scholarly journals The Drosophila kinesin-like protein KLP3A is a midbody component required for central spindle assembly and initiation of cytokinesis.

1995 ◽  
Vol 129 (3) ◽  
pp. 709-723 ◽  
Author(s):  
B C Williams ◽  
M F Riedy ◽  
E V Williams ◽  
M Gatti ◽  
M L Goldberg
Keyword(s):  
Motor Protein ◽  
Spindle Assembly ◽  
Male Meiosis ◽  
Cleavage Furrow ◽  
Critical Component ◽  
Central Spindle ◽  
Late Anaphase ◽  
Spindle Elongation ◽  
Anaphase B ◽  
Kinesin Superfamily

We describe here a new member of the kinesin superfamily in Drosophila, KLP3A (Kinesin-Like-Protein-at-3A). The KLP3A protein localizes to the equator of the central spindle during late anaphase and telophase of male meiosis. Mutations in the KLP3A gene disrupt the interdigitation of microtubules in spermatocyte central spindles. Despite this defect, anaphase B spindle elongation is not obviously aberrant. However, cytokinesis frequently fails after both meiotic divisions in mutant testes. Together, these findings strongly suggest that the KLP3A presumptive motor protein is a critical component in the establishment or stabilization of the central spindle. Furthermore, these results imply that the central spindle is the source of signals that initiate the cleavage furrow in higher cells.

Structure-function analysis of anaphase spindle elongation using isolated diatom spindles

1991 ◽  
Vol 49 ◽  
pp. 206-207
Author(s):  
W. Z. Cande ◽  
C.J. Hogan ◽  
M. Lee
Keyword(s):  
Morphological Changes ◽  
Function Analysis ◽  
Light And Electron Microscopy ◽  
Near Neighbor ◽  
Model Systems ◽  
Central Spindle ◽  
Spindle Poles ◽  
Spindle Elongation ◽  
Anaphase B

Diatom spindles are important model systems for describing the morphological changes associated with anaphase chromosome movement because the fibrous systems responsible for anaphase A (chromosome-to-pole movement) and anaphase B (spindle elongation) are spatially separate and the central spindle is a paracrystalline array of microtubules. The diatom central spindle, which is responsible for anaphase B, is constructed of two sets of interdigiting microtubules that originate from plate-like spindle poles and display specific near-neighbor interactions in the zone of microtubule overlap. The microtubules of each half-spindle are of relatively unifrom length such that the plus ends are clustered together in narrow zones at each edge of the zone of microtubule overlap. This has allowed us to monitor changes in extent of microtubule overlap in the light microscope with polarization optics. We have isolated spindles from synchronized populations of several species of dividing diatom cells to study the mechanochemistry of anaphase spindle elongation in vitro and to analyze the rearrangement of spindle components by light and electron microscopy during reactivation.

A requirement for the Abnormal Spindle protein to organise microtubules of the central spindle for cytokinesis inDrosophila

2002 ◽  
Vol 115 (5) ◽  
pp. 913-922 ◽  
Author(s):  
Maria Giovanna Riparbelli ◽  
Giuliano Callaini ◽  
David M. Glover ◽  
Maria do Carmo Avides
Keyword(s):  
Spindle Pole Body ◽  
Spindle Pole ◽  
Male Meiosis ◽  
Wild Type ◽  
Female Meiosis ◽  
Central Spindle ◽  
Spindle Poles ◽  
Late Anaphase ◽  
Mutant Cells ◽  
Sperm Aster

Drosophila abnormal spindle (asp) mutants exhibit a mitotic metaphase checkpoint arrest with abnormal spindle poles, which reflects a requirement for Asp for the integrity of microtubule organising centres (MTOCs). In male meiosis, the absence of a strong spindle integrity checkpoint enables asp mutant cells to proceed through anaphase and telophase. However, the central spindle region is not correctly organised and cells frequently fail to complete cytokinesis. This contrasts with meiosis in wild-type males where at late anaphase a dense array of microtubules forms in the central spindle region that has Asp localised at its border. We speculate that Asp is associated with the minus ends of microtubules that have been released from the spindle poles to form the central spindle. A parallel situation arises in female meiosis where Asp not only associates with the minus ends of microtubules at the acentriolar poles but also with the central spindle pole body that forms between the two tandem spindles of meiosis II. Upon fertilisation, Asp is also recruited to the MTOC that nucleates the sperm aster. Asp is required for growth of the microtubules of the sperm aster,which in asp mutants remains diminutive and so prevents migration of the pronuclei.

scholarly journals Analysis of the distribution of spindle microtubules in the diatom Fragilaria.

1978 ◽  
Vol 79 (3) ◽  
pp. 737-763 ◽  
Author(s):  
D H Tippit ◽  
D Schulz ◽  
J D Pickett-Heaps
Keyword(s):  
Spatial Arrangement ◽  
Spindle Pole ◽  
Serial Sections ◽  
Late Prophase ◽  
Central Spindle ◽  
Sources Of Error ◽  
Late Anaphase ◽  
Spindle Microtubules ◽  
Spindle Elongation ◽  
Intercellular Variability

The spindle of the colonial diatom Fragilaria contains two distinct sets of spindle microtubules (MTs): (a) MTs comprising the central spindle, which is composed of two half-spindles interdigitated to form a region of "overlap"; (b) MTs which radiate laterally from the poles. The central spindles from 28 cells are reconstructed by tracking each MT of the central spindle through consecutive serial sections. Because the colonies of Fragilaria are flat ribbons of contiguous cells (clones), it is possible, by using single ribbons of cells, to compare reconstructed spindles at different mitotic stages with minimal intercellular variability. From these reconstructions we have determined: (a) the changes in distribution of MTs along the spindle during mitosis; (b) the change in the total number of MTs during mitosis; (c) the length of each MT (measured by the number of sections each traverses) at different mitotic stages; (d) the frequency of different classes of MTs (i.e., free, continuous, etc.); (e) the spatial arrangement of MTs from opposite poles in the overlap; (f) the approximate number of MTs, separate from the central spindle, which radiate from each spindle pole. From longitudinal sections of the central spindle, the lengths of the whole spindle, half-spindle, and overlap were measured from 80 cells at different mitotic stages. Numerous sources of error may create inaccuracies in these measurements; these problems are discussed. The central spindle at prophase consists predominantly of continuous MTs (pole to pole). Between late prophase and prometaphase, spindle length increases, and the spindle is transformed into two half-spindles (mainly polar MTs) interdigitated to form the overlap. At late anaphase-telophase, the overlap decreases concurrent with spindle elongation. Our interpretation is that the MTs of the central spindle slide past one another at both late prophase and late anaphase. These changes in MT distribution have the effect of elongating the spindle and are not involved in the poleward movement of the chromosomes. Some aspects of tracking spindle MTs, the interaction of MTs in the overlap, formation of the prophase spindle, and our interpretation of rearrangements of MTs, are discussed.

scholarly journals Spindle assembly and cytokinesis in the absence of chromosomes during Drosophila male meiosis

2003 ◽  
Vol 160 (7) ◽  
pp. 993-999 ◽  
Author(s):  
Elisabetta Bucciarelli ◽  
Maria Grazia Giansanti ◽  
Silvia Bonaccorsi ◽  
Maurizio Gatti
Keyword(s):  
Chromosome Segregation ◽  
Spindle Assembly ◽  
Male Meiosis ◽  
Aurora B ◽  
Aurora B Kinase ◽  
Spindle Formation ◽  
Central Spindle ◽  
Morphological Transformations ◽  
Bipolar Spindles ◽  
Drosophila Mutants

Alarge body of work indicates that chromosomes play a key role in the assembly of both acentrosomal and centrosome-containing spindles. In animal systems, the absence of chromosomes either prevents spindle formation or allows the assembly of a metaphase-like spindle that fails to evolve into an ana-telophase spindle. Here, we show that Drosophila secondary spermatocytes can assemble morphologically normal spindles in the absence of chromosomes. The Drosophila mutants fusolo and solofuso are severely defective in chromosome segregation and produce secondary spermatocytes that are devoid of chromosomes. The centrosomes of these anucleated cells form robust asters that give rise to bipolar spindles that undergo the same ana-telophase morphological transformations that characterize normal spindles. The cells containing chromosome-free spindles are also able to assemble regular cytokinetic structures and cleave normally. In addition, chromosome-free spindles normally accumulate the Aurora B kinase at their midzones. This suggests that the association of Aurora B with chromosomes is not a prerequisite for its accumulation at the central spindle, or for its function during cytokinesis.

Interzonal microtubules are dynamic during spindle elongation

1990 ◽  
Vol 97 (2) ◽  
pp. 273-281 ◽  
Author(s):  
E. Shelden ◽  
P. Wadsworth
Keyword(s):  
Microtubule Assembly ◽  
Late Anaphase ◽  
Confocal Fluorescence ◽  
Fluorescence Light Microscopy ◽  
Fluorescence Light ◽  
Marked Decline ◽  
Microtubule Growth ◽  
Spindle Elongation ◽  
Anaphase B ◽  
Short Times

The pattern and extent of microtubule assembly during spindle elongation has been examined in PtK1 cells by microinjection of biotin-tubulin and immunolocalization of biotin-tubulin-containing microtubules using antibodies to biotin. PtK1 cells were microinjected at 30 degrees C, incubated for various intervals to allow incorporation of biotin-tubulin into microtubules, then lysed, fixed and stained for biotin-tubulin and total tubulin. When mid- to late anaphase cells were examined at short times post-injection, using conventional fluorescence light microscopy, rapid incorporation of biotin-tubulin was detected throughout the interzonal region, between the separating chromosomes, and in the spindle asters. Using confocal fluorescence microscopy, the segments of biotin-labeled microtubules in the interzonal region were found to be continuous with the distal, or plus-ends, of unlabeled microtubules. When teleophase cells were examined, a marked decline in the extent of incorporation was apparent. Quantitative analysis of the total length of labeled polymer in the interzonal region of cells from mid-anaphase through telophase further reveals that the extent of incorporation was maximal during late anaphase, and decreased during telophase. The measured rate of interzonal microtubule growth remained relatively constant during this period. Our results provide direct evidence for plus-end elongation of interzonal microtubules during spindle elongation and further reveal that interzonal microtubules are highly dynamic during late anaphase spindle elongation. The implications of these results for the mechanism of anaphase B are discussed.

scholarly journals Ipl1/Aurora-dependent phosphorylation of Sli15/INCENP regulates CPC–spindle interaction to ensure proper microtubule dynamics

2011 ◽  
Vol 194 (1) ◽  
pp. 137-153 ◽  
Author(s):  
Yuko Nakajima ◽  
Anthony Cormier ◽  
Randall G. Tyers ◽  
Adrianne Pigula ◽  
Yutian Peng ◽  
...  
Keyword(s):  
Microtubule Dynamics ◽  
Central Spindle ◽  
Microtubule Polymerization ◽  
Microtubule Stability ◽  
Late Anaphase ◽  
Chromosomal Passenger ◽  
Chromosome Separation ◽  
Spindle Elongation ◽  
Spindle Function ◽  
Spatiotemporal Control

Dynamic microtubules facilitate chromosome arrangement before anaphase, whereas during anaphase microtubule stability assists chromosome separation. Changes in microtubule dynamics at the metaphase–anaphase transition are regulated by Cdk1. Cdk1-mediated phosphorylation of Sli15/INCENP promotes preanaphase microtubule dynamics by preventing chromosomal passenger complex (CPC; Sli15/INCENP, Bir1/Survivin, Nbl1/Borealin, Ipl1/Aurora) association with spindles. However, whether Cdk1 has sole control over microtubule dynamics, and how CPC–microtubule association influences microtubule behavior, are unclear. Here, we show that Ipl1/Aurora-dependent phosphorylation of Sli15/INCENP modulates microtubule dynamics by preventing CPC binding to the preanaphase spindle and to the central spindle until late anaphase, facilitating spatiotemporal control of microtubule dynamics required for proper metaphase centromere positioning and anaphase spindle elongation. Decreased Ipl1-dependent Sli15 phosphorylation drives direct CPC binding to microtubules, revealing how the CPC influences microtubule dynamics. We propose that Cdk1 and Ipl1/Aurora cooperatively modulate microtubule dynamics and that Ipl1/Aurora-dependent phosphorylation of Sli15 controls spindle function by excluding the CPC from spindle regions engaged in microtubule polymerization.

scholarly journals Endosome mediated delivery of ceramide phosphoethanolamine (CPE) with unique acyl chain anchors to the cleavage furrow is essential for male meiosis cytokinesis

2021 ◽  
Author(s):  
Govind Kunduri ◽  
Si-Hung Le ◽  
Nagampalli Vijayakrishna ◽  
Daniel Blankenberg ◽  
Izumi Yoshihiro ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Acyl Chain ◽  
Structural Analog ◽  
Male Meiosis ◽  
Cleavage Furrow ◽  
Biophysical Properties ◽  
Contractile Ring ◽  
Central Spindle ◽  
Daughter Cells ◽  
Living Organisms

AbstractDivision of one cell into two daughter cells is fundamental in all living organisms. Cytokinesis, the final step of cell division, begins with the formation of an actomyosin contractile ring, positioned midway between the segregated chromosomes. Constriction of the ring with concomitant membrane deposition in a spatiotemporal precision generates a cleavage furrow that physically divides the cytoplasm. Unique lipids with specific biophysical properties have been shown to localize to midbodies however, their delivery mechanisms and biological roles were largely unknown. In this study, we show that Ceramide phosphoethanolamine (CPE), the structural analog of sphingomyelin, has unique acyl chain anchors in spermatocytes and is essential for meiosis cytokinesis. We found that disengagement of the central spindle from the contractile ring but not localization of phosphatidyl inositols (PIPs) at the plasma membrane was responsible for the male meiosis cytokinesis defect in CPE deficient animals. Further, we demonstrate that enrichment of CPE in Rab7 and Rab11 positive endosomes which in turn translocate to the cleavage furrows to promote cytokinesis. Our results implicate endosomal delivery of CPE to ingressing membranes is crucial for meiotic cytokinesis.

Sign in / Sign up

Export Citation Format

Share Document