Spindle checkpoint proteins Mad1 and Mad2 are required for cytostatic factor–mediated metaphase arrest

In cells containing disrupted spindles, the spindle assembly checkpoint arrests the cell cycle in metaphase. The budding uninhibited by benzimidazole (Bub) 1, mitotic arrest-deficient (Mad) 1, and Mad2 proteins promote this checkpoint through sustained inhibition of the anaphase-promoting complex/cyclosome. Vertebrate oocytes undergoing meiotic maturation arrest in metaphase of meiosis II due to a cytoplasmic activity termed cytostatic factor (CSF), which appears not to be regulated by spindle dynamics. Here, we show that microinjection of Mad1 or Mad2 protein into early Xenopus laevis embryos causes metaphase arrest like that caused by Mos. Microinjection of antibodies to either Mad1 or Mad2 into maturing oocytes blocks the establishment of CSF arrest in meiosis II, and immunodepletion of either protein blocked the establishment of CSF arrest by Mos in egg extracts. A Mad2 mutant unable to oligomerize (Mad2 R133A) did not cause cell cycle arrest in blastomeres or in egg extracts. Once CSF arrest has been established, maintenance of metaphase arrest requires Mad1, but not Mad2 or Bub1. These results suggest a model in which CSF arrest by Mos is mediated by the Mad1 and Mad2 proteins in a manner distinct from the spindle checkpoint

Altered hippocampal function in major depression despite intact structure and resting perfusion

Background: Hippocampal volume reductions in major depression have been frequently reported. However, evidence for functional abnormalities in the same region in depression has been less clear. We investigated hippocampal function in depression using functional magnetic resonance imaging (fMRI) and neuropsychological tasks tapping spatial memory function, with complementing measures of hippocampal volume and resting blood flow to aid interpretation. Method: A total of 20 patients with major depressive disorder (MDD) and a matched group of 20 healthy individuals participated. Participants underwent multimodal magnetic resonance imaging (MRI): fMRI during a spatial memory task, and structural MRI and resting blood flow measurements of the hippocampal region using arterial spin labelling. An offline battery of neuropsychological tests, including several measures of spatial memory, was also completed. Results: The fMRI analysis showed significant group differences in bilateral anterior regions of the hippocampus. While control participants showed task-dependent differences in blood oxygen level-dependent (BOLD) signal, depressed patients did not. No group differences were detected with regard to hippocampal volume or resting blood flow. Patients showed reduced performance in several offline neuropsychological measures. All group differences were independent of differences in hippocampal volume and hippocampal blood flow. Conclusions: Functional abnormalities of the hippocampus can be observed in patients with MDD even when the volume and resting perfusion in the same region appear normal. This suggests that changes in hippocampal function can be observed independently of structural abnormalities of the hippocampus in depression

Arc Statistics in Clusters: Galaxy Contribution

The frequency with which background galaxies appear as long arcs as a result of gravitational lensing by foreground clusters of galaxies has recently been found to be a very sensitive probe of cosmological models by Bartelmann et al. (1998). They have found that such arcs would be expected far less frequently than observed (by an order of magnitude) in the currently favored model for the universe, with a large cosmological constant $\Omega_\Lambda \sim 0.7$. Here we analyze whether including the effect of cluster galaxies on the likelihood of clusters to generate long-arc images of background galaxies can change the statistics. Taking into account a variety of constraints on the properties of cluster galaxies, we find that there are not enough sufficiently massive galaxies in a cluster for them to significantly enhance the cross section of clusters to generate long arcs. We find that cluster galaxies typically enhance the cross section by only $\lesssim 15$.Comment: 19 pages, 1 figure, uses aasms4.sty, submitted to Ap

Bub1 is activated by the protein kinase p90Rsk during Xenopus oocyte maturation

AbstractBackground: The kinetochore attachment (spindle assembly) checkpoint arrests cells in metaphase to prevent exit from mitosis until all the chromosomes are aligned properly at the metaphase plate. The checkpoint operates by preventing activation of the anaphase-promoting complex (APC), which triggers anaphase by degrading mitotic cyclins and other proteins. This checkpoint is active during normal mitosis and upon experimental disruption of the mitotic spindle. In yeast, the serine/threonine protein kinase Bub1 and the WD-repeat protein Bub3 are elements of a signal transduction cascade that regulates the kinetochore attachment checkpoint. In mammalian cells, activated MAPK is present on kinetochores during mitosis and activity is upregulated by the spindle assembly checkpoint. In vertebrate unfertilized eggs, a special form of meiotic metaphase arrest by cytostatic factor (CSF) is mediated by MAPK activation of the protein kinase p90Rsk, which leads to inhibition of the APC. However, it is not known whether CSF-dependent metaphase arrest caused by p90Rsk involves components of the spindle assembly checkpoint.Results: xBub1 is present in resting oocytes and its protein level increases slightly during oocyte maturation and early embryogenesis. In Xenopus oocytes, Bub1 is localized to kinetochores during both meiosis I and meiosis II, and the electrophoretic mobility of Bub1 upon SDS-PAGE decreases during meiosis I, reflecting phosphorylation and activation of the enzyme. The activation of Bub1 can be induced in interphase egg extracts by selective stimulation of the MAPK pathway by c-Mos, a MAPKKK. In oocytes treated with the MEK1 inhibitor U0126, the MAPK pathway does not become activated, and Bub1 remains in its low-activity, unshifted form. Injection of a constitutively active target of MAPK, the protein kinase p90Rsk, restores the activation of Bub1 in the presence of U0126. Moreover, purified p90Rsk phosphorylates Bub1 in vitro and increases its protein kinase activity.Conclusions: Bub1, an upstream component of the kinetochore attachment checkpoint, is activated during meiosis in Xenopus in a MAPK-dependent manner. Moreover, a single substrate of MAPK, p90Rsk, is sufficient to activate Bub1 in vitro and in vivo. These results indicate that in vertebrate eggs, kinetochore attachment/spindle assembly checkpoint proteins, including Bub1, are downstream of p90Rsk and may be effectors of APC inhibition and CSF-dependent metaphase arrest by p90Rsk

Activation of p34cdc2 kinase by cyclin A

Functional clam cyclin A and B proteins have been produced using a baculovirus expression system. Both cyclin A and B can induce meiosis I and meiosis II in Xenopus in the absence of protein synthesis. Half-maximal induction occurs at 50 nM for cyclin A and 250 nM for cyclin B. Addition of 25 nM cyclin A to activated Xenopus egg extracts arrested in the cell cycle by treatment with RNase or emetine activates cdc2 kinase to the normal metaphase level and stimulates one oscillatory cell cycle. High levels of cyclin A cause marked hyperactivation of cdc2 kinase and a stable arrest at the metaphase point in the cell cycle. Kinetic studies demonstrate the concentration of cyclin A added does not affect the 10 min lag period required for kinase activation or the timing of maximal activity, but does control the rate of deactivation of cdc2 kinase during exit from mitosis. In addition, exogenous clam cyclin A inhibits the degradation of both A- and B-type endogenous Xenopus cyclins. These results define a system for investigating the biochemistry and regulation of cdc2 kinase activation by cyclin A

Breaking the Disk/Halo Degeneracy with Gravitational Lensing

The degeneracy between the disk and the dark matter contribution to galaxy rotation curves remains an important uncertainty in our understanding of disk galaxies. Here we discuss a new method for breaking this degeneracy using gravitational lensing by spiral galaxies, and apply this method to the spiral lens B1600+434 as an example. The combined image and lens photometry constraints allow models for B1600+434 with either a nearly singular dark matter halo, or a halo with a sizable core. A maximum disk model is ruled out with high confidence. Further information, such as the circular velocity of this galaxy, will help break the degeneracies. Future studies of spiral galaxy lenses will be able to determine the relative contribution of disk, bulge, and halo to the mass in the inner parts of galaxies.Comment: Replaced with minor revisions, a typo fixed, and reference added; 21 pages, 8 figures, ApJ accepte

NICMOS images of JVAS/CLASS gravitational lens systems

We present Hubble Space Telescope (HST) infrared images of four gravitational lens systems from the JVAS/CLASS gravitational lens survey and compare the new infrared HST pictures with previously published WFPC2 HST optical images and radio maps. Apart from the wealth of information that we get from the flux ratios and accurate positions and separations of the components of the lens systems that we can use as inputs for better constraints on the lens models we are able to discriminate between reddening and optical/radio microlensing as the possible cause of differences observed in the flux ratios of the components across the three wavelength bands. Substantial reddening has been known to be present in the lens system B1600+434 and has been further confirmed by the present infrared data. In the two systems B0712+472 and B1030+074 microlensing has been pinpointed down as the main cause of the flux ratio discrepancy both in the optical/infrared and in the radio, the radio possibly caused by the substructure revealed in the lensing galaxies. In B0218+357 however the results are still not conclusive. If we are actually seeing the two "true" components of the lens system then the flux ratio differences are attributed to a combination of microlensing and reddening or alternatively due to some variability in at least one of the images. Otherwise the second "true" component of B0218+357 maybe completely absorbed by a molecular cloud and the anomalous flux density ratios and large difference in separation between the optical/infrared and radio that we see can be explained by emission from either a foreground object or from part of the lensing galaxy.Comment: 10 pages, 4 figures (original higher resolution figures can be obtained at the e-mail above), to appear in MNRAS (accepted

The power spectrum from the angular distribution of galaxies in the CFHTLS-Wide fields at redshift ~0.7

We measure the real-space galaxy power spectrum on large scales at redshifts 0.5 to 1.2 using optical colour-selected samples from the CFHT Legacy Survey. With the redshift distributions measured with a preliminary ~14000 spectroscopic redshifts from the VIMOS Public Extragalactic Redshift Survey (VIPERS), we deproject the angular distribution and directly estimate the three-dimensional power spectrum. We use a maximum likelihood estimator that is optimal for a Gaussian random field giving well-defined window functions and error estimates. This measurement presents an initial look at the large-scale structure field probed by the VIPERS survey. We measure the galaxy bias of the VIPERS-like sample to be b_g=1.38 +- 0.05 (sigma_8=0.8) on scales k<0.2h/mpc averaged over 0.5<z<1.2. We further investigate three photometric redshift slices, and marginalising over the bias factors while keeping other LCDM parameters fixed, we find the matter density Omega_m=0.30+-0.06.Comment: Minor changes to match journal versio