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A retrospective model of oocyte competence: global mRNA and housekeeping transcripts are not associated with in vitro developmental outcome

Published online by Cambridge University Press:  27 April 2009

Fernando Henrique Biase ,
Lúcia Martelli ,
Giovana Krempel Fonseca Merighe ,
Weruska Karyna Freitas Santos Biase ,
Moyses Miranda ,
Lawrence Charles Smith  and
Flávio Vieira Meirelles
Fernando Henrique Biase*
Affiliation:
USP-FZEA – Departamento de Ciências Básicas, Rua Duque de Caxias Norte, 225, Pirassununga–SPBrazil. Departamento de Ciências Básicas, Faculdade de Zootecnia e Engenharia de Alimentos–Universidade de São Paulo, Pirassununga, São Paulo, Brasil. Departamento de Genética, Faculdade de Medicina de Ribeirão Preto–Universidade de São Paulo, Ribeirão Preto, São Paulo, Brasil.
Lúcia Martelli
Affiliation:
Departamento de Genética, Faculdade de Medicina de Ribeirão Preto–Universidade de São Paulo, Ribeirão Preto, São Paulo, Brasil.
Giovana Krempel Fonseca Merighe
Affiliation:
Departamento de Ciências Básicas, Faculdade de Zootecnia e Engenharia de Alimentos–Universidade de São Paulo, Pirassununga, São Paulo, Brasil.
Weruska Karyna Freitas Santos Biase
Affiliation:
Departamento de Ciências Básicas, Faculdade de Zootecnia e Engenharia de Alimentos–Universidade de São Paulo, Pirassununga, São Paulo, Brasil.
Moyses Miranda
Affiliation:
Departamento de Ciências Básicas, Faculdade de Zootecnia e Engenharia de Alimentos–Universidade de São Paulo, Pirassununga, São Paulo, Brasil.
Lawrence Charles Smith
Affiliation:
Departamento de Ciências Básicas, Faculdade de Zootecnia e Engenharia de Alimentos–Universidade de São Paulo, Pirassununga, São Paulo, Brasil. Centre de Recherche en Reproducion Animale, Faculté de Médicine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Quebec, Canada.
Flávio Vieira Meirelles
Affiliation:
USP-FZEA – Departamento de Ciências Básicas, Rua Duque de Caxias Norte, 225, Pirassununga–SPBrazil. Departamento de Ciências Básicas, Faculdade de Zootecnia e Engenharia de Alimentos–Universidade de São Paulo, Pirassununga, São Paulo, Brasil.
*
All correspondence to: Fernando H. Biase or Flávio Vieira Meirelles. USP-FZEA – Departamento de Ciências Básicas, Rua Duque de Caxias Norte, 225, Pirassununga–SPBrazil. Tel: +55 19 3565 4112. Fax: +55 19 3565 4117. e-mail: [email protected] or [email protected]
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Summary

Oocyte developmental competence depends on maternal stores that support development throughout a transcriptionally silent period during early embryogenesis. Previous attempts to investigate transcripts associated with oocyte competence have relied on prospective models, which are mostly based on morphological criteria. Using a retrospective model, we quantitatively compared mRNA among oocytes with different embryo development competence. A cytoplasm biopsy was removed from in vitro matured oocytes to perform comparative analysis of amounts of global polyadenylated (polyA) mRNA and housekeeping gene transcripts. After parthenogenetic activation of biopsied oocytes, presumptive zygotes were cultured individually in vitro and oocytes were classified according to embryo development: (i) blocked before the 8-cell stage; (ii) blocked between the 8-cell and morulae stages; or (iii) developed to the blastocyst stage. Sham-manipulated controls confirmed that biopsies did not alter development outcome. Total polyA mRNA amounts correlate with oocyte diameter but not with the ability to develop to the 8-cell and blastocyst stages. The last was also confirmed by relative quantification of GAPDH, H2A and Hprt1 transcripts. In conclusion, we describe a novel retrospective model to identify putative markers of development competence in single oocytes and demonstrate that global mRNA amounts at the metaphase II stage do not correlate with embryo development in vitro.

Keywords

BiopsyBovineEmbryoGene expression
Type
Research Article
Information
Zygote , Volume 17 , Issue 4 , November 2009 , pp. 289 - 295
Copyright
Copyright © Cambridge University Press 2009

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