TIE-DYE: a combinatorial marking system to visualize and genetically manipulate clones during development in Drosophila melanogaster

Abstract

SUMMARYTwo types of information are particularly valuable in understanding the development of a tissue or an organ from a small population of founder cells. First, it is useful to know the composition of the final structure in terms the contribution of individual founder cells. Second, it is important to understand cell-cell interactions. To facilitate the study of both of these aspects of organ development at a tissue-wide level, we have developed a method, TIE-DYE, that allows simultaneous lineage tracing of multiple cell populations as well as the genetic manipulation of a subset of these populations. Seven uniquely marked categories of cells are produced by sitedirected recombination of three independent cassettes. We have used the TIE-DYE method to estimate the number of founder cells that give rise to the wing-imaginal disc during normal development and following compensatory growth caused by X-ray irradiation of the founder cells. We also show that four out of the seven types of marked clones can be genetically manipulated by gene overexpression or RNAi knockdown, allowing an assessment of the consequences of these manipulations on the entire wing disc. We demonstrate the utility of this system in studying the consequences of alterations in growth, patterning and cell-cell affinity.

Modulation of junction tension by tumor-suppressors and proto-oncogenes regulates cell-cell contacts

“…Numerous tumor suppressors and proto-oncogenes share the common feature of promoting cell segregation apparent by the rounding of somatic clones upon their loss or gain of functions, respectively (Adler et al, 1998;Johnston et al, 1999;Edgar, 2000, 2002;Garoia et al, 2000;Baena-Lopez et al, 2005;Mao et al, 2006Mao et al, , 2011Worley et al, 2013). Here, we show that loss of Ft/Ds and Hippo tumor-suppressor pathways as well as gain of function of the protooncogenes Myc, Ras and Yki lead to changes in cell junction tension.…”

“…The analyses of these functions have led to important advances in our understanding of tissue development and homeostasis as well as pathologies, including tumorigenesis (for reviews, see Zhao et al, 2011;Patel and Edgar, 2014;Baillon and Basler, 2014). In Drosophila, analysis of the sizes and shapes of somatic clones affecting tumor suppressor and proto-oncogene activities is instrumental to understanding their contribution in tissue morphogenesis, organization and homeostasis (Resino et al, 2002;Baena-Lopez et al, 2005;Mao et al, 2011;Wartlick et al, 2011;Kuchen et al, 2012;Worley et al, 2013;Restrepo et al, 2014;Heemskerk et al, 2014). Accordingly, somatic mutant clones are essential for unveiling how tumor suppressor and proto-oncogene activities modulate tissue proliferation, growth, cell-cell interactions and cell competition (for a review, see Wagstaff et al, 2013).…”

“…Accordingly, somatic mutant clones are essential for unveiling how tumor suppressor and proto-oncogene activities modulate tissue proliferation, growth, cell-cell interactions and cell competition (for a review, see Wagstaff et al, 2013). In particular, the functions of tumor suppressors and proto-oncogenes in tissue organization and morphogenesis have often been recognized as their respective loss and gain of function leads to the formation of a rounded group of mutant cells (somatic clones) having a smooth boundary with the surrounding wild-type (wt) cells and thus reducing their contacts with neighboring wt tissue (Justice et al, 1995;Edgar, 2000, 2002;Baena-Lopez et al, 2005;Mao et al, 2006;Worley et al, 2013). This property is shared by the Ras and Myc proto-oncogenes as well as components of the Fat/Dachsous (Ft/Ds) and Hippo pathways (Justice et al, 1995;Adler et al, 1998;Johnston et al, 1999;Edgar, 2000, 2002;Garoia et al, 2000;Baena-Lopez et al, 2005;Mao et al, 2011;Worley et al, 2013).…”

Dynamic clonal analysis based on chronic in vivo imaging allows multiscale quantification of growth in the Drosophila wing disc

“…In combination with genetic mutations, it also facilitated dissection of the pathways that control size and shape. New techniques recently expanded the toolbox of clonal analysis to multicolor lineages inspired by the Brainbow strategy (Livet et al, 2007;Boulina et al, 2013;Worley et al, 2013). The established method for inferring growth from clones is to dissect the discs out of the larvae, fix and then image them.…”

Raeppli: a whole-tissue labeling tool for live imaging of Drosophila development

“…The primordium of the Drosophila melanogaster wing is a group of ~25-50 cells set aside during embryogenesis. During the following 5 days of larval development, the primordium grows to form a sac-like structure of ~31,000 cells, called the wing imaginal disc (Madhavan and Schneiderman, 1977;Martín et al, 2009;Worley et al, 2013). During metamorphosis, most of the larval cells undergo histolysis and the wing imaginal discs evert to form the adult wing and notum structures (Cohen et al, 1993;Merriam, 1978;Milán et al, 1996).…”