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Dishevelled-1 regulates microtubule stability: a new function mediated by glycogen synthase kinase-3beta.

Krylova O, Messenger MJ, Salinas PC - J. Cell Biol. (2000)

Bottom Line: The microtubule stabilizing function of DVL-1 is mimicked by lithium-mediated inhibition of glycogen synthase kinase-3beta (GSK-3beta) and blocked by expression of GSK-3beta.These findings suggest that DVL-1, through GSK-3beta, can regulate microtubule dynamics.This new function of DVL-1 in controlling microtubule stability may have important implications for Dishevelled proteins in regulating cell polarity.

View Article: PubMed Central - PubMed

Affiliation: The Randall Institute, King's College London, London, United Kingdom.

ABSTRACT
Dishevelled has been implicated in the regulation of cell fate decisions, cell polarity, and neuronal function. However, the mechanism of Dishevelled action remains poorly understood. Here we examine the cellular localization and function of the mouse Dishevelled protein, DVL-1. Endogenous DVL-1 colocalizes with axonal microtubules and sediments with brain microtubules. Expression of DVL-1 protects stable microtubules from depolymerization by nocodazole in both dividing cells and differentiated neuroblastoma cells. Deletion analyses reveal that the PDZ domain, but not the DEP domain, of DVL-1 is required for microtubule stabilization. The microtubule stabilizing function of DVL-1 is mimicked by lithium-mediated inhibition of glycogen synthase kinase-3beta (GSK-3beta) and blocked by expression of GSK-3beta. These findings suggest that DVL-1, through GSK-3beta, can regulate microtubule dynamics. This new function of DVL-1 in controlling microtubule stability may have important implications for Dishevelled proteins in regulating cell polarity.

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DVL-1 expression in postnatal mouse brain. Brain sections were immunostained for DVL-1 (A–J). The highest level of DVL-1 was detected in neurons of the cerebral cortex, hippocampus, pons, and cerebellum (A–C and I). (A, inset) DVL-1 localization in cell bodies and processes of pyramidal neurons. In P0 cerebellum, DVL-1 is expressed in the EGL and forming IGL (D). At P7, DVL-1 immunoreactivity increases in the EGL (E). At P14, DVL-1 was mainly localized in the granule cell layer (GCL) and in the Purkinje cell (PC) layer (F). This pattern of DVL-1 expression was maintained throughout life (G–I). A low level of DVL-1 expression was detected in the molecular layer (ML) from P21 (G–I). At P21, DVL-1 is mainly localized in granule cells and Purkinje cells cell bodies, as shown at higher magnification (J). No immunoreactivity was detected in the cerebellum of adult Dvl-1  mutant mice (K). Preincubation of the antibody with DVL-46 peptide completely abolished immunostaining in adult cerebellum (L). AD, adult; DG, dentate gyrus. Bar, 100 μM.
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Figure 1: DVL-1 expression in postnatal mouse brain. Brain sections were immunostained for DVL-1 (A–J). The highest level of DVL-1 was detected in neurons of the cerebral cortex, hippocampus, pons, and cerebellum (A–C and I). (A, inset) DVL-1 localization in cell bodies and processes of pyramidal neurons. In P0 cerebellum, DVL-1 is expressed in the EGL and forming IGL (D). At P7, DVL-1 immunoreactivity increases in the EGL (E). At P14, DVL-1 was mainly localized in the granule cell layer (GCL) and in the Purkinje cell (PC) layer (F). This pattern of DVL-1 expression was maintained throughout life (G–I). A low level of DVL-1 expression was detected in the molecular layer (ML) from P21 (G–I). At P21, DVL-1 is mainly localized in granule cells and Purkinje cells cell bodies, as shown at higher magnification (J). No immunoreactivity was detected in the cerebellum of adult Dvl-1 mutant mice (K). Preincubation of the antibody with DVL-46 peptide completely abolished immunostaining in adult cerebellum (L). AD, adult; DG, dentate gyrus. Bar, 100 μM.

Mentions: To examine the function of DVL-1 in neuronal development, we generated an affinity-purified polyclonal antibody against a peptide encoding the 46 carboxy-terminal amino acids of DVL-1. Using this antibody, we found that DVL-1 is localized to several neuronal populations of the adult mouse CNS. High levels of DVL-1 protein were found in neurons of the cerebral cortex, hippocampus, pons, and cerebellum (Fig. 1, A–J). In the cortex, DVL-1 is localized to the cell body and processes of neurons (Fig. 1 A). High levels of DVL-1 were found in pyramidal neurons (Fig. 1 A, insert). In the cerebellum, we examined in more detail the distribution of DVL-1 during postnatal cerebellar development (Fig. 1, D–J). At birth, DVL-1 was found in the external granular cell layer (EGL) and in the forming internal granule cell layer (IGL) of the cerebellum (Fig. 1 D). At P7, the level of DVL-1 is higher in the EGL as granule cells migrate to the IGL. DVL-1 was also observed in the forming IGL (Fig. 1 E). By P14, DVL-1 is highly expressed in the granule cell layer and this expression is maintained in adult life (Fig. 1, F–J). DVL-1 is also expressed in the Purkinje cell layer, when Purkinje cells begin to mature and develop a complex dendritic tree (Fig. 1, F–J). From P21, low levels of DVL-1 were found in the molecular layer (Fig. 1, G–I). In Dvl-1 mutant mice, no immunoreactivity was observed (Fig. 1 K). Preincubation of the antibody with the DVL-46 peptide completely abolishes DVL-1 staining in adult mouse cerebellum (Fig. 1 L). These results show that DVL-1 is mainly localized to neurons throughout life.


Dishevelled-1 regulates microtubule stability: a new function mediated by glycogen synthase kinase-3beta.

Krylova O, Messenger MJ, Salinas PC - J. Cell Biol. (2000)

DVL-1 expression in postnatal mouse brain. Brain sections were immunostained for DVL-1 (A–J). The highest level of DVL-1 was detected in neurons of the cerebral cortex, hippocampus, pons, and cerebellum (A–C and I). (A, inset) DVL-1 localization in cell bodies and processes of pyramidal neurons. In P0 cerebellum, DVL-1 is expressed in the EGL and forming IGL (D). At P7, DVL-1 immunoreactivity increases in the EGL (E). At P14, DVL-1 was mainly localized in the granule cell layer (GCL) and in the Purkinje cell (PC) layer (F). This pattern of DVL-1 expression was maintained throughout life (G–I). A low level of DVL-1 expression was detected in the molecular layer (ML) from P21 (G–I). At P21, DVL-1 is mainly localized in granule cells and Purkinje cells cell bodies, as shown at higher magnification (J). No immunoreactivity was detected in the cerebellum of adult Dvl-1  mutant mice (K). Preincubation of the antibody with DVL-46 peptide completely abolished immunostaining in adult cerebellum (L). AD, adult; DG, dentate gyrus. Bar, 100 μM.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2189803&req=5

Figure 1: DVL-1 expression in postnatal mouse brain. Brain sections were immunostained for DVL-1 (A–J). The highest level of DVL-1 was detected in neurons of the cerebral cortex, hippocampus, pons, and cerebellum (A–C and I). (A, inset) DVL-1 localization in cell bodies and processes of pyramidal neurons. In P0 cerebellum, DVL-1 is expressed in the EGL and forming IGL (D). At P7, DVL-1 immunoreactivity increases in the EGL (E). At P14, DVL-1 was mainly localized in the granule cell layer (GCL) and in the Purkinje cell (PC) layer (F). This pattern of DVL-1 expression was maintained throughout life (G–I). A low level of DVL-1 expression was detected in the molecular layer (ML) from P21 (G–I). At P21, DVL-1 is mainly localized in granule cells and Purkinje cells cell bodies, as shown at higher magnification (J). No immunoreactivity was detected in the cerebellum of adult Dvl-1 mutant mice (K). Preincubation of the antibody with DVL-46 peptide completely abolished immunostaining in adult cerebellum (L). AD, adult; DG, dentate gyrus. Bar, 100 μM.
Mentions: To examine the function of DVL-1 in neuronal development, we generated an affinity-purified polyclonal antibody against a peptide encoding the 46 carboxy-terminal amino acids of DVL-1. Using this antibody, we found that DVL-1 is localized to several neuronal populations of the adult mouse CNS. High levels of DVL-1 protein were found in neurons of the cerebral cortex, hippocampus, pons, and cerebellum (Fig. 1, A–J). In the cortex, DVL-1 is localized to the cell body and processes of neurons (Fig. 1 A). High levels of DVL-1 were found in pyramidal neurons (Fig. 1 A, insert). In the cerebellum, we examined in more detail the distribution of DVL-1 during postnatal cerebellar development (Fig. 1, D–J). At birth, DVL-1 was found in the external granular cell layer (EGL) and in the forming internal granule cell layer (IGL) of the cerebellum (Fig. 1 D). At P7, the level of DVL-1 is higher in the EGL as granule cells migrate to the IGL. DVL-1 was also observed in the forming IGL (Fig. 1 E). By P14, DVL-1 is highly expressed in the granule cell layer and this expression is maintained in adult life (Fig. 1, F–J). DVL-1 is also expressed in the Purkinje cell layer, when Purkinje cells begin to mature and develop a complex dendritic tree (Fig. 1, F–J). From P21, low levels of DVL-1 were found in the molecular layer (Fig. 1, G–I). In Dvl-1 mutant mice, no immunoreactivity was observed (Fig. 1 K). Preincubation of the antibody with the DVL-46 peptide completely abolishes DVL-1 staining in adult mouse cerebellum (Fig. 1 L). These results show that DVL-1 is mainly localized to neurons throughout life.

Bottom Line: The microtubule stabilizing function of DVL-1 is mimicked by lithium-mediated inhibition of glycogen synthase kinase-3beta (GSK-3beta) and blocked by expression of GSK-3beta.These findings suggest that DVL-1, through GSK-3beta, can regulate microtubule dynamics.This new function of DVL-1 in controlling microtubule stability may have important implications for Dishevelled proteins in regulating cell polarity.

View Article: PubMed Central - PubMed

Affiliation: The Randall Institute, King's College London, London, United Kingdom.

ABSTRACT
Dishevelled has been implicated in the regulation of cell fate decisions, cell polarity, and neuronal function. However, the mechanism of Dishevelled action remains poorly understood. Here we examine the cellular localization and function of the mouse Dishevelled protein, DVL-1. Endogenous DVL-1 colocalizes with axonal microtubules and sediments with brain microtubules. Expression of DVL-1 protects stable microtubules from depolymerization by nocodazole in both dividing cells and differentiated neuroblastoma cells. Deletion analyses reveal that the PDZ domain, but not the DEP domain, of DVL-1 is required for microtubule stabilization. The microtubule stabilizing function of DVL-1 is mimicked by lithium-mediated inhibition of glycogen synthase kinase-3beta (GSK-3beta) and blocked by expression of GSK-3beta. These findings suggest that DVL-1, through GSK-3beta, can regulate microtubule dynamics. This new function of DVL-1 in controlling microtubule stability may have important implications for Dishevelled proteins in regulating cell polarity.

Show MeSH
Related in: MedlinePlus