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An RNA interference screen for genes required to shape the anteroposterior compartment boundary in Drosophila identifies the Eph receptor.

Umetsu D, Dunst S, Dahmann C - PLoS ONE (2014)

Bottom Line: Out of screening 3114 transgenic RNA interference lines targeting a total of 2863 genes, we identified a single novel candidate that interfered with the formation of a straight anteroposterior compartment boundary.Interestingly, the targeted gene encodes for the Eph receptor tyrosine kinase, an evolutionarily conserved family of signal transducers that has previously been shown to be important for maintaining straight compartment boundaries in vertebrate embryos.Our results identify a hitherto unknown role of the Eph receptor tyrosine kinase in Drosophila and suggest that Eph receptors have important functions in shaping compartment boundaries in both vertebrate and insect development.

View Article: PubMed Central - PubMed

Affiliation: Institute of Genetics, Technische Universität Dresden, Dresden, Germany.

ABSTRACT
The formation of straight compartment boundaries separating groups of cells with distinct fates and functions is an evolutionarily conserved strategy during animal development. The physical mechanisms that shape compartment boundaries have recently been further elucidated, however, the molecular mechanisms that underlie compartment boundary formation and maintenance remain poorly understood. Here, we report on the outcome of an RNA interference screen aimed at identifying novel genes involved in maintaining the straight shape of the anteroposterior compartment boundary in Drosophila wing imaginal discs. Out of screening 3114 transgenic RNA interference lines targeting a total of 2863 genes, we identified a single novel candidate that interfered with the formation of a straight anteroposterior compartment boundary. Interestingly, the targeted gene encodes for the Eph receptor tyrosine kinase, an evolutionarily conserved family of signal transducers that has previously been shown to be important for maintaining straight compartment boundaries in vertebrate embryos. Our results identify a hitherto unknown role of the Eph receptor tyrosine kinase in Drosophila and suggest that Eph receptors have important functions in shaping compartment boundaries in both vertebrate and insect development.

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Validation of assay.A–C. Wing discs expressing (A) no double-stranded RNA (control), (B) double-stranded RNA targeting smo, and (C) double-stranded RNA targeting ci in clones of cells marked by the expression of DsRed (red). Cells of the posterior compartment are labeled by expression of Venus under control of the engrailed gene (en-Venus, green). In (B) and (C), anterior clones along the AP boundary mis-segregate into the posterior territory of the wing disc (asterisks). Scale bar is 50 µm.
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pone-0114340-g002: Validation of assay.A–C. Wing discs expressing (A) no double-stranded RNA (control), (B) double-stranded RNA targeting smo, and (C) double-stranded RNA targeting ci in clones of cells marked by the expression of DsRed (red). Cells of the posterior compartment are labeled by expression of Venus under control of the engrailed gene (en-Venus, green). In (B) and (C), anterior clones along the AP boundary mis-segregate into the posterior territory of the wing disc (asterisks). Scale bar is 50 µm.

Mentions: To validate the assay we used RNAi lines targeting the genes smoothened and ci, two genes coding for a Hedgehog signal transducer and a transcription factor, respectively, that are known to be required in A cells to maintain the straight shape of the AP boundary [23], [24], [26]. Control clones of cells did not alter the shape of the AP boundary (Fig. 2A). By contrast, anterior clones of cells expressing double-stranded RNA targeting smo or ci altered the straight shape of the AP boundary and took up positions normally only occupied by posterior cells (Fig. 2B, C). This clonal behavior was reminiscent of clones of cells mutant for smo or ci in typical mosaic experiments using antibody stainings [23], [24], [26], indicating that these RNAi lines and the assay work faithfully.


An RNA interference screen for genes required to shape the anteroposterior compartment boundary in Drosophila identifies the Eph receptor.

Umetsu D, Dunst S, Dahmann C - PLoS ONE (2014)

Validation of assay.A–C. Wing discs expressing (A) no double-stranded RNA (control), (B) double-stranded RNA targeting smo, and (C) double-stranded RNA targeting ci in clones of cells marked by the expression of DsRed (red). Cells of the posterior compartment are labeled by expression of Venus under control of the engrailed gene (en-Venus, green). In (B) and (C), anterior clones along the AP boundary mis-segregate into the posterior territory of the wing disc (asterisks). Scale bar is 50 µm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4256218&req=5

pone-0114340-g002: Validation of assay.A–C. Wing discs expressing (A) no double-stranded RNA (control), (B) double-stranded RNA targeting smo, and (C) double-stranded RNA targeting ci in clones of cells marked by the expression of DsRed (red). Cells of the posterior compartment are labeled by expression of Venus under control of the engrailed gene (en-Venus, green). In (B) and (C), anterior clones along the AP boundary mis-segregate into the posterior territory of the wing disc (asterisks). Scale bar is 50 µm.
Mentions: To validate the assay we used RNAi lines targeting the genes smoothened and ci, two genes coding for a Hedgehog signal transducer and a transcription factor, respectively, that are known to be required in A cells to maintain the straight shape of the AP boundary [23], [24], [26]. Control clones of cells did not alter the shape of the AP boundary (Fig. 2A). By contrast, anterior clones of cells expressing double-stranded RNA targeting smo or ci altered the straight shape of the AP boundary and took up positions normally only occupied by posterior cells (Fig. 2B, C). This clonal behavior was reminiscent of clones of cells mutant for smo or ci in typical mosaic experiments using antibody stainings [23], [24], [26], indicating that these RNAi lines and the assay work faithfully.

Bottom Line: Out of screening 3114 transgenic RNA interference lines targeting a total of 2863 genes, we identified a single novel candidate that interfered with the formation of a straight anteroposterior compartment boundary.Interestingly, the targeted gene encodes for the Eph receptor tyrosine kinase, an evolutionarily conserved family of signal transducers that has previously been shown to be important for maintaining straight compartment boundaries in vertebrate embryos.Our results identify a hitherto unknown role of the Eph receptor tyrosine kinase in Drosophila and suggest that Eph receptors have important functions in shaping compartment boundaries in both vertebrate and insect development.

View Article: PubMed Central - PubMed

Affiliation: Institute of Genetics, Technische Universität Dresden, Dresden, Germany.

ABSTRACT
The formation of straight compartment boundaries separating groups of cells with distinct fates and functions is an evolutionarily conserved strategy during animal development. The physical mechanisms that shape compartment boundaries have recently been further elucidated, however, the molecular mechanisms that underlie compartment boundary formation and maintenance remain poorly understood. Here, we report on the outcome of an RNA interference screen aimed at identifying novel genes involved in maintaining the straight shape of the anteroposterior compartment boundary in Drosophila wing imaginal discs. Out of screening 3114 transgenic RNA interference lines targeting a total of 2863 genes, we identified a single novel candidate that interfered with the formation of a straight anteroposterior compartment boundary. Interestingly, the targeted gene encodes for the Eph receptor tyrosine kinase, an evolutionarily conserved family of signal transducers that has previously been shown to be important for maintaining straight compartment boundaries in vertebrate embryos. Our results identify a hitherto unknown role of the Eph receptor tyrosine kinase in Drosophila and suggest that Eph receptors have important functions in shaping compartment boundaries in both vertebrate and insect development.

Show MeSH