Biol Reprod
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BOR - Papers in Press, published online ahead of print October 17, 2002.
Biol Reprod 2002, 10.1095/biolreprod.102.005991
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BIOLOGY OF REPRODUCTION 68, 383–389 (2003)
DOI: 10.1095/biolreprod.102.005991
© 2003 by the Society for the Study of Reproduction, Inc.

Embryo

Quantification of Histone Acetyltransferase and Histone Deacetylase Transcripts During Early Bovine Embryo Development1

Serge McGrawa, Claude Roberta, Lyne Massicottea, and Marc-André Sirard2,a

a Centre de Recherche en Biologie de la Reproduction, Department of Animal Sciences, Laval University, Québec, Canada G1K 7P4

Mammalian oocytes are very unique cells with an unlimited developmental potential. These totipotent cells are able to remove existing gene-expression patterns and to impose new ones. However, genome reprogramming is still a mystery. Posttranslational modifications by acetylation of the N-termini portion of histones composing the nucleosome are involved in genome reprogramming. These modifications alter the higher-order chromatin structure to render the DNA accessible to the regulatory and transcriptional machinery. In the present study, we have investigated, to our knowledge for the first time, precise expression patterns of seven genes involved in chromatin structure throughout bovine embryo development. Oocytes harvested from bovine ovaries were used for in vitro production of germinal vesicle oocytes, metaphase II oocytes, 2- and 8-cell embryos, and blastocysts. Total RNA was extracted from pools (triplicates) of 20 oocytes or from embryos of each developmental stage. By means of quantitative reverse transcription-polymerase chain reaction using SYBR Green to detect double-stranded DNA, mRNA expression profiles for histone deacetylases (HDAC1, HDAC2, HDAC3, and HDAC7), histone acetyltransferases (GCN5 and HAT1), and histone H2A were established. Transcripts for all genes were detected at all stages from the oocyte to the blastocyst. The HDAC1, HDAC2 (class I HDAC), and HAT1 (type B HAT) revealed similar expression profiles. The HDAC3 (class I HDAC) tends to have an expression profile similar to those of HDAC1, HDAC2, and HAT1, whereas the HDAC7 (class II HDAC) and GCN5 (type A HAT) profiles were different from those three. These results indicate variable levels of histone deacetylases and histone acetyltransferases throughout embryonic development and may indicate the ones that are involved in somatic remodeling.

1 Supported by the Canadian National Sciences and Engineering Research Council.

2 Correspondence. FAX: 418 656 3766; marc-andre.sirard{at}crbr.ulaval.ca






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Copyright © 2003 by the Society for the Study of Reproduction.