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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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Clones of cells expressing double-stranded RNA targeting the Eph gene distort the AP boundary.A–C. Wing imaginal discs displaying clones of cells expressing double-stranded RNA targeting (A–B) Eph or (C) FasIII. The clones of cells are identified by co-expression of DsRed (red); cells of the posterior compartment are labeled by expression of Venus under control of the engrailed gene (en-Venus, green). (A) Individual clones abutting the AP boundary from either the A or P compartment do not influence the shape of the boundary (asterisk). (B) Two clones expressing EphdsRNA (ID-Number 4771 (VDRC)) located in different compartments sharing a common interface along the AP boundary locally distort the shape of the AP boundary (asterisks). (C) Two clones expressing FasIIIdsRNA located in different compartments sharing a common interface along the AP boundary do not distort the shape of the AP boundary (asterisks). Scale bar is 50 µm.
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pone-0114340-g005: Clones of cells expressing double-stranded RNA targeting the Eph gene distort the AP boundary.A–C. Wing imaginal discs displaying clones of cells expressing double-stranded RNA targeting (A–B) Eph or (C) FasIII. The clones of cells are identified by co-expression of DsRed (red); cells of the posterior compartment are labeled by expression of Venus under control of the engrailed gene (en-Venus, green). (A) Individual clones abutting the AP boundary from either the A or P compartment do not influence the shape of the boundary (asterisk). (B) Two clones expressing EphdsRNA (ID-Number 4771 (VDRC)) located in different compartments sharing a common interface along the AP boundary locally distort the shape of the AP boundary (asterisks). (C) Two clones expressing FasIIIdsRNA located in different compartments sharing a common interface along the AP boundary do not distort the shape of the AP boundary (asterisks). Scale bar is 50 µm.

Mentions: The roundish shape of clones is a signature of cell sorting [44]. We therefore re-screened RNAi lines that in the primary screen displayed a rounded shape for causing defects in the shape of the A/P boundary. In no case did we find that individual clones of cells located along the AP boundary influenced boundary shape (e.g. Fig. 5A, and Table S1). Interestingly, however, we found that expression of three RNAi lines, 43454, 10064, and 6545, targeting the CG10176, CG9416 and Eph genes, respectively, resulted in the following phenotype: when two clones located in different compartments shared a common interface along the AP boundary, then the shape of the AP boundary was locally distorted (Fig. 5B and data not shown). Abutting clones of cells from different compartments displaying smooth clone borders as a result of expression of double-stranded RNA targeting PlexA (n = 10 clones), ds (n = 21 clones), or FasIII (n = 6 clones) did not influence the shape of the AP boundary (Fig. 5C, data not shown). This suggests that two abutting clones with smooth clone borders do not per se disturb the shape of the AP boundary.


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)

Clones of cells expressing double-stranded RNA targeting the Eph gene distort the AP boundary.A–C. Wing imaginal discs displaying clones of cells expressing double-stranded RNA targeting (A–B) Eph or (C) FasIII. The clones of cells are identified by co-expression of DsRed (red); cells of the posterior compartment are labeled by expression of Venus under control of the engrailed gene (en-Venus, green). (A) Individual clones abutting the AP boundary from either the A or P compartment do not influence the shape of the boundary (asterisk). (B) Two clones expressing EphdsRNA (ID-Number 4771 (VDRC)) located in different compartments sharing a common interface along the AP boundary locally distort the shape of the AP boundary (asterisks). (C) Two clones expressing FasIIIdsRNA located in different compartments sharing a common interface along the AP boundary do not distort the shape of the AP boundary (asterisks). Scale bar is 50 µm.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0114340-g005: Clones of cells expressing double-stranded RNA targeting the Eph gene distort the AP boundary.A–C. Wing imaginal discs displaying clones of cells expressing double-stranded RNA targeting (A–B) Eph or (C) FasIII. The clones of cells are identified by co-expression of DsRed (red); cells of the posterior compartment are labeled by expression of Venus under control of the engrailed gene (en-Venus, green). (A) Individual clones abutting the AP boundary from either the A or P compartment do not influence the shape of the boundary (asterisk). (B) Two clones expressing EphdsRNA (ID-Number 4771 (VDRC)) located in different compartments sharing a common interface along the AP boundary locally distort the shape of the AP boundary (asterisks). (C) Two clones expressing FasIIIdsRNA located in different compartments sharing a common interface along the AP boundary do not distort the shape of the AP boundary (asterisks). Scale bar is 50 µm.
Mentions: The roundish shape of clones is a signature of cell sorting [44]. We therefore re-screened RNAi lines that in the primary screen displayed a rounded shape for causing defects in the shape of the A/P boundary. In no case did we find that individual clones of cells located along the AP boundary influenced boundary shape (e.g. Fig. 5A, and Table S1). Interestingly, however, we found that expression of three RNAi lines, 43454, 10064, and 6545, targeting the CG10176, CG9416 and Eph genes, respectively, resulted in the following phenotype: when two clones located in different compartments shared a common interface along the AP boundary, then the shape of the AP boundary was locally distorted (Fig. 5B and data not shown). Abutting clones of cells from different compartments displaying smooth clone borders as a result of expression of double-stranded RNA targeting PlexA (n = 10 clones), ds (n = 21 clones), or FasIII (n = 6 clones) did not influence the shape of the AP boundary (Fig. 5C, data not shown). This suggests that two abutting clones with smooth clone borders do not per se disturb the shape of the AP boundary.

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