Arber, S. et al. Regulation of actin dynamics through phosphorylation of cofilin by LIM-kinase. Nature 393, 805-809
ABSTRACT Cell division, cell motility and the formation and maintenance of specialized structures in differentiated cells depend directly on the regulated dynamics of the actin cytoskeleton. To understand the mechanisms of these basic cellular processes, the signalling pathways that link external signals to the regulation of the actin cytoskeleton need to be characterized. Here we identify a pathway for the regulation of cofilin, a ubiquitous actin-binding protein that is essential for effective depolymerization of actin filaments. LIM-kinase 1, also known as KIZ, is a protein kinase with two amino-terminal LIM motifs that induces stabilization of F-actin structures in transfected cells. Dominant-negative LIM-kinasel inhibits the accumulation of the F-actin. Phosphorylation experiments in vivo and in vitro provide evidence that cofilin is a physiological substrate of LIM-kinase 1. Phosphorylation by LIM-kinase 1 inactivates cofilin, leading to accumulation of actin filaments. Constitutively active Rac augmented cofilin phosphorylation and LIM-kinase 1 autophosphorylation whereas phorbol ester inhibited these processes. Our results define a mechanism for the regulation of cofilin and hence of actin dynamics in vivo. By modulating the stability of actin cytoskeletal structures, this pathway should play a central role in regulating cell motility and morphogenesis.
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- "In the last few decades, a large number of key molecules has been discovered and studied where they are understood to play an important role in the sensing of chemical stimuli as well as the subsequent polarization, regulation of the actin cytoskeleton and the generation of mechanical forces . Among these molecules are small GTPases  , PI3K, PTEN, PIPs,   , Arp2/3   and Cofilin  . "
ABSTRACT: During cell migration, cells become polarized, change their shape, and move in response to various cues, both internal and external. Many existing mathematical models of cell polarization are formulated in one or two spatial dimensions and hence cannot accurately capture the effect of cell shape, as well as the response of the cell to signals from different directions in a three-dimensional environment. To study those effects, we introduce a three-dimensional reaction-diffusion model of a cell. As some key molecules in cell polarization, such as the small GTPases, can exist both membrane bound and soluble in the cytosol, we first look at the role of cell geometry on the membrane binding/unbinding dynamics of such molecules. We derive quite general conditions under which effective existing one or two-dimensional computational models are valid, and find novel renormalizations of parameters in the effective model. We then extend an established one-dimensional cell polarization pathway in our three-dimensional framework. Our simulations indicate that even in some quasi-one-dimensional scenarios, such as polarization of a cell along a linear growth factor gradient, the cell shape can influence the polarization behavior of the cell, with cells of some shape polarizing more efficiently than those of other shapes. We also investigate the role of the previously ignored membrane unbinding rate on polarization. Furthermore, we simulate the response of the cell when the external signal is changing directions, and we find that more symmetric cells can change their polarized state more effectively towards the new stimulus than cells which are elongated along the direction of the original stimulus.
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- "Chai et al. (2009) have previously shown that Reelin phosphorylates the actindepolymerizing protein cofilin. Phosphorylation of cofilin renders it unable to depolymerize actin, thereby stabilizing the actin cytoskeleton (Arber et al. 1998; Yang et al. 1998). "
ABSTRACT: Newborn neurons migrate along the processes of radial glial cells (RGCs) to reach their final positions in the cortex. Here, we visualized individual migrating neurons and RGCs using in utero electroporation. We show that branching of migrating neurons and RGCs is closely correlated spatiotemporally with the distribution of Reelin. Time-lapse imaging revealed that the leading processes of migrating neurons gave rise to increasingly more branches once their growth cones contacted the Reelin-containing marginal zone. This was accompanied by translocation of the nucleus and gradual shortening of the leading process. Absence of Reelin in reeler mice altered these processes resulting in misorientation, loss of bipolarity, and aberrant migration of cortical neurons. Moreover, in reeler, the branching of the basal processes of RGCs in the marginal zone was severely disrupted. Consistent with previous reports, we show that in dissociated reeler cortical cultures, exposure to recombinant Reelin enhanced dendritic complexity and glial branching. Our results suggest that Reelin induces branching of the leading processes of migrating neurons and that of basal processes of RGCs when they arrive at the Reelin-containing marginal zone. Branching of these processes may be crucial for the termination of nuclear translocation during the migratory process and for correct neuronal positioning.Cerebral Cortex 09/2014; DOI:10.1093/cercor/bhu216 · 8.67 Impact Factor
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- "Changes in spine structure were accompanied by changes in the actin cytoskeleton and shifts in the equilibrium between F-actin and G-actin (Okamoto et al., 2004) by virtue of actin-depolymerizing proteins such as cofilin. The severing activity of cofilin is terminated by phosphorylation of the protein (Arber et al., 1998; Yang et al., 1998), and increased spine size in LTP was found to be associated with increased phosphorylation of cofilin (Chen et al., 2007; Rex et al., 2009), indicating stabilization of the spine cytoskeleton. "
ABSTRACT: Camillo Golgi’s “Reazione Nera” led to the discovery of dendritic spines, small appendages originating from dendritic shafts. With the advent of electron microscopy (EM) they were identified as sites of synaptic contact. Later it was found that changes in synaptic strength were associated with changes in the shape of dendritic spines. While live-cell imaging was advantageous in monitoring the time course of such changes in spine structure, EM is still the best method for the simultaneous visualization of all cellular components, including actual synaptic contacts, at high resolution. Immunogold labeling for EM reveals the precise localization of molecules in relation to synaptic structures. Previous EM studies of spines and synapses were performed in tissue subjected to aldehyde fixation and dehydration in ethanol, which is associated with protein denaturation and tissue shrinkage. It has remained an issue to what extent fine structural details are preserved when subjecting the tissue to these procedures. In the present review, we report recent studies on the fine structure of spines and synapses using high-pressure freezing (HPF), which avoids protein denaturation by aldehydes and results in an excellent preservation of ultrastructural detail. In these studies, HPF was used to monitor subtle fine-structural changes in spine shape associated with chemically induced long-term potentiation (cLTP) at identified hippocampal mossy fiber synapses. Changes in spine shape result from reorganization of the actin cytoskeleton. We report that cLTP was associated with decreased immunogold labeling for phosphorylated cofilin (p-cofilin), an actin-depolymerizing protein. Phosphorylation of cofilin renders it unable to depolymerize F-actin, which stabilizes the actin cytoskeleton. Decreased levels of p-cofilin, in turn, suggest increased actin turnover, possibly underlying the changes in spine shape associated with cLTP. The findings reviewed here establish HPF as an appropriate mFrontiers in Neuroanatomy 09/2014; 8. DOI:10.3389/fnana.2014.00094 · 3.54 Impact Factor