Identification of differentially expressed genes in individual bovine preimplantation embryos produced by nuclear transfer: improper reprogramming of genes required for development.
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, cytokeratin 19, and vimentin), three metabolic genes (phosphoribosyl pyrophosphate synthetase 1, mitochondrial acetoacetyl-coenzyme A thiolase, and alpha-glucosidase), two lysosomal-related genes (prosaposin and lysosomal-associated membrane protein 2), and a gene associated with stress responses (heat shock protein 27) along with major histocompatibility complex class I, nidogen 2, a putative transport protein, heterogeneous 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 it uses a rigorous experimental design that emphasizes statistical significance to identify differentially expressed genes between NT and IVF embryos in cattle.
Pfister-Genskow
Martha
M
Infigen, Inc., DeForest, Wisconsin 53532, USA.
Myers
Cena
C
Childs
Lynette A
LA
Lacson
Jenine C
JC
Patterson
Thomas
T
Betthauser
Jeffery M
JM
Goueleke
Paul J
PJ
Koppang
Richard W
RW
Lange
Gail
G
Fisher
Patricia
P
Watt
Steven R
SR
Forsberg
Erik J
EJ
Zheng
Ying
Y
Leno
Gregory H
GH
Schultz
Richard M
RM
Liu
Bing
B
Chetia
Chiranjeet
C
Yang
Xiao
X
Hoeschele
Ina
I
Eilertsen
Kenneth J
KJ
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism*; D002417:Cattle / Q000196:embryology* / Q000235:genetics / Q000378:metabolism; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism*; D019976:Cloning, Organism / Q000379:methods; D003627:Data Interpretation, Statistical; D047108:Embryonic Development / Q000235:genetics / Q000502:physiology*; D005307:Fertilization in Vitro / Q000662:veterinary; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000235:genetics / Q000502:physiology*; D016014:Linear Models; D053652:Nuclear Transfer Techniques; D020411:Oligonucleotide Array Sequence Analysis