Potential and limitations of bovine-specific arrays for the analysis of mRNA levels in early development: preliminary analysis using a bovine embryonic array.
New insights into the early development of large mammals are becoming available through the measurement of differential mRNA levels in oocytes and preimplantation embryos. These advances in knowledge are rapidly picking up in pace, mainly owing to the advantages brought by new molecular biology approaches being developed. The possibility of amplifying the starting material and therefore making measurements in single embryo units is now feasible. With these tools, the evaluation of variations in gene expression patterns during the preimplantation period or the impact of culture on mRNA levels is now possible. However, it is important to keep in mind that these methods still have limitations associated with sample preparation or the use of the appropriate controls. Even proper methods of analysis are very important to achieve the full benefit of the application of these tools. The present paper describes some of the potential, as well as limitations, of mRNA level analysis in early embryos, especially for microarray analysis. We have generated a bovine cDNA array (>2000 clones) that contains expressed sequence tags (ESTs) collected from various preimplantation development stages. Using this chip, we have initiated the characterisation of global mRNA level patterns of several key developmental stages from the immature oocyte to the blastocyst stage. As expected, the hybridisation results indicate very different expression profiles involving hundreds of genes when comparing oocyte and blastocyst samples to a reference mRNA sample made from a pool of ESTs from pooled somatic tissues. Although this array is still in its preliminary stage and the EST bank has not been processed to contain only unigenes, it is already a very useful tool for discovering candidate genes that may play important roles during early embryonic life.
Sirard
Marc-André
MA
Centre de Recherche en Biologie de la Reproduction, Dtement des Sciences Animales, Universital, Sainte-Foy, Quebec G1K 7P4, Canada. marc-andre.sirard@crbr.ulaval.ca
Dufort
Isabelle
I
Vallée
Maud
M
Massicotte
Lyne
L
Gravel
Catherine
C
Reghenas
Hélène
H
Watson
Andrew J
AJ
King
W Allan
WA
Robert
Claude
C
D000818:Animals; D002417:Cattle / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics; D046149:Embryo Culture Techniques / Q000662:veterinary; D047108:Embryonic Development / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000662:veterinary*; D012333:RNA, Messenger / Q000032:analysis*; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D012333:RNA, Messenger