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Elongation and gene expression in bovine cloned embryos transferred to temporary recipients

Published online by Cambridge University Press:  08 June 2009

Lleretny Rodríguez-Alvarez
Affiliation:
Department of Animal Science, Faculty of Veterinary Medicine, Universidad de Concepción, Avenida Vicente Méndez 595, Chillán, Chile.
José Cox
Affiliation:
Department of Animal Science, Faculty of Veterinary Medicine, Universidad de Concepción, Avenida Vicente Méndez 595, Chillán, Chile.
Felipe Navarrete
Affiliation:
Department of Animal Science, Faculty of Veterinary Medicine, Universidad de Concepción, Avenida Vicente Méndez 595, Chillán, Chile.
Cristián Valdés
Affiliation:
Department of Animal Science, Faculty of Veterinary Medicine, Universidad de Concepción, Avenida Vicente Méndez 595, Chillán, Chile.
Teresa Zamorano
Affiliation:
Department of Animal Science, Faculty of Veterinary Medicine, Universidad de Concepción, Avenida Vicente Méndez 595, Chillán, Chile.
Ralf Einspanier
Affiliation:
Institute of Veterinary Biochemistry, Freie Universität Berlin, Oertzenweg 19b, D-14163 Berlin, Germany.
Fidel Ovidio Castro*
Affiliation:
Department of Animal Science, Faculty of Veterinary Medicine, Universidad de Concepción, Avenida Vicente Méndez 595, Chillán, Chile. Department of Animal Science, Faculty of Veterinary Medicine, Universidad de Concepción, Avenida Vicente Méndez 595, Chillán, Chile.
*
All correspondence to: Fidel Ovidio Castro. Department of Animal Science, Faculty of Veterinary Medicine, Universidad de Concepción, Avenida Vicente Méndez 595, Chillán, Chile. Tel: +56 42 207524. Fax: +56 42 207212. e-mail: fidcastro@udec.cl

Summary

Elongated embryos provide a unique source of information about trophoblastic differentiation, gene expression and maternal-embryonic interactions; however they are difficult and costly to obtain, especially elongated cloned embryos. One alternative is their production in heterologous temporary recipients such as sheep and goats. We aimed to produce elongated bovine cloned embryos using heterologous transfer to temporary recipients. Day-7 cloned cattle blastocysts were transferred to the uteri of ewes and goats and recovered as elongated structures at day 17. We evaluated elongation, length, presence of embryonic disc and expression of several important genes for embryonic development. We also produced homologous (cloned cattle embryos transferred into cattle uteri). Cloned bovine blastocysts were able to proceed with preimplantation development through elongation with high efficiency despite the species to which they were transferred. In qualitative and quantitative RT-PCR experiments we found differences in the pattern of gene expression among embryos recovered from different species. Sox2, Nanog and FGF-4 were markedly deregulated. No previous reports about the expression pattern of the studied genes had been published for elongated bovine cloned embryos produced in intermediate recipients, furthermore, the pattern of expression of Nanog, Oct4, Eomes, Cdx2, IFN-tau, Dicer, FGF-4 and Sox2 shown here are novel for elongated cloned bovine embryos created by hand-made cloning. Our data confirmed that sheep and goats can be used as temporary recipients. This model could serve as a basis for further research on gene expression and cellular changes during bovine peri-implantation development.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2009

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