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Arginylation-dependent neural crest cell migration is essential for mouse development.

Kurosaka S, Leu NA, Zhang F, Bunte R, Saha S, Wang J, Guo C, He W, Kashina A - PLoS Genet. (2010)

Bottom Line: Recent studies of protein arginylation implicated this poorly understood posttranslational modification in the functioning of actin cytoskeleton and in cell migration in culture.Wnt1-Ate1 pups have prominent defects, including short palate and altered opening to the nasopharynx, and cranial defects that likely contribute to the abnormal breathing and early death.Analysis of neural crest cell movement patterns in situ and cell motility in culture shows an overall delay in the migration of Ate1 knockout cells that is likely regulated by intracellular mechanisms rather than extracellular signaling events.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.

ABSTRACT
Coordinated cell migration during development is crucial for morphogenesis and largely relies on cells of the neural crest lineage that migrate over long distances to give rise to organs and tissues throughout the body. Recent studies of protein arginylation implicated this poorly understood posttranslational modification in the functioning of actin cytoskeleton and in cell migration in culture. Knockout of arginyltransferase (Ate1) in mice leads to embryonic lethality and severe heart defects that are reminiscent of cell migration-dependent phenotypes seen in other mouse models. To test the hypothesis that arginylation regulates cell migration during morphogenesis, we produced Wnt1-Cre Ate1 conditional knockout mice (Wnt1-Ate1), with Ate1 deletion in the neural crest cells driven by Wnt1 promoter. Wnt1-Ate1 mice die at birth and in the first 2-3 weeks after birth with severe breathing problems and with growth and behavioral retardation. Wnt1-Ate1 pups have prominent defects, including short palate and altered opening to the nasopharynx, and cranial defects that likely contribute to the abnormal breathing and early death. Analysis of neural crest cell movement patterns in situ and cell motility in culture shows an overall delay in the migration of Ate1 knockout cells that is likely regulated by intracellular mechanisms rather than extracellular signaling events. Taken together, our data suggest that arginylation plays a general role in the migration of the neural crest cells in development by regulating the molecular machinery that underlies cell migration through tissues and organs during morphogenesis.

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Related in: MedlinePlus

Wnt1-Ate1 mice have defects in the frontal bones.(A) Top view of the skulls of the control (WT, top) and Wnt1-Ate1 (KO, bottom) pups at P0 stained with alizarin red S and alcian blue 8GS show a significant gap on top of the skull in the mutant pups, due to the significantly reduced frontal bones. Left images show the same pictures as those shown on right with the edges of the gap outlined. Arrowheads indicate the point of the opening of the gap used for the measurements shown in (B). (B) Measurement of the ratio of the gap width to skull width along the same axis (gap/skull) at P0 show that in the mutant (KO) unlike the control (WT) the gap occupies on average more than 1/3 of the skull (ratio 0.334+/−0.018 (SEM, n = 5) in the mutant vs. 0.076+/−0.016 (SEM, n = 12) in wild-type). (C) Skulls of adult Wnt1-Ate1 mice compared to their littermate controls show frontal bone abnormalities, including abnormal shape, incomplete cranial suture (arrows), and abnormal suture between the frontal bones (arrowheads). 2 wild-type and 2 mutant animals were analyzed.
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pgen-1000878-g003: Wnt1-Ate1 mice have defects in the frontal bones.(A) Top view of the skulls of the control (WT, top) and Wnt1-Ate1 (KO, bottom) pups at P0 stained with alizarin red S and alcian blue 8GS show a significant gap on top of the skull in the mutant pups, due to the significantly reduced frontal bones. Left images show the same pictures as those shown on right with the edges of the gap outlined. Arrowheads indicate the point of the opening of the gap used for the measurements shown in (B). (B) Measurement of the ratio of the gap width to skull width along the same axis (gap/skull) at P0 show that in the mutant (KO) unlike the control (WT) the gap occupies on average more than 1/3 of the skull (ratio 0.334+/−0.018 (SEM, n = 5) in the mutant vs. 0.076+/−0.016 (SEM, n = 12) in wild-type). (C) Skulls of adult Wnt1-Ate1 mice compared to their littermate controls show frontal bone abnormalities, including abnormal shape, incomplete cranial suture (arrows), and abnormal suture between the frontal bones (arrowheads). 2 wild-type and 2 mutant animals were analyzed.

Mentions: Since neural crest cells contribute to the formation of bone and cartilage in the head, we next examined the skeletons of newborn Wnt1-Ate1 and control mice at P0 by staining with alizarin red S and alcian blue 8GS that interact with bones and cartilage to color them red and blue, respectively [41] (Figure 3). While the overall bone structure and skeletal architecture appeared normal in Wnt1-Ate1 mice (data not shown), prominent abnormalities were seen in the development of the frontal bones, the neural-crest-derived parts of craniofacial skeleton that contribute to the top of the skull [32],[42]. In control mice, frontal bones came close together, leaving only a narrow slit along the top of the skull (Figure 3A, top images). In contrast, in the mutant mice, frontal bones appeared smaller and narrower and were unable to meet on top of the skull, leaving a wide gap that exposed the cranial cavity beneath (Figure 3A, bottom images). Measurements of the ratios between the width of the gap and the width of the skull showed that in the mutant the gap occupied on average almost 1/3 of the skull width, an area almost 4 times larger than in wild-type (Figure 3B). This defect was observed in all the analyzed mutants, even those that did not exhibit breathing defects at birth.


Arginylation-dependent neural crest cell migration is essential for mouse development.

Kurosaka S, Leu NA, Zhang F, Bunte R, Saha S, Wang J, Guo C, He W, Kashina A - PLoS Genet. (2010)

Wnt1-Ate1 mice have defects in the frontal bones.(A) Top view of the skulls of the control (WT, top) and Wnt1-Ate1 (KO, bottom) pups at P0 stained with alizarin red S and alcian blue 8GS show a significant gap on top of the skull in the mutant pups, due to the significantly reduced frontal bones. Left images show the same pictures as those shown on right with the edges of the gap outlined. Arrowheads indicate the point of the opening of the gap used for the measurements shown in (B). (B) Measurement of the ratio of the gap width to skull width along the same axis (gap/skull) at P0 show that in the mutant (KO) unlike the control (WT) the gap occupies on average more than 1/3 of the skull (ratio 0.334+/−0.018 (SEM, n = 5) in the mutant vs. 0.076+/−0.016 (SEM, n = 12) in wild-type). (C) Skulls of adult Wnt1-Ate1 mice compared to their littermate controls show frontal bone abnormalities, including abnormal shape, incomplete cranial suture (arrows), and abnormal suture between the frontal bones (arrowheads). 2 wild-type and 2 mutant animals were analyzed.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC2837401&req=5

pgen-1000878-g003: Wnt1-Ate1 mice have defects in the frontal bones.(A) Top view of the skulls of the control (WT, top) and Wnt1-Ate1 (KO, bottom) pups at P0 stained with alizarin red S and alcian blue 8GS show a significant gap on top of the skull in the mutant pups, due to the significantly reduced frontal bones. Left images show the same pictures as those shown on right with the edges of the gap outlined. Arrowheads indicate the point of the opening of the gap used for the measurements shown in (B). (B) Measurement of the ratio of the gap width to skull width along the same axis (gap/skull) at P0 show that in the mutant (KO) unlike the control (WT) the gap occupies on average more than 1/3 of the skull (ratio 0.334+/−0.018 (SEM, n = 5) in the mutant vs. 0.076+/−0.016 (SEM, n = 12) in wild-type). (C) Skulls of adult Wnt1-Ate1 mice compared to their littermate controls show frontal bone abnormalities, including abnormal shape, incomplete cranial suture (arrows), and abnormal suture between the frontal bones (arrowheads). 2 wild-type and 2 mutant animals were analyzed.
Mentions: Since neural crest cells contribute to the formation of bone and cartilage in the head, we next examined the skeletons of newborn Wnt1-Ate1 and control mice at P0 by staining with alizarin red S and alcian blue 8GS that interact with bones and cartilage to color them red and blue, respectively [41] (Figure 3). While the overall bone structure and skeletal architecture appeared normal in Wnt1-Ate1 mice (data not shown), prominent abnormalities were seen in the development of the frontal bones, the neural-crest-derived parts of craniofacial skeleton that contribute to the top of the skull [32],[42]. In control mice, frontal bones came close together, leaving only a narrow slit along the top of the skull (Figure 3A, top images). In contrast, in the mutant mice, frontal bones appeared smaller and narrower and were unable to meet on top of the skull, leaving a wide gap that exposed the cranial cavity beneath (Figure 3A, bottom images). Measurements of the ratios between the width of the gap and the width of the skull showed that in the mutant the gap occupied on average almost 1/3 of the skull width, an area almost 4 times larger than in wild-type (Figure 3B). This defect was observed in all the analyzed mutants, even those that did not exhibit breathing defects at birth.

Bottom Line: Recent studies of protein arginylation implicated this poorly understood posttranslational modification in the functioning of actin cytoskeleton and in cell migration in culture.Wnt1-Ate1 pups have prominent defects, including short palate and altered opening to the nasopharynx, and cranial defects that likely contribute to the abnormal breathing and early death.Analysis of neural crest cell movement patterns in situ and cell motility in culture shows an overall delay in the migration of Ate1 knockout cells that is likely regulated by intracellular mechanisms rather than extracellular signaling events.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.

ABSTRACT
Coordinated cell migration during development is crucial for morphogenesis and largely relies on cells of the neural crest lineage that migrate over long distances to give rise to organs and tissues throughout the body. Recent studies of protein arginylation implicated this poorly understood posttranslational modification in the functioning of actin cytoskeleton and in cell migration in culture. Knockout of arginyltransferase (Ate1) in mice leads to embryonic lethality and severe heart defects that are reminiscent of cell migration-dependent phenotypes seen in other mouse models. To test the hypothesis that arginylation regulates cell migration during morphogenesis, we produced Wnt1-Cre Ate1 conditional knockout mice (Wnt1-Ate1), with Ate1 deletion in the neural crest cells driven by Wnt1 promoter. Wnt1-Ate1 mice die at birth and in the first 2-3 weeks after birth with severe breathing problems and with growth and behavioral retardation. Wnt1-Ate1 pups have prominent defects, including short palate and altered opening to the nasopharynx, and cranial defects that likely contribute to the abnormal breathing and early death. Analysis of neural crest cell movement patterns in situ and cell motility in culture shows an overall delay in the migration of Ate1 knockout cells that is likely regulated by intracellular mechanisms rather than extracellular signaling events. Taken together, our data suggest that arginylation plays a general role in the migration of the neural crest cells in development by regulating the molecular machinery that underlies cell migration through tissues and organs during morphogenesis.

Show MeSH
Related in: MedlinePlus