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Calyculin A, an enhancer of myosin, speeds up anaphase chromosome movement.

ABSTRACT: Actin and myosin inhibitors often blocked anaphase movements in insect spermatocytes in previous experiments. Here we treat cells with an enhancer of myosin, Calyculin A, which inhibits myosin-light-chain phosphatase from dephosphorylating myosin; myosin thus is hyperactivated. Calyculin A causes anaphase crane-fly spermatocyte chromosomes to accelerate poleward; after they reach the poles they often move back toward the equator. When added during metaphase, chromosomes at anaphase move faster than normal. Calyculin A causes prometaphase chromosomes to move rapidly up and back along the spindle axis, and to rotate. Immunofluorescence staining with an antibody against phosphorylated myosin regulatory light chain (p-squash) indicated increased phosphorylation of cleavage furrow myosin compared to control cells, indicating that calyculin A indeed increased myosin phosphorylation. To test whether the Calyculin A effects are due to myosin phosphatase or to type 2 phosphatases, we treated cells with okadaic acid, which inhibits protein phosphatase 2A at concentrations similar to Calyculin A but requires much higher concentrations to inhibit myosin phosphatase. Okadaic acid had no effect on chromosome movement. Backward movements did not require myosin or actin since they were not affected by 2,3-butanedione monoxime or LatruculinB. Calyculin A affects the distribution and organization of spindle microtubules, spindle actin, cortical actin and putative spindle matrix proteins skeletor and titin, as visualized using immunofluorescence. We discuss how accelerated and backwards movements might arise.

SUBMITTER: Fabian L

PROVIDER: S-EPMC1847834 | biostudies-other | 2007

REPOSITORIES: biostudies-other

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Calyculin A, an enhancer of myosin, speeds up anaphase chromosome movement.

Fabian Lacramioara L Troscianczuk Joanna J Forer Arthur A

Cell & chromosome 20070324

Actin and myosin inhibitors often blocked anaphase movements in insect spermatocytes in previous experiments. Here we treat cells with an enhancer of myosin, Calyculin A, which inhibits myosin-light-chain phosphatase from dephosphorylating myosin; myosin thus is hyperactivated. Calyculin A causes anaphase crane-fly spermatocyte chromosomes to accelerate poleward; after they reach the poles they often move back toward the equator. When added during metaphase, chromosomes at anaphase move faster t ...[more]

PMID: 17381845

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Project description:IntroductionThe aim of this study was to identify movement synergies during normal-walking that can differentiate healthy adults in terms of gait adaptability at various speeds. To this end, the association between movement synergies and lower-limb coordination variability or Deviation Phase (DP) was investigated. This study also investigated the moderating effect of movement synergies on the relationship between DP and the smoothness of arm-swing motion (NJI).MethodA principal component analysis of whole-body marker trajectories from normal-walking treadmill trials at 0.8m/s, 1.2m/s and 1.6m/s was undertaken. Both DP and NJI were derived from approx. 8 minutes of perturbed-walking treadmill trials. Principal movement components, PMk, were derived and the RMS of the 2nd-order differentiation of these PMk (PAkRMS) were included as independent variables representing the magnitude of neuromuscular control in each PMk. Each PAkRMS were input into maximal linear mixed-effects models against DP and (DP x NJI) respectively. A stepwise elimination of terms and comparison of models using Anova identified optimal models for both aims.ResultsThe principal movement related to the push-off mechanism of gait (PA4RMS) was identified as an optimal model and demonstrated a significant negative effect on DP however this effect may differ considerably across walking-speeds. An optimal model for describing the variance in (DP x NJI) included a fixed-effect of PA6RMS representing Right-Left side weight transfer was identified.InterpretationThe hypotheses that individuals who exhibited greater control on specific kinematic synergies would exhibit variations during perturbed walking was substantiated. Supporting evidence for the role of movement synergies during the double-support phase of gait in proactively correcting balance was presented as well as the potential for this approach in targeted rehabilitation. The potential influence of leg dominance on gait adaptability was also discussed. Future studies should investigate further the role of walking-speed and leg dominance on movement synergies and look to generalize these findings to patient populations.

Dataset Information

Calyculin A, an enhancer of myosin, speeds up anaphase chromosome movement.

Publications

Calyculin A, an enhancer of myosin, speeds up anaphase chromosome movement.

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