title: Image analysis and empirical modeling of gene and protein expression.
abstract: 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.
authors.affiliation: Division of Biology, California Institute of Technology, Pasadena, CA 91125, USA.
authors.forename: Nathanie
authors.initials: N
authors.lastname: Trisnadi
authors.forename: Alphan
authors.initials: A
authors.lastname: Altinok
authors.forename: Angelike
authors.initials: A
authors.lastname: Stathopoulos
authors.forename: Gregory T
authors.initials: GT
authors.lastname: Reeves
mesh_terms: D000818:Animals; D019521:Body Patterning / Q000235:genetics; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism; D004331:Drosophila melanogaster / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D018507:Gene Expression Regulation, Developmental; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D007091:Image Processing, Computer-Assisted / Q000706:statistics & numerical data*; D017403:In Situ Hybridization; D018613:Microscopy, Confocal; D008957:Models, Genetic; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins; D018398:Homeodomain Proteins; D009687:Nuclear Proteins; D010750:Phosphoproteins; D014157:Transcription Factors; C062236:dl protein, Drosophila; C094797:vnd protein, Drosophila
keywords: DV; Dorsal–ventral patterning; Drosophila melanogaster; Empirical modeling; GFP; Gene expression; Protein gradient; Quantitative measurements; YFP; dl; dorsal; dorsal–ventral; green fluorescent protein; nc; nuclear cycles; short-gastrulation; sog; ventral-neuroblasts defective; vnd; yellow fluorescent protein; zen; zerknüllt