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BOR - Papers in Press, published online ahead of print February 9, 2005.
Biol Reprod 2005, 10.1095/biolreprod.104.037952
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Submitted November 22, 2004
Returned for revision December 6, 2004
Accepted February 7, 2005

Reproductive Technology


Transcriptional Profiling of Pig Embryogenesis by Using a 15K Member Unigene Set Specific for Pig Reproductive Tissues and Embryos

K. M. Whitworth , C. Agca , J.- G. Kim , R. V. Patel , G. K. Springer , N. Bivens , L. J. Forrester , N. Mathialagan , J. A. Green , and R. S. Prather *

* To whom correspondence should be addressed. E-mail: pratherr{at}missouri.edu.

Abstract
Differential mRNA expression patterns were evaluated between germinal vesicle oocytes (pgvo), 4-cell (p4civv), blastocyst (pblivv) and in vitro produced 4-cell (p4civp) and in vitro produced blastocyst (pblivp) stage embryos to determine key transcripts responsible for early embryonic development in the pig. Five comparisons were made: pgvo to p4civv, p4civv to pblivv, p4civv to p4civp, and pblivv to pblivp. ANOVA (p<0.05) was performed with the Benjamini and Hochberg False Discovery Rate multiple correction test on each comparison. A comparison of pgvo to p4civv, p4civv to pblivv, and pgvo to pblivv resulted in 3214, 1989 and 4528 differentially detected cDNAs, respectively. Real-time PCR analysis on seven transcripts showed an identical pattern of changes in expression as observed on the microarrays while one transcript deviated at a single cell stage. The in vitro and in vivo comparisons were conducted without the Benjamini and Hochberg False Discovery Rate multiple correction test. There were 1409 and 1696 differentially detected cDNAs between the in vitro and in vivo produced embryos at the 4-cell and blastocyst stages, respectively, without the Benjamini and Hochberg False Discovery Rate multiple correction test. Real-time PCR analysis on four genes at the 4-cell stage showed an identical pattern of gene expression as found on the microarrays. Real-time PCR analysis on four of five genes at the blastocyst stage showed an identical pattern of gene expression as found on the microarrays. Thus only 1 of the 39 comparisons of the pattern of gene expression exhibited a major deviation between the microarray and the real-time PCR. These results illustrate the complex mechanisms involved in pig early embryonic development.

Key words: Early development • Gene regulation • In vitro fertilization


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