Masatoshi Takeichi

Morphogenetic roles of classic cadherins

Classic cadherins, which are known to be crucial for homotypic cell-cell adhesion, have been found to be present not only in vertebrate but also in invertebrate species. Their three-dimensional structures, novel functions, and novel expression patterns were reported recently. These have been important steps towards a deeper understanding of the morphogenetic roles of this family of molecules.

Cadherins in cancer: implications for invasion and metastasis.

In order to understand the mechanisms of metastasis, one must first determine how cancer cells detach from primary tumors. It is known that cadherins are major cell-cell adhesion molecules in tumors as well as in normal tissues. The perturbation of cadherin function causes temporal or permanent disaggregation of tumor cells and may thus promote the invasion and metastasis of such cells.

Flamingo, a Seven-Pass Transmembrane Cadherin, Regulates Planar Cell Polarity under the Control of Frizzled

We identified a seven-pass transmembrane receptor of the cadherin superfamily, designated Flamingo (Fmi), localized at cell-cell boundaries in the Drosophila wing. In the absence of Fmi, planar polarity was distorted. Before morphological polarization of wing cells along the proximal-distal (P-D) axis, Fmi was redistributed predominantly to proximal and distal cell edges. This biased localization of Fmi appears to be driven by an imbalance of the activity of Frizzled (Fz) across the proximal/distal cell boundary. These results, together with phenotypes caused by ectopic expression of fz and fmi, suggest that cells acquire the P-D polarity by way of the Fz-dependent boundary localization of Fmi.

Expressed recombinant cadherins mediate cell sorting in model systems

Cadherins are cell-surface glycoproteins responsible for Ca2+-dependent cell-to-cell adhesion. E- or P-cadherin was transfected into L cells, which normally have little cadherin activity, and cellular aggregation of the resulting transfectants was observed to be a function of the cadherin molecule expressed. Transfected cells preferentially adhered to cells expressing the same cadherin subclass. Furthermore, in reconstituted embryonic lung tissue, E-cadherin-expressing L cells were associated with epithelial tubules expressing E-cadherin, while untransfected L cells associated with mesenchymal cells. These results provide the first direct evidence that the differential expression of cadherins can play a role in cell sorting in heterogeneous cell populations.

Control of Actin Reorganization by Slingshot, a Family of Phosphatases that Dephosphorylate ADF/Cofilin

The ADF (actin-depolymerizing factor)/cofilin family is a stimulus-responsive mediator of actin dynamics. In contrast to the mechanisms of inactivation of ADF/cofilin by kinases such as LIM-kinase 1 (LIMK1), much less is known about its reactivation through dephosphorylation. Here we report Slingshot (SSH), a family of phosphatases that have the property of F actin binding. In Drosophila, loss of ssh function dramatically increased levels of both F actin and phospho-cofilin (P cofilin) and disorganized epidermal cell morphogenesis. In mammalian cells, human SSH homologs (hSSHs) suppressed LIMK1-induced actin reorganization. Furthermore, SSH and the hSSHs dephosphorylated P cofilin in cultured cells and in cell-free assays. Our results strongly suggest that the SSH family plays a pivotal role in actin dynamics by reactivating ADF/cofilin in vivo.

Identification of a neural α-catenin as a key regulator of cadherin function and multicellular organization

The function of cadherin cell adhesion molecules is thought to be regulated by a group of cytoplasmic proteins, including alpha-catenin. We identified a subtype of alpha-catenin, termed alpha N-catenin, which is associated with N-cadherin and expressed mainly in the nervous system. cDNA transfection experiments showed that alpha N-catenin can also bind with E-cadherin. To investigate the role of alpha N-catenin, we transfected lung carcinoma PC9 cells, which express E-cadherin and beta-catenin but neither alpha- nor alpha N-catenin, with alpha N-catenin cDNA. While parental PC9 grew as isolated cells, the transfectant lines formed aggregates in which cells were tightly adhered to each other, showing epithelial arrangements, and they occasionally gave rise to cystic spheres. These results suggest that alpha N-catenin is crucial not only for cadherin function but also for organization of multicellular structures.

Localization of specificity determining sites in cadherin cell adhesion molecules

Cadherins are a group of homophilic intercellular adhesion molecules; each member of this family exhibits binding specificity. Here, we attempted to map the sites for the specificities of these molecules by analyzing adhesives selectivities of the cells that express chimeric and point-mutated E- and P-cadherin. The results showed that the amino-terminal 113 amino acid region is essential to determine the specificities, and within this region we could identify especially important sites in which amino acid substitutions altered the binding specificity of cadherins. We also found that the epitopes for antibodies capable of blocking cadherin action are located in this amino-terminal region.

Spatial and temporal expression pattern of N-cadherin cell adhesion molecules correlated with morphogenetic processes of chicken embryos☆

N-cadherin is a Ca2+-dependent cell-cell adhesion molecule, which was identified in brain cells of mouse and chicken. In the present study, we have determined the pattern of expression of N-cadherin in chicken embryos at various stages by means of immunohistochemistry. N-cadherin was expressed in cells derived from all three primary germ layers. Its expression was transient in many tissues but permanent in others. The transient expression occurred in nephric tubules, skeletal muscles, mesenchymal tissues, endodermal organs, and epidermis, while the permanent expression occurred in nervous systems, lens, and myocardiac cells. Appearance or disappearance of N-cadherin could be generally correlated with morphogenetic events, such as rearrangement, segregation, or association of cells. Comparison of the expression pattern of N-cadherin with that of L-CAM and N-CAM determined by other workers suggests that there is some mechanism controlling expression of multiple classes of adhesion molecules. The pattern of expression of N-cadherin was generally complementary to that of L-CAM; that is, if N-cadherin appeared, L-CAM disappeared or vice versa. We also found cases in which N-cadherin was expressed in the same local regions as L-CAM. The distribution of N-cadherin was similar to that of N-CAM with some exceptions. Thus, N-cadherin and other cell-cell adhesion molecules seem to be expressed under a precise spatial and temporal control so as to be associated with a variety of morphogenetic events during development.

The 102 kd cadherin-associated protein: Similarity to vinculin and posttranscriptional regulation of expression

The E-cadherin cell adhesion molecule is associated with cytoplasmic polypeptides, and this association is essential for its cell-binding function. Using isolated adherens junctions of the liver, we purified a 102 kd protein that can associate with E-cadherin (CAP102) and isolated cDNAs encoding this protein. Sequence analysis of the cDNAs revealed that this protein has a similarity to vinculin. L cells not expressing endogenous cadherin express the mRNA for CAP102 but have only a trace amount of CAP102 protein. Introducing exogenous E-cadherin into these cells, however, induced a high expression of CAP102 protein without affecting the amount of its mRNA, suggesting that there is a posttranscriptional regulatory mechanism for this molecule. The same effect was observed by introducing N- or P-cadherin into L cells.

Cadherin Regulates Dendritic Spine Morphogenesis

Synaptic remodeling has been postulated as a mechanism underlying synaptic plasticity, and cadherin adhesion molecules are thought to be a regulator of such a process. We examined the effects of cadherin blockage on synaptogenesis in cultured hippocampal neurons. This blockade resulted in alterations of dendritic spine morphology, such as filopodia-like elongation of the spine and bifurcation of its head structure, along with concomitant disruption of the distribution of postsynaptic proteins. The accumulation of synapsin at presynaptic sites and synaptic vesicle recycling were also perturbed, although these synaptic responses to the cadherin blockade became less evident upon the maturation of the synapses. These findings suggest that cadherin regulates dendritic spine morphogenesis and related synaptic functions, presumably cooperating with cadherin-independent adhesive mechanisms to maintain spine-axon contacts.