Effects of in vitro fertilization and embryo culture on TRP53 and Bax expression in B6 mouse embryos
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BioMed Central
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Effects of in vitro fertilization and embryo culture on TRP53 and Bax expression in B6 mouse embryos Vashe Chandrakanthan1, Aiqing Li1, Omar Chami1 and Christopher O'Neill*1,2 Address: 1Discipline of Physiology, University of Sydney, Royal North Shore Hospital, St Leonards, NSW 2065, Australia and 2Discipline of Medicine, University of Sydney, Royal North Shore Hospital, St Leonards, NSW 2065, Australia Email: Vashe Chandrakanthan - [email protected]; Aiqing Li - [email protected]; Omar Chami - [email protected]; Christopher O'Neill* - [email protected] * Corresponding author
Published: 21 November 2006 Reproductive Biology and Endocrinology 2006, 4:61
doi:10.1186/1477-7827-4-61
Received: 08 September 2006 Accepted: 21 November 2006
This article is available from: http://www.rbej.com/content/4/1/61 © 2006 Chandrakanthan et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract In the mouse, embryo culture results in a characteristic phenotype of retarded embryo preimplantation development and reduced numbers of cells within embryos. The expression of TRP53 is central to the regulation of the cell's capacity to proliferate and survive. In this study we found that Trp53 mRNA is expressed throughout the preimplantation stage of development. Levels of TRP53 protein expression were low during the cleavage stages and increased at the morula and blastocyst stages in B6 embryos collected from the reproductive tract. Embryos collected at the zygote stage and cultured for 96 h also showed low levels of TRP53 expression at precompaction stages. There were higher levels of TRP53 in cultured morula and the level in cultured blastocysts was clearly increased above blastocysts collected directly from the uterus. Immunolocalization of TRP53 showed that its increased expression in cultured blastocysts corresponded with a marked accumulation of TRP53 within the nuclei of embryonic cells. This pattern of expression was enhanced in embryos produced by in vitro fertilization and subjected to culture. The TRP53 was transcriptionally active since culture also induced increased expression of Bax, yet this did not occur in embryos lacking Trp53 (Trp53-/-). The rate of development of Trp53-/- zygotes to the blastocyst stage was not different to wildtype controls when embryos were cultured in groups of ten but was significantly faster when cultured individually. The results show that zygote culture resulted in the accumulation of transcription activity of TRP53 in the resulting blastocysts. This accounts for the adverse effects of culture of embryos individually, but does not appear to be the sole cause of the retarded preimplantation stage growth phenotype associated with culture in vitro.
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