and Animal Reproduction, Faculty of Agricultural and Veterinary Sciences, Sao Paulo State University, Access Road Professor Paulo Donato Castellane, zip code 14884–900, Jaboticabal, Brazil.
Marina Ragagnin de Lima
Affiliation:
Department of Preventive Veterinary Medicine and Animal Reproduction, Faculty of Agricultural and Veterinary Sciences, Sao Paulo State University, Access Road Professor Paulo Donato Castellane, zip code 14884–900, Jaboticabal, Brazil.
Joaquim Mansano Garcia
Affiliation:
Department of Preventive Veterinary Medicine and Animal Reproduction, Faculty of Agricultural and Veterinary Sciences, Sao Paulo State University, Access Road Professor Paulo Donato Castellane, zip code 14884–900, Jaboticabal, Brazil.
*
All correspondence to: Clara Slade Oliveira. UNESP–Via de Acesso Professor Paulo Donato Castellane, CEP 14884–900, Jaboticabal, São Paulo, Brazil. Tel: +55 16 32092633/+55 16 97242262. Fax: +55 21 25258804. E-mail: [email protected]

Summary

Trichostatin A (TSA) is a histone deacetylase inhibitor that induces histone hyperacetylation and increases gene expression levels. The aim of the present study was to establish a suitable condition for the use of TSA in in vitro cultures of bovine embryos, and to determine whether TSA would increase blastocyst rates by improvement of chromatin remodelling during embryonic genome activation and by increasing the expression of crucial genes during early development. To test this hypothesis, 8-cell embryos were exposed to four concentrations of TSA for different periods of time to establish adequate protocols. In a second experiment, three experimental groups were selected for the evaluation of embryo quality based on the following parameters: apoptosis, total cell number and blastocyst hatching. TSA promoted embryonic arrest and degeneration at concentrations of 15, 25 and 50 nM. All treated groups presented lower blastocyst rates. Exposure of embryos to 5 nM for 144 h and to 15 nM for 48 h decreased blastocyst hatching. However, the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay (TUNEL) assay revealed similar apoptosis rates and total cell numbers in all groups studied. Although, in the present study, TSA treatment did not improve the parameters studied, the results provided background information on TSA supplementation during in vitro culture of bovine embryos and showed that embryo quality was apparently not affected, despite a decrease in blastocyst rate after exposure to TSA.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2011

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