کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1993620 | 1064689 | 2013 | 11 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Image analysis and empirical modeling of gene and protein expression Image analysis and empirical modeling of gene and protein expression](/preview/png/1993620.png)
Protein gradients and gene expression patterns are major determinants in the differentiation and fate map of the developing embryo. Here we discuss computational methods to quantitatively measure the positions of gene expression domains and the gradients of protein expression along the dorsal–ventral axis in the Drosophila embryo. Our methodology involves three layers of data. The first layer, or the primary data, consists of z-stack confocal images of embryos processed by in situ hybridization and/or antibody stainings. The secondary data are relationships between location, usually an x-axis coordinate, and fluorescent intensity of gene or protein detection. Tertiary data comprise the optimal parameters that arise from fits of the secondary data to empirical models. The tertiary data are useful to distill large datasets of imaged embryos down to a tractable number of conceptually useful parameters. This analysis allows us to detect subtle phenotypes and is adaptable to any set of genes or proteins with a canonical pattern. For example, we show how insights into the Dorsal transcription factor protein gradient and its target gene ventral-neuroblasts defective (vnd) were obtained using such quantitative approaches.
► Morphogen gradients guide gene expression in the developing Drosophila embryo.
► We develop computational methods to characterize domain boundaries.
► This method yields parameters that can be compared in large datasets.
► Canonical curve fitting produces high-confidence measurements resistant to noise.
► Our quantitative analysis can detect subtle phenotypes in mutant embryos.
Journal: Methods - Volume 62, Issue 1, 15 July 2013, Pages 68–78