Gene expression profiling in embryonic mouse lenses

[摘要] Purpose: In this study, we used laser capture microdissection (LCM)and microarray hybridization technology to compare the gene expressionprofiles of mouse embryonic days 10 and 12 lenses (E10 and E12).Methods: Lens cells of C57/BL6 mouse embryos at E10 and E12 wereharvested using the PixCell II LCM System. Total RNA was extracted,amplified, labeled, and hybridized to the 430 2.0 mouse chip(Affymetrix) according to the manufacturer's instructions. Dataextracted from the images were analyzed using different softwareprograms. Regulated expression of selected genes was confirmed byreal-time PCR (RT-PCR).Results: Analysis of the microarray data from E10 and E12 lensesidentified 1,573 genes that showed a two fold or greater change inexpression level. Among these 1,573 genes, 956 genes were downregulatedand 617 were upregulated in E12 lenses. In addition to the upregulatedexpression of beta- and gamma-crystallin genes, genes that regulate thecell cycle showed significant changes of gene expression during the E10(lens pit) to E12 (primary fiber cell induction) time period. Genesinvolved in insulin-like growth factor (IGF) signaling and Wnt (a familyof secreted glycoproteins related to the Drosophila segment polaritygene, wingless, and to the proto-oncogene, int-1) signaling were alsodifferentially regulated. In particular, positive regulators of Wntsignaling were downregulated and negative regulators were upregulated,indicating that modulation of Wnt signaling is important for normal lensmorphogenesis.Conclusions: Our results provide new information about differentialregulation of gene expression during early lens development. Analysis ofglobal gene expression profiles in embryonic mouse lenses has allowed usto identify several molecular pathways that are differentially regulatedduring early lens development.

[学科分类] 生物化学/生物物理