scholarly journals polo, a mitotic mutant of Drosophila displaying abnormal spindle poles

1988 ◽  
Vol 89 (1) ◽  
pp. 25-38 ◽  
Author(s):  
C.E. Sunkel ◽  
D.M. Glover
Keyword(s):  
High Frequency ◽  
Sex Chromosome ◽  
Living Cells ◽  
Male Meiosis ◽  
Mitotic Spindles ◽  
Spindle Poles ◽  
Immunofluorescence Studies ◽  
Meiotic Spindles ◽  
Circular Arrangement ◽  
Chromosome Disjunction

Neuroblast cells in larvae homozygous for mutant alleles of the locus polo show a high frequency of metaphases in which the chromosomes have a circular arrangement, and anaphase figures in which chromosomes appear to be randomly oriented with respect to at least one of the spindle poles. These defects appear to lead to the production of polyploid cells. Sex chromosome disjunction is affected in male meiosis, primarily in the second division, and the meiotic spindles of living cells are abnormal. One allele is a larval lethal, whereas another is semi-lethal with about 7% of homozygotes surviving as adults. Embryos from homozygous polo females have aberrant mitotic spindles that are highly branched and have broad poles. Immunofluorescence studies with an antibody that recognizes an antigen associated with the centrosome indicate that the organization of this organelle is disrupted in the mutant embryos.

scholarly journals Central-spindle microtubules are strongly coupled to chromosomes during both anaphase A and anaphase B

2019 ◽  
Vol 30 (19) ◽  
pp. 2503-2514 ◽  
Author(s):  
Che-Hang Yu ◽  
Stefanie Redemann ◽  
Hai-Yin Wu ◽  
Robert Kiewisz ◽  
Tae Yeon Yoo ◽  
...  
Keyword(s):  
Chromosome Segregation ◽  
Tomographic Reconstruction ◽  
Mitotic Spindles ◽  
Strongly Coupled ◽  
Central Spindle ◽  
Spindle Poles ◽  
Meiotic Spindles ◽  
Spindle Microtubules ◽  
Anaphase B ◽  
Chromosome Motion

Spindle microtubules, whose dynamics vary over time and at different locations, cooperatively drive chromosome segregation. Measurements of microtubule dynamics and spindle ultrastructure can provide insight into the behaviors of microtubules, helping elucidate the mechanism of chromosome segregation. Much work has focused on the dynamics and organization of kinetochore microtubules, that is, on the region between chromosomes and poles. In comparison, microtubules in the central-spindle region, between segregating chromosomes, have been less thoroughly characterized. Here, we report measurements of the movement of central-spindle microtubules during chromosome segregation in human mitotic spindles and Caenorhabditis elegans mitotic and female meiotic spindles. We found that these central-spindle microtubules slide apart at the same speed as chromosomes, even as chromosomes move toward spindle poles. In these systems, damaging central-spindle microtubules by laser ablation caused an immediate and complete cessation of chromosome motion, suggesting a strong coupling between central-spindle microtubules and chromosomes. Electron tomographic reconstruction revealed that the analyzed anaphase spindles all contain microtubules with both ends between segregating chromosomes. Our results provide new dynamical, functional, and ultrastructural characterizations of central-spindle microtubules during chromosome segregation in diverse spindles and suggest that central-spindle microtubules and chromosomes are strongly coupled in anaphase.

scholarly journals Central spindle microtubules are strongly coupled to chromosomes during both anaphase A and anaphase B

2019 ◽  
Author(s):  
Che-Hang Yu ◽  
Stefanie Redemann ◽  
Hai-Yin Wu ◽  
Robert Kiewisz ◽  
Tae Yeon Yoo ◽  
...  
Keyword(s):  
Chromosome Segregation ◽  
Tomographic Reconstruction ◽  
Mitotic Spindles ◽  
Strongly Coupled ◽  
Central Spindle ◽  
Spindle Poles ◽  
Meiotic Spindles ◽  
Spindle Microtubules ◽  
Anaphase B ◽  
Chromosome Motion

AbstractSpindle microtubules, whose dynamics vary over time and at different locations, cooperatively drive chromosome segregation. Measurements of microtubule dynamics and spindle ultrastructure can provide insight into the behaviors of microtubules, helping elucidate the mechanism of chromosome segregation. Much work has focused on the dynamics and organization of kinetochore microtubules, i.e. on the region between chromosomes and poles. In comparison, microtubules in the central spindle region, between segregating chromosomes, have been less thoroughly characterized. Here, we report measurements of the movement of central spindle microtubules during chromosome segregation in human mitotic spindles, and Caenorhabditis elegans mitotic and female meiotic spindles. We found that these central spindle microtubules slide apart at the same speed as chromosomes, even as chromosomes move towards spindle poles. In these systems, damaging central spindle microtubules by laser ablation caused an immediate and complete cessation of chromosome motion, suggesting a strong coupling between central spindle microtubules and chromosomes. Electron tomographic reconstruction revealed that the analyzed anaphase spindles all contain microtubules with both ends between segregating chromosomes. Our results provide new dynamical, functional, and ultrastructural characterizations of central spindle microtubules during chromosome segregation in diverse spindles, and suggest that central spindle microtubules and chromosomes are strongly coupled in anaphase.

scholarly journals Myosin-10 and actin filaments are essential for mitotic spindle function

2008 ◽  
Vol 182 (1) ◽  
pp. 77-88 ◽  
Author(s):  
Sarah Woolner ◽  
Lori L. O'Brien ◽  
Christiane Wiese ◽  
William M. Bement
Keyword(s):  
Mitotic Spindle ◽  
Actin Filaments ◽  
Spindle Pole ◽  
Mitotic Spindles ◽  
Unconventional Myosin ◽  
Spindle Poles ◽  
Chromosome Partitioning ◽  
Meiotic Spindles ◽  
Spindle Function

Mitotic spindles are microtubule-based structures responsible for chromosome partitioning during cell division. Although the roles of microtubules and microtubule-based motors in mitotic spindles are well established, whether or not actin filaments (F-actin) and F-actin–based motors (myosins) are required components of mitotic spindles has long been controversial. Based on the demonstration that myosin-10 (Myo10) is important for assembly of meiotic spindles, we assessed the role of this unconventional myosin, as well as F-actin, in mitotic spindles. We find that Myo10 localizes to mitotic spindle poles and is essential for proper spindle anchoring, normal spindle length, spindle pole integrity, and progression through metaphase. Furthermore, we show that F-actin localizes to mitotic spindles in dynamic cables that surround the spindle and extend between the spindle and the cortex. Remarkably, although proper anchoring depends on both F-actin and Myo10, the requirement for Myo10 in spindle pole integrity is F-actin independent, whereas F-actin and Myo10 actually play antagonistic roles in maintenance of spindle length.

scholarly journals Microtubule Flux and Sliding in Mitotic Spindles ofDrosophila Embryos

2002 ◽  
Vol 13 (11) ◽  
pp. 3967-3975 ◽  
Author(s):  
Ingrid Brust-Mascher ◽  
Jonathan M. Scholey
Keyword(s):  
Mitotic Spindles ◽  
Pole Motion ◽  
Spindle Poles ◽  
Balance Of Forces ◽  
Anaphase B

We proposed that spindle morphogenesis in Drosophilaembryos involves progression through four transient isometric structures in which a constant spacing of the spindle poles is maintained by a balance of forces generated by multiple microtubule (MT) motors and that tipping this balance drives pole-pole separation. Here we used fluorescent speckle microscopy to evaluate the influence of MT dynamics on the isometric state that persists through metaphase and anaphase A and on pole-pole separation in anaphase B. During metaphase and anaphase A, fluorescent punctae on kinetochore and interpolar MTs flux toward the poles at 0.03 μm/s, too slow to drive chromatid-to-pole motion at 0.11 μm/s, and during anaphase B, fluorescent punctae on interpolar MTs move away from the spindle equator at the same rate as the poles, consistent with MT-MT sliding. Loss of Ncd, a candidate flux motor or brake, did not affect flux in the metaphase/anaphase A isometric state or MT sliding in anaphase B but decreased the duration of the isometric state. Our results suggest that, throughout this isometric state, an outward force exerted on the spindle poles by MT sliding motors is balanced by flux, and that suppression of flux could tip the balance of forces at the onset of anaphase B, allowing MT sliding and polymerization to push the poles apart.

scholarly journals Interaction between Poly(ADP-ribose) and NuMA Contributes to Mitotic Spindle Pole Assembly

2009 ◽  
Vol 20 (21) ◽  
pp. 4575-4585 ◽  
Author(s):  
Paul Chang ◽  
Margaret Coughlin ◽  
Timothy J. Mitchison
Keyword(s):  
Binding Proteins ◽  
Magnetic Beads ◽  
Spindle Pole ◽  
Covalent Modification ◽  
Mitotic Spindles ◽  
Spindle Poles ◽  
Tankyrase 1 ◽  
Mitotic Spindle Pole

Poly(ADP-ribose) (pADPr), made by PARP-5a/tankyrase-1, localizes to the poles of mitotic spindles and is required for bipolar spindle assembly, but its molecular function in the spindle is poorly understood. To investigate this, we localized pADPr at spindle poles by immuno-EM. We then developed a concentrated mitotic lysate system from HeLa cells to probe spindle pole assembly in vitro. Microtubule asters assembled in response to centrosomes and Ran-GTP in this system. Magnetic beads coated with pADPr, extended from PARP-5a, also triggered aster assembly, suggesting a functional role of the pADPr in spindle pole assembly. We found that PARP-5a is much more active in mitosis than interphase. We used mitotic PARP-5a, self-modified with pADPr chains, to capture mitosis-specific pADPr-binding proteins. Candidate binding proteins included the spindle pole protein NuMA previously shown to bind to PARP-5a directly. The rod domain of NuMA, expressed in bacteria, bound directly to pADPr. We propose that pADPr provides a dynamic cross-linking function at spindle poles by extending from covalent modification sites on PARP-5a and NuMA and binding noncovalently to NuMA and that this function helps promote assembly of exactly two poles.

scholarly journals Spindle-to-cortex communication in cleaving, polyspermic Xenopus eggs

2015 ◽  
Vol 26 (20) ◽  
pp. 3628-3640 ◽  
Author(s):  
Christine M. Field ◽  
Aaron C. Groen ◽  
Phuong A. Nguyen ◽  
Timothy J. Mitchison
Keyword(s):  
Positive Feedback ◽  
Natural Experiment ◽  
Cell Cortex ◽  
Lateral Growth ◽  
Mitotic Spindles ◽  
Microtubule Stabilization ◽  
Spindle Poles ◽  
Early Embryos ◽  
Position And Orientation ◽  
Chromosome Passenger Complex

Mitotic spindles specify cleavage planes in early embryos by communicating their position and orientation to the cell cortex using microtubule asters that grow out from the spindle poles during anaphase. Chromatin also plays a poorly understood role. Polyspermic fertilization provides a natural experiment in which aster pairs from the same spindle (sister asters) have chromatin between them, whereas asters pairs from different spindles (nonsisters) do not. In frogs, only sister aster pairs induce furrows. We found that only sister asters recruited two conserved furrow-inducing signaling complexes, chromosome passenger complex (CPC) and Centralspindlin, to a plane between them. This explains why only sister pairs induce furrows. We then investigated factors that influenced CPC recruitment to microtubule bundles in intact eggs and a cytokinesis extract system. We found that microtubule stabilization, optimal starting distance between asters, and proximity to chromatin all favored CPC recruitment. We propose a model in which proximity to chromatin biases initial CPC recruitment to microtubule bundles between asters from the same spindle. Next a positive feedback between CPC recruitment and microtubule stabilization promotes lateral growth of a plane of CPC-positive microtubule bundles out to the cortex to position the furrow.

Role of microtubules and microtubule organizing centers on meiotic chromosome elimination in Sciara ocellaris

1997 ◽  
Vol 110 (6) ◽  
pp. 721-730 ◽  
Author(s):  
M.R. Esteban ◽  
M.C. Campos ◽  
A.L. Perondini ◽  
C. Goday
Keyword(s):  
Meiotic Chromosome ◽  
Chromosome Elimination ◽  
Male Meiosis ◽  
Meiotic Spindle ◽  
Monopolar Spindle ◽  
Meiotic Spindles ◽  
Cytoplasmic Microtubules ◽  
Spindle Microtubules ◽  
Meiosis I

Spindle formation and chromosome elimination during male meiosis in Sciara ocellaris (Diptera, Sciaridae) has been studied by immunofluorescence techniques. During meiosis I a monopolar spindle is formed from a single polar complex (centrosome-like structure). This single centrosomal structure persists during meiosis II and is responsible for the non-disjunction of the maternal X chromatids. During meiosis I and II non-spindle microtubules are assembled in the cytoplasmic bud regions of the spermatocytes. The chromosomes undergoing elimination during both meiotic divisions are segregated to the bud region where they associate with bundles of microtubules. The presence and distribution of centrosomal antigens in S. ocellaris meiotic spindles and bud regions has been investigated using different antibodies. gamma-Tubulin and centrin are present in the bud as well as in the single polar complex of first meiotic spindle. The results suggest that spermatocyte bud regions contain microtubule-organizing centres (MTOCs) that nucleate cytoplasmic microtubules that are involved in capturing chromosomes in the bud regions. The distribution of actin and myosin in the spermatocytes during meiosis is also reported.

scholarly journals Microinjection of fluorescent tubulin into dividing sea urchin cells.

1983 ◽  
Vol 97 (4) ◽  
pp. 1249-1254 ◽  
Author(s):  
P Wadsworth ◽  
R D Sloboda
Keyword(s):  
Sea Urchin ◽  
Daughter Cell ◽  
Fluorescent Protein ◽  
Living Cells ◽  
Lytechinus Variegatus ◽  
Linear Polymers ◽  
Spindle Poles ◽  
Image Intensification ◽  
Fluorescent Polymer

To follow the dynamics of microtubule (MT) assembly and disassembly during mitosis in living cells, tubulin has been covalently modified with the fluorochrome 5-(4,6-dichlorotriazin-2-yl)aminofluorescein and microinjected into fertilized eggs of the sea urchin Lytechinus variegatus. The changing distribution of the fluorescent protein probe is visualized in a fluorescence microscope coupled to an image intensification video system. Cells that have been injected with fluorescent tubulin show fluorescent linear polymers that assemble very rapidly and radiate from the spindle poles, coincident with the position of the astral fibers. No fluorescent polymer is apparent in other areas of the cytoplasm. When fluorescent tubulin is injected near the completion of anaphase, little incorporation of fluorescent tubulin into polymer is apparent, suggesting that new polymerization does not occur past a critical point in anaphase. These results demonstrate that MT polymerization is very rapid in vivo and that the assembly is both temporally and spatially regulated within the injected cells. Furthermore, the microinjected tubulin is stable within the sea urchin cytoplasm for at least 1 h since it can be reutilized in successive daughter cell spindles. Control experiments indicate that the observed fluorescence is dependent on MT assembly. The fluorescence is greatly diminished upon treatment of the cells with cold or colchicine agents known to cause the depolymerization of assembled MT. In addition, cells injected with fluorescent bovine serum albumin or assembly-incompetent fluorescent tubulin do not exhibit fluorescence localized in the spindle but rather appear diffusely fluorescent throughout the cytoplasm.

scholarly journals HIGH SPEED PHOTOMICROGRAPHY OF LIVING CELLS SUBJECTED TO SUPERSONIC VIBRATIONS

1931 ◽  
Vol 15 (2) ◽  
pp. 147-153 ◽  
Author(s):  
E. Newton Harvey ◽  
Alfred L. Loomis
Keyword(s):  
High Frequency ◽  
High Speed ◽  
Living Cells ◽  
Air Bubbles ◽  
Sound Waves ◽  
Camera System ◽  
Fluid Movement ◽  
High Frequency Sound ◽  
Frequency Sound ◽  
New Type

A new type of camera system is described capable of taking 1200 pictures a second through a microscope objective. Photographs showing the destruction of Arbacia eggs by high frequency sound waves indicate that the disintegration occurs in less than 1/1200 second. Eggs drawn out into spindle or tadpole shapes suggest that rapid movements of the fluid tearing the eggs may be responsible for the disintegration. Although no cavitated air bubbles show in the photographs, other experiments make it likely that the rapid fluid movement is the result of submicroscopic cavitation.

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