Tyrosine Kinase-Dependent Activation of Phospholipase Cγ Is Required for Calcium Transient in Xenopus Egg Fertilization

AbstractIn a previous study (K.-I. Sato et al., 1999, Dev. Biol. 209, 308–320), we presented evidence that a Src-related protein-tyrosine kinase (PTK), named Xyk, may act upstream of the calcium release in fertilization of the Xenopus egg. In the present study, we examined whether PTK activation of phospholipase Cγ (PLCγ) plays a role in the fertilization-induced calcium signaling. Immunoprecipitation studies show that Xenopus egg PLCγ is tyrosine phosphorylated and activated within a few minutes after fertilization but not after A23187-induced egg activation. Consistently, we observed a fertilization-induced association of PLCγ with Xyk activity that was not seen in A23187-activated eggs. A Src-specific PTK inhibitor, PP1, blocked effectively the fertilization-induced association of PLCγ with Xyk activity and up-regulation of PLCγ, when microinjected into the egg. In addition, a PLC inhibitor, U-73122, inhibited sperm-induced inositol 1,4,5-trisphosphate production and the calcium transient and subsequent calcium-dependent events such as cortical contraction, elevation of fertilization envelope, and tyrosine dephosphorylation of p42 MAP kinase, all of which were also inhibited by PP1. On the other hand, A23187 could cause the calcium response and calcium-dependent events in eggs injected with PP1 or U-73122. These results support the idea that Xenopus egg fertilization requires Src-family PTK-dependent PLCγ activity that acts upstream of the calcium-dependent signaling pathway

Evidence that phosphatidylinositol 3-kinase is involved in sperm-induced tyrosine kinase signaling in Xenopus egg fertilization

<p>Abstract</p> <p>Background</p> <p>Studies have examined the function of PI 3-kinase in the early developmental processes that operate in oocytes or early embryos of various species. However, the roles of egg-associated PI 3-kinase and Akt, especially in signal transduction at fertilization, are not well understood.</p> <p>Results</p> <p>Here we show that in <it>Xenopus </it>eggs, a potent inhibitor of phosphatidylinositol 3-kinase (PI 3-kinase), LY294002 inhibits sperm-induced activation of the tyrosine kinase Src and a transient increase in the intracellular concentration of Ca²⁺at fertilization. LY294002 also inhibits sperm-induced dephosphorylation of mitogen-activated protein kinase, breakdown of cyclin B2 and Mos, and first embryonic cleavage, all of which are events of Ca²⁺-dependent egg activation. In fertilized eggs, an 85-kDa subunit of PI 3-kinase (p85) undergoes a transient translocation to the low-density, detergent-insoluble membranes (membrane microdomains) where Src tyrosine kinase signaling is operating. However, the tyrosine phosphorylation of p85 in fertilized eggs is not as evident as that in H2O2-activated eggs, arguing against the possibility that PI 3-kinase is activated by Src phosphorylation. Nevertheless, sperm-induced activation of PI 3-kinase has been demonstrated by the finding that Akt, a serine/threonine-specific protein kinase, is phosphorylated at threonine-308. The threonine-phosphorylated Akt also localizes to the membrane microdomains of fertilized eggs. Application of bp(V), an inhibitor of PTEN that dephosphorylates PIP3, the enzymatic product of PI 3-kinase, promotes parthenogenetic activation of <it>Xenopus </it>eggs. In vitro kinase assays demonstrate that PIP3 activates Src in a dose-dependent manner.</p> <p>Conclusions</p> <p>These results suggest that PI 3-kinase is involved in sperm-induced egg activation via production of PIP3 that would act as a positive regulator of the Src signaling pathway in <it>Xenopus </it>fertilization.</p

Evidence for the Involvement of a Src-Related Tyrosine Kinase inXenopusEgg Activation

AbstractRecently, we have purified a Src-related tyrosine kinase, namedXenopustyrosine kinase (Xyk), from oocytes ofXenopus laevisand found that the enzyme is activated within 1 min following fertilization [Satoet al.(1996)J. Biol. Chem.271, 13250–13257]. A concomitant translocation of a part of the activated enzyme from the membrane fraction to the cytosolic fraction was also observed. In the present study, we show that parthenogenetic egg activation by a synthetic RGDS peptide [Y. Iwao and T. Fujimura, T. (1996)Dev. Biol.177, 558–567], an integrin-interacting peptide, but not by electrical shock or the calcium ionophore A23187 causes the kinase activation, tyrosine phosphorylation, and translocation of Xyk. A synthetic tyrosine kinase-specific inhibitor peptide was employed to analyze the importance of the Xyk activity in egg activation. We found that the peptide inhibits the kinase activity of purified Xyk at IC50of 8 μM. Further, egg activation induced by sperm or RGDS peptide but not by A23187 was inhibited by microinjection of the peptide. In the peptide-microinjected eggs, penetration of the sperm nucleus into the egg cytoplasm and meiotic resumption in the egg were blocked. Indirect immunofluorescence study demonstrates that Xyk is exclusively localized to the cortex ofXenopuseggs, indicating that Xyk can function in close proximity to the sperm–egg or RGDS peptide–egg interaction site. Taken together, these data suggest that the tyrosine kinase Xyk plays an important role in the early events ofXenopusegg activation in a manner independent or upstream of calcium signaling

The Quark-Hadron Phase Transition, QCD Lattice Calculations and Inhomogeneous Big-Bang Nucleosynthesis

We review recent lattice QCD results for the surface tension at the finite temperature quark-hadron phase transition and discuss their implications on the possible scale of inhomogeneities. In the quenched approximation the average distance between nucleating centers is smaller than the diffusion length of a protron, so that inhomogeneities are washed out by the time nucleosynthesis sets in. Consequently the baryon density fluctuations formed by a QCD phase transition in the early universe cannot significantly affect standard big-bang nucleosynthesis calculations and certainly cannot allow baryons to close the universe. At present lattice results are inconclusive when dynamical fermions are included.Comment: 8 pages, LaTe

Precision spectroscopy of pionic 1s states of Sn nuclei and evidence for partial restoration of chiral symmetry in the nuclear medium

Deeply bound 1s states of $\pi^-$ in $^{115,119,123}$Sn were preferentially observed using the Sn($d$,$^3$He) pion-transfer reaction under the recoil-free condition. The 1s binding energies and widths were precisely determined, and were used to deduce the isovector parameter of the s-wave pion-nucleus potential to be $b_1 =-0.115\pm 0.007 ~m_{\pi}^{-1}$. The observed enhancement of $|b_1|$ over the free $\pi N$ value ($b_1^{\rm free}/b_1 = 0.78 \pm 0.05$) indicates a reduction of the chiral order parameter, $f^{*}_{\pi} (\rho)^2/f_{\pi}^2 \approx 0.64$, at the normal nuclear density, $\rho = \rho_0$.Comment: 4 pages including 3 postscript figures, RevTeX 4 with multirow.sty, submitted to Physical Review Letter

Skyrmion Hall Effect Revealed by Direct Time-Resolved X-Ray Microscopy

Magnetic skyrmions are highly promising candidates for future spintronic applications such as skyrmion racetrack memories and logic devices. They exhibit exotic and complex dynamics governed by topology and are less influenced by defects, such as edge roughness, than conventionally used domain walls. In particular, their finite topological charge leads to a predicted "skyrmion Hall effect", in which current-driven skyrmions acquire a transverse velocity component analogous to charged particles in the conventional Hall effect. Here, we present nanoscale pump-probe imaging that for the first time reveals the real-time dynamics of skyrmions driven by current-induced spin orbit torque (SOT). We find that skyrmions move at a well-defined angle {\Theta}_{SH} that can exceed 30{\deg} with respect to the current flow, but in contrast to theoretical expectations, {\Theta}_{SH} increases linearly with velocity up to at least 100 m/s. We explain our observation based on internal mode excitations in combination with a field-like SOT, showing that one must go beyond the usual rigid skyrmion description to unravel the dynamics.Comment: pdf document arxiv_v1.1. 24 pages (incl. 9 figures and supplementary information

Search for the Θ+\Theta^{+} pentaquark via the π−p→K−X\pi^-p\to K^-X reaction at 1.92 GeV/cc

The $\Theta^+$ pentaquark baryon was searched for via the $\pi^-p\to K^-X$ reaction in a missing-mass resolution of 1.4 MeV/$c^2$(FWHM) at J-PARC. $\pi^-$ meson beams were incident on the liquid hydrogen target with the beam momentum of 1.92 GeV/$c$. No peak structure corresponding to the $\Theta^+$ mass was observed. The upper limit of the production cross section averaged over the scattering angle of 2$^{\circ}$ to 15$^{\circ}$ in the laboratory frame was obtained to be 0.26 $\mu$b/sr in the mass region of 1.51$-$1.55 GeV/$c^2$.The upper limit of the $\Theta^+$ decay width using the effective Lagrangian approach was obtained to be 0.72 MeV/$c^2$ and 3.1 MeV/$c^2$ for $J^P_{\Theta}=1/2^+$ and $J^P_{\Theta}=1/2^-$, respectively.Comment: 5 pages, 3 figures, 1 tabl

Water waves generated by a moving bottom

Tsunamis are often generated by a moving sea bottom. This paper deals with the case where the tsunami source is an earthquake. The linearized water-wave equations are solved analytically for various sea bottom motions. Numerical results based on the analytical solutions are shown for the free-surface profiles, the horizontal and vertical velocities as well as the bottom pressure.Comment: 41 pages, 13 figures. Accepted for publication in a book: "Tsunami and Nonlinear Waves", Kundu, Anjan (Editor), Springer 2007, Approx. 325 p., 170 illus., Hardcover, ISBN: 978-3-540-71255-8, available: May 200

Observation of B+->Lambda_c+Lambda_c-K+ and B0->Lambda_c+Lambda_c^-K0 decays

We report first measurements of the doubly charmed baryonic B decays B->Lambda_c+Lambda_c-K. The B+->Lambda_c+Lambda_c-K+ decay is observed with a branching fraction of (6.5+1.0-0.9+-1.1+-3.4)x10^{-4} and a statistical significance of 15.4sigma. The B0->Lambda_c+Lambda_c-K0 decay is observed with a branching fraction of (7.9+2.9-2.3+-1.2+-4.2)x10^{-4} and a statistical significance of 6.6sigma. The branching fraction errors are statistical, systematic, and the error resulting from the uncertainty of Lambda_c+->pK-pi+ decay branching fraction. The analysis is based on 357fb-1 of data accumulated at the Y(4S) resonance with the Belle detector at the KEKB asymmetric e+e- collider.Comment: 10 pages, 3 figure