Abstract
Background/Purpose: Genomic imprinting is a specialized transcriptional mechanism that results in the unequal expression of alleles based on their parent-of-origin [1]. Many imprinted genes are critical for proper embryonic and fetaldevelopment [2] and disruption of genomic imprinting are associated with many development disorders [3]. Recently, increased frequencies of imprinting disorders have been correlated with the use of assisted reproductive technologies (ARTs)[2]. Rigorous and thorough testing of ARTs is required to determine their influence on genomic imprinting and development. I hypothesize that imprinting maintenance mechanisms are disrupted during early mouse development by the environmental insult of culture media used in human ARTs, and that loss of imprinting correlates with delayed embryonic development. Methods: The specific aims of my project are to develop a method to evaluate the methylation and expression patterns of 4 known imprinted genes in individual blastocysts. Results: We have successfully developed a novel method to evaluate both imprinted methylation and expression from a single mouse blastocyst. This method has been tested and results compared to methods used to evaluate imprinted methylation and expression separately; we have determined that results obtained with a combined protocol are equivalent to either alone. I will use this method to evaluate relationships between development rates in culture andgenomic imprinting, as well as the effects of various culture media used formouse and human embryo culture on genomic imprinting. Conclusion: This analysis allow for a more comprehensive study ofthe effects of environmental insult on genomic imprinting and preimplantation embryo development.
Published Version
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