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The Drosophila CD2AP/CIN85 orthologue Cindr regulates junctions and cytoskeleton dynamics during tissue patterning.

Johnson RI, Seppa MJ, Cagan RL - J. Cell Biol. (2008)

Bottom Line: Reducing cindr activity leads to defects in local cell movement and, consequently, tissue patterning and cell death.Cindr activity is required for normal localization of Drosophila E-cadherin and Roughest, and we show additional physical and functional links to multiple components of the actin cytoskeleton, including the actin-capping proteins capping protein alpha and capping protein beta.Together, these data demonstrate that Cindr is involved in dynamic cell rearrangement in an emerging epithelium.

View Article: PubMed Central - PubMed

Affiliation: Department of Developmental and Regenerative Biology, Mount Sinai Medical School, New York, NY 10029, USA.

ABSTRACT
Developing tissues require cells to undergo intricate processes to shift into appropriate niches. This requires a functional connection between adhesion-mediating events at the cell surface and a cytoskeletal reorganization to permit directed movement. A small number of proteins are proposed to link these processes. Here, we identify one candidate, Cindr, the sole Drosophila melanogaster member of the CD2AP/CIN85 family (this family has been previously implicated in a variety of processes). Using D. melanogaster retina, we demonstrate that Cindr links cell surface junctions (E-cadherin) and adhesion (Roughest) with multiple components of the actin cytoskeleton. Reducing cindr activity leads to defects in local cell movement and, consequently, tissue patterning and cell death. Cindr activity is required for normal localization of Drosophila E-cadherin and Roughest, and we show additional physical and functional links to multiple components of the actin cytoskeleton, including the actin-capping proteins capping protein alpha and capping protein beta. Together, these data demonstrate that Cindr is involved in dynamic cell rearrangement in an emerging epithelium.

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cindr interacts with adhesion loci. (A) GMR>cindr-IR2 retina; DE-Cad was discontinuous (arrows). (B) GMR>cindr-IR2, de-cad. Mispatterning was rescued by expression of de-cad; cell membranes are elaborated and “frilly” (circled). (C) GMR>de-cad. Ectopic DE-Cad alone did not affect patterning. (D) shgR69/+/GMR>cindr-IR2. Enhanced mispatterning and discontinuous DE-Cad (arrows). (E–G) Adult eyes with black regions indicating tissue degeneration; genotypes are indicated. (H–J) Mild enhancement of cindr-IR2 in hbs or rst heterozygotes. (H) hbs459/GMR>cindr-IR2. (I) rst3/+;GMR>cindr-IR2. (J) rstCT/+;GMR>cindr-IR2. All tissue was dissected at 41 h APF; DE-Cad detection is shown. Adults were photographed 1 d after eclosion. Bars: (A–D and H–J) 10 μm; (E–G) 100 μm.
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fig4: cindr interacts with adhesion loci. (A) GMR>cindr-IR2 retina; DE-Cad was discontinuous (arrows). (B) GMR>cindr-IR2, de-cad. Mispatterning was rescued by expression of de-cad; cell membranes are elaborated and “frilly” (circled). (C) GMR>de-cad. Ectopic DE-Cad alone did not affect patterning. (D) shgR69/+/GMR>cindr-IR2. Enhanced mispatterning and discontinuous DE-Cad (arrows). (E–G) Adult eyes with black regions indicating tissue degeneration; genotypes are indicated. (H–J) Mild enhancement of cindr-IR2 in hbs or rst heterozygotes. (H) hbs459/GMR>cindr-IR2. (I) rst3/+;GMR>cindr-IR2. (J) rstCT/+;GMR>cindr-IR2. All tissue was dissected at 41 h APF; DE-Cad detection is shown. Adults were photographed 1 d after eclosion. Bars: (A–D and H–J) 10 μm; (E–G) 100 μm.

Mentions: Our results indicate a role for Cindr in mediating surface events. A subset of Cindr protein was found associated with DE-Cad (Figs. 1 E, S3, and S5), a central component of the AJ, and loss of cindr activity led to unstable junctions and abnormal cell movements. What is the relationship between Cindr and DE-Cad? Modifying a phenotype by removing one genomic copy of a gene is a standard and powerful technique to demonstrate that two loci function together in a common process. DE-Cad is encoded by the shotgun (shg) locus: the GMR>cindr-IR2 mispatterning phenotype (OMS = 7.2) was enhanced in a heterozygous shgR69/+ background (OMS = 10.8; Figs. 4 D and 2). In addition, large patches of black tissue were observed in the eyes of 68.3% (n = 97) of these adult flies (Fig. 4 G); this was a significant enhancement of the penetrance and extent of degenerative tissue that was observed in GMR>cindr-IR2 alone (12.6%, n = 254; Fig. 4 E). This blackened eye phenotype is suggestive of late epithelial cell death and indicates that the relationship between Cindr and DE-Cad extends to later stages of development.


The Drosophila CD2AP/CIN85 orthologue Cindr regulates junctions and cytoskeleton dynamics during tissue patterning.

Johnson RI, Seppa MJ, Cagan RL - J. Cell Biol. (2008)

cindr interacts with adhesion loci. (A) GMR>cindr-IR2 retina; DE-Cad was discontinuous (arrows). (B) GMR>cindr-IR2, de-cad. Mispatterning was rescued by expression of de-cad; cell membranes are elaborated and “frilly” (circled). (C) GMR>de-cad. Ectopic DE-Cad alone did not affect patterning. (D) shgR69/+/GMR>cindr-IR2. Enhanced mispatterning and discontinuous DE-Cad (arrows). (E–G) Adult eyes with black regions indicating tissue degeneration; genotypes are indicated. (H–J) Mild enhancement of cindr-IR2 in hbs or rst heterozygotes. (H) hbs459/GMR>cindr-IR2. (I) rst3/+;GMR>cindr-IR2. (J) rstCT/+;GMR>cindr-IR2. All tissue was dissected at 41 h APF; DE-Cad detection is shown. Adults were photographed 1 d after eclosion. Bars: (A–D and H–J) 10 μm; (E–G) 100 μm.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2290846&req=5

fig4: cindr interacts with adhesion loci. (A) GMR>cindr-IR2 retina; DE-Cad was discontinuous (arrows). (B) GMR>cindr-IR2, de-cad. Mispatterning was rescued by expression of de-cad; cell membranes are elaborated and “frilly” (circled). (C) GMR>de-cad. Ectopic DE-Cad alone did not affect patterning. (D) shgR69/+/GMR>cindr-IR2. Enhanced mispatterning and discontinuous DE-Cad (arrows). (E–G) Adult eyes with black regions indicating tissue degeneration; genotypes are indicated. (H–J) Mild enhancement of cindr-IR2 in hbs or rst heterozygotes. (H) hbs459/GMR>cindr-IR2. (I) rst3/+;GMR>cindr-IR2. (J) rstCT/+;GMR>cindr-IR2. All tissue was dissected at 41 h APF; DE-Cad detection is shown. Adults were photographed 1 d after eclosion. Bars: (A–D and H–J) 10 μm; (E–G) 100 μm.
Mentions: Our results indicate a role for Cindr in mediating surface events. A subset of Cindr protein was found associated with DE-Cad (Figs. 1 E, S3, and S5), a central component of the AJ, and loss of cindr activity led to unstable junctions and abnormal cell movements. What is the relationship between Cindr and DE-Cad? Modifying a phenotype by removing one genomic copy of a gene is a standard and powerful technique to demonstrate that two loci function together in a common process. DE-Cad is encoded by the shotgun (shg) locus: the GMR>cindr-IR2 mispatterning phenotype (OMS = 7.2) was enhanced in a heterozygous shgR69/+ background (OMS = 10.8; Figs. 4 D and 2). In addition, large patches of black tissue were observed in the eyes of 68.3% (n = 97) of these adult flies (Fig. 4 G); this was a significant enhancement of the penetrance and extent of degenerative tissue that was observed in GMR>cindr-IR2 alone (12.6%, n = 254; Fig. 4 E). This blackened eye phenotype is suggestive of late epithelial cell death and indicates that the relationship between Cindr and DE-Cad extends to later stages of development.

Bottom Line: Reducing cindr activity leads to defects in local cell movement and, consequently, tissue patterning and cell death.Cindr activity is required for normal localization of Drosophila E-cadherin and Roughest, and we show additional physical and functional links to multiple components of the actin cytoskeleton, including the actin-capping proteins capping protein alpha and capping protein beta.Together, these data demonstrate that Cindr is involved in dynamic cell rearrangement in an emerging epithelium.

View Article: PubMed Central - PubMed

Affiliation: Department of Developmental and Regenerative Biology, Mount Sinai Medical School, New York, NY 10029, USA.

ABSTRACT
Developing tissues require cells to undergo intricate processes to shift into appropriate niches. This requires a functional connection between adhesion-mediating events at the cell surface and a cytoskeletal reorganization to permit directed movement. A small number of proteins are proposed to link these processes. Here, we identify one candidate, Cindr, the sole Drosophila melanogaster member of the CD2AP/CIN85 family (this family has been previously implicated in a variety of processes). Using D. melanogaster retina, we demonstrate that Cindr links cell surface junctions (E-cadherin) and adhesion (Roughest) with multiple components of the actin cytoskeleton. Reducing cindr activity leads to defects in local cell movement and, consequently, tissue patterning and cell death. Cindr activity is required for normal localization of Drosophila E-cadherin and Roughest, and we show additional physical and functional links to multiple components of the actin cytoskeleton, including the actin-capping proteins capping protein alpha and capping protein beta. Together, these data demonstrate that Cindr is involved in dynamic cell rearrangement in an emerging epithelium.

Show MeSH
Related in: MedlinePlus