Leemans R, Egger B, Loop T, Kammermeier L, He H, Hartmann B, Certa U, Hirth F, Reichert H
Institute of Zoology, Biocenter/Pharmacenter, University of Basel, CH-4056 Basel, Switzerland.
Proc Natl Acad Sci U S A. 2000 Oct 24;97(22):12138-43. doi: 10.1073/pnas.210066997.
Embryonic development in Drosophila is characterized by an early phase during which a cellular blastoderm is formed and gastrulation takes place, and by a later postgastrulation phase in which key morphogenetic processes such as segmentation and organogenesis occur. We have focused on this later phase in embryogenesis with the goal of obtaining a comprehensive analysis of the zygotic gene expression that occurs during development under normal and altered environmental conditions. For this, a functional genomic approach to embryogenesis has been developed that uses high-density oligonucleotide arrays for large-scale detection and quantification of gene expression. These oligonucleotide arrays were used for quantitative transcript imaging of embryonically expressed genes under standard conditions and in response to heat shock. In embryos raised under standard conditions, transcripts were detected for 37% of the 1,519 identified genes represented on the arrays, and highly reproducible quantification of gene expression was achieved in all cases. Analysis of differential gene expression after heat shock revealed substantial expression level changes for known heat-shock genes and identified numerous heat shock-inducible genes. These results demonstrate that high-density oligonucleotide arrays are sensitive, efficient, and quantitative instruments for the analysis of large scale gene expression in Drosophila embryos.
果蝇的胚胎发育具有两个阶段,早期会形成细胞胚盘并发生原肠胚形成,后期是原肠胚形成后的阶段,此阶段会发生诸如体节形成和器官发生等关键形态发生过程。我们专注于胚胎发育的后期阶段,目的是全面分析在正常和变化的环境条件下发育过程中合子基因的表达情况。为此,我们开发了一种用于胚胎发育的功能基因组学方法,该方法使用高密度寡核苷酸阵列对基因表达进行大规模检测和定量分析。这些寡核苷酸阵列用于在标准条件下以及热休克反应中对胚胎表达基因进行定量转录本成像。在标准条件下饲养的胚胎中,检测到阵列上所代表的1519个已鉴定基因中的37%有转录本,并且在所有情况下都实现了基因表达的高度可重复定量。热休克后差异基因表达分析揭示了已知热休克基因的大量表达水平变化,并鉴定出许多热休克诱导基因。这些结果表明,高密度寡核苷酸阵列是用于分析果蝇胚胎大规模基因表达的灵敏、高效且定量的工具。