Type

Journal Article

Authors

Patrick Lonergan
Maurice P Boland
Finian Martin
Catherine Moss
Dimitrios Rizos
Madeline Murphy
Trudee Fair

Subjects

Biochemistry

Topics
rna messenger female base sequence cattle animals genetics metabolism fertilization in vitro gene expression regulation developmental polymerase chain reaction male dna complementary embryonic development cleavage stage ovum

Analysis of differential maternal mRNA expression in developmentally competent and incompetent bovine two-cell embryos. (2003)

Abstract The main objective of this study was to identify mRNA transcripts associated with embryonic developmental competence. In cattle, mRNA transcripts, ribosomes, and proteins accumulated during the growth phase are drawn on to sustain maturation, fertilization, and the initial cell cycle divisions up to the 8- to 16-cell stage. Early cleaving mammalian zygotes are more likely to develop to the blastocyst stage than their later cleaving counterparts, thus reflecting the intrinsic quality of the oocytes from which they originated. We describe the combination of this well-established model for the retrospective determination of developmental competence in mammalian oocytes with a technique for wide screening of differential gene expression in different biological populations. Immature cumulus oocyte complexes were recovered from surface visible follicles on abattoir ovaries, washed, and submitted to routine in vitro maturation and fertilization. Two-cell embryos were removed from culture at 3-hr intervals from 24 to 42 hr post insemination (pi). Two populations of two-cell embryos were identified; those that cleaved early (before 27 hpi) and those that cleaved late (after 33 hpi). Suppressive subtractive hybridization was carried out on cDNA from the two populations, following which, differentially expressed amplicons were subcloned and sequenced. The sequences were submitted to the nonredundant and expressed sequence tag (EST) databases at NCBI using the BLAST algorithm. The differential expression of three selected candidate genes that were identified as putatively upregulated in the early cleaving zygotes were chosen for further investigations; histone H3, cyclin B1, and GDF-9B. Using quantitative real time PCR we have shown that histone H3A is significantly more abundant in embryos that cleave earliest.
Collections Ireland -> University College Dublin -> PubMed

Full list of authors on original publication

Patrick Lonergan, Maurice P Boland, Finian Martin, Catherine Moss, Dimitrios Rizos, Madeline Murphy, Trudee Fair

Experts in our system

1
P. Lonergan
University College Dublin
Total Publications: 190
 
2
M P Boland
University College Dublin
Total Publications: 103
 
3
Finian Martin
University College Dublin
Total Publications: 81
 
4
Catherine Moss
University College Dublin
Total Publications: 6
 
5
D Rizos
University College Dublin
Total Publications: 42
 
6
Madeline Murphy
University College Dublin
Total Publications: 37
 
7
T Fair
University College Dublin
Total Publications: 66