Chase-and-run between adjacent cell populations promotes directional collective migration

Collective cell migration in morphogenesis and cancer progression often involves the coordination of multiple cell types. How reciprocal interactions between adjacent cell populations lead to new emergent behaviours remains unknown. Here we studied the interaction between neural crest (NC) cells, a highly migratory cell population, and placodal cells, an epithelial tissue that contributes to sensory organs. We found that NC cells chase placodal cells by chemotaxis, and placodal cells run when contacted by NC. Chemotaxis to Sdf1 underlies the chase, and repulsion involving PCP and N-cadherin signalling is responsible for the run. This chase-and-run requires the generation of asymmetric forces, which depend on local inhibition of focal adhesions. The cell interactions described here are essential for correct NC migration and for segregation of placodes in vivo and are likely to represent a general mechanism of coordinated migration

Dimerization of the thyrotropin-releasing hormone receptor potentiates hormone-dependent receptor phosphorylation

The G protein-coupled thyrotropin (TSH)-releasing hormone (TRH) receptor forms homodimers. Regulated receptor dimerization increases TRH-induced receptor endocytosis. These studies test whether dimerization increases receptor phosphorylation, which could potentiate internalization. Phosphorylation at residues 355–365, which is critical for internalization, was measured with a highly selective phospho-site-specific antibody. Two strategies were used to drive receptor dimerization. Dimerization of a TRH receptor-FK506-binding protein (FKBP) fusion protein was stimulated by a dimeric FKBP ligand. The chemical dimerizer caused a large increase in TRH-dependent phosphorylation within 1 min, whereas a monomeric FKBP ligand had no effect. The dimerizer did not alter phoshorylation of receptors lacking the FKBP domain. Dimerization of receptors containing an N-terminal HA epitope also was induced with anti-HA antibody. Anti-HA IgG strongly increased TRH-induced phosphorylation, whereas monomeric Fab fragments had no effect. Anti-HA antibody did not alter phosphorylation in receptors lacking an HA tag. Furthermore, two phosphorylation-defective TRH receptors functionally complemented one another and permitted phosphorylation. Receptors with a D71A mutation in the second transmembrane domain do not signal, whereas receptors with four Ala mutations in the 355–365 region signal normally but lack phosphorylation sites. When D71A- and 4Ala-TRH receptors were expressed alone, neither underwent TRH-dependent phosphorylation. When they were expressed together, D71A receptor was phosphorylated by G protein-coupled receptor kinases in response to TRH. These results suggest that the TRH receptor is phosphorylated preferentially when it is in dimers or when preexisting receptor dimers are driven into microaggregates. Increased receptor phosphorylation may amplify desensitization

Bacteriorhodopsin Folds through a Poorly Organized Transition State

[Image: see text] The folding mechanisms of helical membrane proteins remain largely uncharted. Here we characterize the kinetics of bacteriorhodopsin folding and employ φ-value analysis to explore the folding transition state. First, we developed and confirmed a kinetic model that allowed us to assess the rate of folding from SDS-denatured bacteriorhodopsin (bR(U)) and provides accurate thermodynamic information even under influence of retinal hydrolysis. Next, we obtained reliable φ-values for 16 mutants of bacteriorhodopsin with good coverage across the protein. Every φ-value was less than 0.4, indicating the transition state is not uniquely structured. We suggest that the transition state is a loosely organized ensemble of conformations

In Vivo Analysis of the Mesenchymal-to-Epithelial Transition During Chick Secondary Neurulation

The neural tube in amniotic embryos forms as a result of two consecutive events along the anteroposterior axis, referred to as primary and secondary neurulation (PN and SN). While PN involves the invagination of a sheet of epithelial cells, SN shapes the caudal neural tube through the mesenchymal-to-epithelial transition (MET) of neuromesodermal progenitors, followed by cavitation of the medullary cord. The technical difficulties in studying SN mainly involve the challenge of labeling and manipulating SN cells in vivo. Here we describe a new method to follow MET during SN in the chick embryo, combining early in ovo chick electroporation with in vivo time-lapse imaging. This procedure allows the cells undergoing SN to be manipulated in order to investigate the MET process, permitting their cell dynamics to be followed in vivo

Involvement of a Golgi-resident GPI-anchored Protein in Maintenance of the Golgi Structure

The Golgi apparatus consists of a series of flattened cisternal membranes that are aligned in parallel to form stacks. Cytosolic-oriented Golgi-associated proteins have been identified that may coordinate or maintain the Golgi architecture. Here, we describe a novel GPI-anchored protein, Golgi-resident GPI-anchored protein (GREG) that has a brefeldin A-sensitive Golgi localization. GREG resides in the Golgi lumen as a cis-oriented homodimer, due to strong interactions between coiled-coil regions in the C termini. Dimerization of GREG as well as its Golgi localization depends on a unique tandem repeat sequence within the coiled-coil region. RNA-mediated interference of GREG expression or expression of GREG mutants reveals an essential role for GREG in maintenance of the Golgi integrity. Under these conditions, secretion of the vesicular stomatitis virus glycoprotein protein as a marker for protein transport along the secretory pathway is inhibited, suggesting a loss of Golgi function as well. These results imply the involvement of a luminal protein in Golgi structure and function

Neandertal spatial patterns and occupation dynamics: a regional focus on the central region in Mediterranean Iberia

En el siguiente trabajo se estudian varios conjuntos pertenecientes al Paleolítico medio procedentes del mediterráneo peninsular ibérico con el objetivo de examinar los patrones de ocupación y las estrategias de gestión del territorio. Se presta especial atención al abastecimiento de las materias primas y los comportamientos tecnológicos, los datos procedentes de la fauna y los análisis microespaciales. La variabilidad en los tipos de ocupación de los distintos conjuntos nos muestra una gran diversidad y una multitud de factores, aunque no parece tener una sola explicación cultural, funcional, temporal o ambiental. Más bien son explicaciones que responden a una amplia variabilidad en los comportamientos técnicos observados y que se explican en función de las propias necesidades de las poblaciones dentro de cada región. Los resultados obtenidos nos permiten analizar los datos y compararlos en el contexto del sudoeste de Europa de cara a elaborar un primer enfoque de las estrategias de subsistencia de los neandertales y su movilidad en una región hasta ahora poco conocida desde este punto de vista.HAR2017-85,153-P/MICINNPROMETEO/2017/060HAR2016-76,760-C3-1-P/MICINNThis paper focuses on the study of some Middle Palaeolithic assemblages from Mediterranean Iberia to examine Neanderthal occupation patterns and territory management strategies, paying special attention to raw material procurement and technological behaviours, zooarchaeological data and microspatial patterning. The site occupation types are variable, and some of the results may have more importance than is immediately apparent, but there does not seem to be a single cultural, functional, temporal or environmental explanation. Rather, the wide variability in the technical behaviours observed can be explained with reference to the particular requirements of the populations in each specific region. The results obtained allow us to interrogate the data and, drawing comparisons with the southwest European context, establish an initial approach to Neanderthal subsistence strategies and mobility in a region so far little known in this regard