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BOR - Papers in Press, published online ahead of print October 13, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.031799
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Submitted May 25, 2004
Returned for revision June 18, 2004
Accepted September 21, 2004

Embryo


Identification of Differentially Expressed Genes in Individual Bovine Preimplantation Embryos Produced by Nuclear Transfer: Improper Reprogramming of Genes Required for Development

Martha Pfister-Genskow , Cena Myers , Lynette A. Childs , Jenine C. Lacson , Thomas Patterson , Jeffery M. Betthauser , Paul J. Goueleke , Richard W. Koppang , Gail Lange , Patricia Fisher , Steven R. Watt , Erik J. Forsberg , Ying Zheng , Gregory H. Leno , Richard M. Schultz , Bing Liu , Chiranjeet Chetia , Xiao Yang , Ina Hoeschele , and Kenneth J. Eilertsen *

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

Abstract
Using an interwoven loop experimental design in conjunction with highly conservative Linear Mixed Model Methodology using estimated variance components, 18 genes differentially expressed between nuclear transfer- (NT) and in vitro fertilization- (IVF) produced embryos were identified. The set is comprised of three intermediate filament protein genes (cytokeratin 8, 19, and vimentin), three metabolic genes (phosphoribosyl pyrophosphate synthetase 1, mitochondrial acetoacetyl-CoA thiolase and alpha-glucosidase), two lysosomal related genes (prosaposin and lysosomal-associated membrane protein 2), and a gene associated with stress responses (Hsp27), along with major histocompatibility complex class I, nidogen 2, a putative transport protein, heterogenous nuclear ribonuclear protein K, mitochondrial 16S rRNA, and ES1 (a zebrafish orthologue of unknown function). The three remaining genes are novel. To our knowledge, this is the first report comparing individual embryos produced by NT and IVF using cDNA microarray technology for any species, and uses a rigorous experimental design that emphasizes statistical significance to identify differentially expressed genes between NT and IVF embryos in cattle.

Key words: Embryo • Developmental biology • Early development • Trophoblast


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