Microarray analysis of gene expression during early development: a cautionary overview.
The rise of the 'omics' technologies started nearly a decade ago and, among them, transcriptomics has been used successfully to contrast gene expression in mammalian oocytes and early embryos. The scarcity of biological material that early developmental stages provide is the prime reason why the field of transcriptomics is becoming more and more popular with reproductive biologists. The potential to amplify scarce mRNA samples and generate the necessary amounts of starting material enables the relative measurement of RNA abundance of thousands of candidates simultaneously. So far, microarrays have been the most commonly used high-throughput method in this field. Microarray platforms can be found in a wide variety of formats, from cDNA collections to long or short oligo probe sets. These platforms generate large amounts of data that require the integration of comparative RNA abundance values in the physiological context of early development for their full benefit to be appreciated. Unfortunately, significant discrepancies between datasets suggest that direct comparison between studies is difficult and often not possible. We have investigated the sample-handling steps leading to the generation of microarray data produced from prehatching embryo samples and have identified key steps that significantly impact the downstream results. This review provides a discussion on the best methods for the preparation of samples from early embryos for microarray analysis and focuses on the challenges that impede dataset comparisons from different platforms and the reasons why methodological benchmarking performed using somatic cells may not apply to the atypical nature of prehatching development.
Robert
Claude
C
Laboratory of Functional Genomics of Early Embryonic Development, Laval University, Pavillon Comtois, Local 4221 Université Laval, Québec, Québec, Canada. claude.robert@fsaa.ulaval.ca
D000818:Animals; D001755:Blastocyst / Q000378:metabolism / Q000502:physiology; D004622:Embryo, Mammalian; D020869:Gene Expression Profiling / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D046228:Microarray Analysis / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D008954:Models, Biological; D009865:Oocytes / Q000378:metabolism / Q000502:physiology