Transcriptional profiling of pig embryogenesis by using a 15-K member unigene set specific for pig reproductive tissues and embryos.
Differential mRNA expression patterns were evaluated between germinal vesicle oocytes (pgvo), four-cell (p4civv), blastocyst (pblivv), and in vitro-produced four-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, pgvo 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. There were 1409 and 1696 differentially detected cDNAs between the in vitro- and in vivo-produced embryos at the four-cell and blastocyst stages, respectively, without the Benjamini and Hochberg false-discovery-rate multiple correction test. Real-time polymerase chain reaction (PCR) analysis on four genes at the four-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.
Whitworth
K M
KM
Department of Animal Science, University of Missouri-Columbia, Missouri 65211, USA.
Agca
C
C
Kim
J-G
JG
Patel
R V
RV
Springer
G K
GK
Bivens
N J
NJ
Forrester
L J
LJ
Mathialagan
N
N
Green
J A
JA
Prather
R S
RS
D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D018076:DNA, Complementary / Q000302:isolation & purification; D047108:Embryonic Development / Q000235:genetics; D005260:Female; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D009865:Oocytes / Q000502:physiology*; D011247:Pregnancy; D012313:RNA / Q000592:standards; D012015:Reference Standards; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine / Q000235:genetics*; D014158:Transcription, Genetic
D018076:DNA, Complementary; D012313:RNA