Dephosphorylation of Y228 and Y217 and phosphorylation of Y335 in p120 catenin activate convergent extension during zebrafish gastrulation.

BACKGROUND: The cadherin-associated protein p120 catenin regulates convergent extension through interactions with cadherin proteins, Cdc42, and Rac1, as we previously showed in zebrafish (Danio rerio). Phosphorylation of p120 catenin changes the nature of its activity in vitro but is virtually unexplored in embryos. We used our previously developed antisense RNA splice-site morpholino targeted to endogenous p120 catenin-δ1 to cause defects in axis elongation probing the functions of three p120 catenin tyrosine-phosphorylation sites in gastrulating zebrafish embryos. RESULTS: The morpholino-induced defects were rescued by co-injections with mouse p120 catenin-δ1-3A mRNAs mutated at residues Y228 and Y217 to a non-phosphorylatable phenylalanine (F) or mutated at residue Y335 to a phosphomimetic glutamic acid (E). Co-injection of the complementary mutations Y228E, Y217E, or Y335F mRNAs partially rescued embryos whereas dual mutation to Y228E-Y217E blocked rescue. Immunopurification showed Y228F mutant proteins preferentially interacted with Rac1, potentially promoting cell migration. In contrast, the phosphomimetic Y228E preferentially interacted with E-cadherin increasing adhesion. Both Y228F and Y335F strongly bind VAV2. CONCLUSIONS: p120 catenin serves dual roles during gastrulation of zebrafish. Phosphorylation and dephosphorylation of tyrosine residues Y217, Y228, and Y335 precisely balance cell adhesion and cell migration to facilitate somite compaction and axis elongation.

Phosphorylation of serine residues S252, S268/S269, and S879 in p120 catenin activates migration of presomitic mesoderm in gastrulating zebrafish embryos.

BACKGROUND: Cadherin-associated protein p120 catenin regulates cell adhesion and migration in cell cultures and is required for axial elongation in embryos. Its roles in adhesion and cell migration are regulated by phosphorylation. We determined the effects of phosphorylation of six serine and three threonine residues in p120 catenin during zebrafish (Danio rerio) embryogenesis. RESULTS: We knocked down endogenous p120 catenin-δ1 with an antisense RNA-splice-site morpholino (Sp-MO) causing defects in axis elongation. These defects were rescued by co-injections of mRNAs for wildtype mouse p120 catenin-δ1-3A or various mutated forms. Several mRNAs containing serine or threonine codons singly or doubly mutated to phosphomimetic glutamic acid rescued, and some nonphosphorylatable mutants did not. CONCLUSIONS: We discovered that phosphorylation of serine residue S252 or S879 is required for convergent extension of zebrafish embryos, since rescue occurred only when these residues were mutated to glutamic acid. In addition, the phosphorylation of either S268 or S269 is required, not both, consistent with the presence of only a single one of these residues in two isoforms of zebrafish and Xenopus laevis. In summary, phosphorylation of multiple serine and threonine residues of p120 catenin activates migration of presomitic mesoderm of zebrafish embryos facilitating elongation of the dorsal axis.

Electronic spectroscopy of Mg-phthalocyanine embedded in cold hydrogen clusters produced by a pulsed nozzle.

Cold clusters of molecular hydrogen were created using a pulsed nozzle. The thermodynamical states of the clusters were characterized by measuring the cluster beam velocity and the laser-induced fluorescence (LIF) spectra of embedded molecules. Two distinct velocity components were identified in the beam that originates from different clustering mechanisms. The fast velocity component corresponds to the expansion of H2 from the gas phase, while the slow velocity component corresponds to the expansion from the liquid phase. The velocity distribution of these two components showed no significant difference between the expansions of para and normal hydrogen. In this study, LIF spectroscopy of single Mg-phthalocyanine molecules embedded in the H2 clusters consisting of 105 H2 molecules was used to investigate the properties of the fast component. The observed peak frequencies of the LIF signals, compared to those observed in helium droplets, were used to infer the possible presence of the liquid phase in the fast component of the H2 clusters below 5 K. The shift, linewidth, and splitting in the spectra, which strongly depend on the ortho/para ratio, are attributed to the local configurations of hydrogen in the vicinity of the probe molecules.

Design and characterization of a novel toroidal split-ring resonator.

The design and characterization of a novel toroidal split-ring resonator (SRR) are described in detail. In conventional cylindrical SRRs, there is a large magnetic flux within the bore of the resonator. However, there also exists a non-negligible magnetic flux in the free space surrounding the resonator. The energy losses associated with this radiated power diminish the resonator's quality factor. In the toroidal SRR, on the other hand, the magnetic field lines are strongly confined within the bore of the resonator resulting in high intrinsic quality factors and stable resonance frequencies without requiring additional electromagnetic shielding. This paper describes the design and construction of a toroidal SRR as well as an experimental investigation of its cw response in the frequency-domain and its time-domain response to a rf pulse. Additionally, the dependence of the toroidal SRR's resonant frequency and quality factor on the strength of inductive coupling to external circuits is investigated both theoretically and experimentally.

Laser induced fluorescence spectroscopy of tetracene with large Ar, Ne, and H2 clusters in superfluid He nanodroplets.

Clusters of tetracene molecules with different numbers of attached (Ar)(N), (Ne)(N) and (H(2))(N) particles (N = 1-2000) are assembled inside superfluid He nanodroplets and studied via laser-induced fluorescence. The frequency shift of the fluorescence spectrum of the tetracene molecules is studied as a function of cluster size and pickup order of tetracene and cluster species. For (Ar)(N) and (Ne)(N) clusters, our results indicate that the tetracene molecules reside inside the clusters when tetracene is captured by the He nanodroplet before the cluster species; conversely, the tetracene molecules stay on the surface of the clusters when tetracene is captured after the cluster species. In the case of (H(2))(N) clusters, however, tetracene molecules reside inside the (H(2))(N) clusters irrespective of the pickup order. We conclude that (Ar)(N) and (Ne)(N) clusters are rigid at T = 0.38 K, while (H(2))(N) clusters of up to N = 2000 remain fluxional at the same temperature. The results may also indicate the occurrence of heterogeneous nucleation of the (H(2))(N) clusters, which is induced by the interaction with tetracene chromophore molecules.

A food-derived synergist of NGF signaling: identification of protein tyrosine phosphatase 1B as a key regulator of NGF receptor-initiated signal transduction.

Neurotrophins, such as the nerve growth factor (NGF), play an essential role in the growth, development, survival and functional maintenance of neurons in the central and peripheral systems. They also prevent neuronal cell death under various stressful conditions, such as ischemia and neurodegenerative disorders. NGF induces cell differentiation and neurite outgrowth by binding with and activating the NGF receptor tyrosine kinase followed by activation of a variety of signaling cascades. We have investigated the NGF-dependent neuritogenesis enhancer potential of a food-derived small molecule contained in Brassica vegetables and identified the protein tyrosine phosphatase (PTP) 1B as a key regulator of the NGF receptor-initiated signal transduction. Based on an extensive screening of Brassica vegetable extracts for the neuritogenic-promoting activity in the rat pheochromocytoma cell line PC12, we found the Japanese horseradish, wasabi (Wasabia japonica, syn. Eutrema wasabi), as the richest source and identified 6-methylsulfinylhexyl isothiocyanate (6-HITC), an analogue of sulforaphane isolated from broccoli, as one of the major neuritogenic enhancers in the wasabi. 6-HITC strongly enhanced the neurite outgrowth and neurofilament expression elicited by a low-concentration of NGF that alone was insufficient to induce neuronal differentiation. 6-HITC also facilitated the sustained-phosphorylation of the extracellular signal-regulated kinase and the autophosphorylation of the NGF receptor TrkA. It was found that PTP1B act as a phosphatase capable of dephosphorylating Tyr-490 of TrkA and was inactivated by 6-HITC in a redox-dependent manner. The identification of PTP1B as a regulator of NGF signaling may provide new clues about the chemoprotective potential of food components, such as isothiocyanates.

Polymer-assisted solution-phase synthesis and neurite-outgrowth-promoting activity of 15-deoxy-delta(12,14)-PGJ2 derivatives.

An efficient solution-phase synthesis of rac-15-deoxy-delta(12,14)-PGJ2 (15dPGJ2) derivatives that contain variable alpha and omega chains based on a polymer-assisted strategy and their neurite-outgrowth-promoting activity are described. The strategy for the synthesis of PGJ2 derivatives involves the use of a vinyl iodide bearing cyclopentenone as a key intermediate, which undergoes Suzuki-Miyaura coupling and subsequent Lewis acid catalyzed aldol condensation for incorporation of the omega and alpha chains, respectively. For easy access to the PGJ2 derivatives, a polymer-supported catalyst and scavengers were adapted for use in these four diverse steps, in which workup and purification can be performed by simple filtration of the solid-supported reagents. By using this methodology, we succeeded in the synthesis of 16 PGJ2 derivatives with four alkyl boranes and four aldehydes. The neurite-outgrowth-promoting activity of the 16 synthetic compounds in PC12 cells revealed that the side-chains play a major role in modulating their biological activity. The carboxylic acid on the alpha chain improved the biological activity, although it was not absolutely required. Furthermore, a PGJ2 derivative with a phenyl moiety on the omega chain was found to exhibit an activity comparable to that of natural 15dPGJ2.

Neuromagnetic evidence that gingiva area is adjacent to tongue area in human primary somatosensory cortex.

The somatotopic organization of the human primary somatosensory (SI) area in the cerebral cortex has been intensively studied for the hand, lip, and tongue, but little is known about the gingiva. Penfield concluded that the gingival SI area was above the tongue area, as shown in his famous homunculus map. However, our recent study suggested that the lingual gingiva area was not so different to the tongue area. To delineate the fine SI somatotopy of the gingiva area, evoked magnetic fields were measured in 6 healthy subjects for the stimulus of the anterior or posterior and upper or lower parts of the lip, buccal and lingual gingiva, and tongue. Source position was estimated by a current dipole model at the first peak of the posterior-oriented current in a total of 12 cerebral hemispheres contralateral to the stimulation side. No significant difference was found between the positions of anterior and posterior or upper and lower parts of each structure. Both buccal and lingual gingiva areas were localized adjacent to the tongue area, but significantly lower than the lip area. We believe that the fine SI somatotopy of the human oral structures should be reconsidered.

Biomechanical comparison of straight and staggered implant placement configurations.

The effect of buccolingual staggered implant placement on stress distribution within the supporting structure was examined photoelastically. Two photoelastic models of a human mandible, edentulous distal to the canine, were fabricated. Three screw-type implants were embedded into the edentulous region of each model. The implants were placed in a straight line in one model and in a buccolingual staggered configuration in the other. Vertical and lateral loads were applied to a fixed partial denture superstructure. No clear biomechanical advantage to a staggered 1.5 mm buccal and lingual offset placement configuration was observed.