Confocal analysis of chromosome behavior in wheat × maize zygotes

Genome ◽  
2004 ◽  
Vol 47 (1) ◽  
pp. 199-205 ◽  
Author(s):  
Keiichi Mochida ◽  
Hisashi Tsujimoto ◽  
Tetsuo Sasakuma
Keyword(s):  
Metaphase Plate ◽  
Chromosome Elimination ◽  
Chromosome Movement ◽  
Hybridization Method ◽  
Chromosome Behavior ◽  
Whole Mount ◽  
Hybrid Embryo ◽  
Spindle Microtubules ◽  
Spindle Structure

Herein, we profile the first embryonic mitosis in a hybrid of wheat and maize by using a whole-mount genomic in situ hybridization method and immunofluorescence staining with a tubulin-specific antibody. We have successfully captured the dynamics of each set of parental chromosomes in the first zygotic division of the hybrid embryo 24-28 h after crossing. During the first zygotic metaphase, although both sets of parental chromosomes congressed into the equatorial plate of the zygote, the maize chromosomes tended to lag in comparison with the wheat chromosomes. During anaphase, each parental chromosome separated into its sister chromosomes; however, some of the maize chromosomes lagged around the metaphase plate as segregants. The maize sister chromosomes that did move toward the pole showed delayed and asymmetric movement as compared with the wheat ones. Immunological staining of tubulin revealed a bipolar spindle structure in the first zygotic metaphase. The kinetochores of the maize chromosomes that lagged around the metaphase plate did not attach to the spindle microtubules. These results suggest that factors on the kinetochores of maize chromosomes that are required to control chromosome movement are deficient in the zygotic cell cycle.Key words: whole-mount, GISH, chromosome elimination, hybrid embryogenesis.

scholarly journals Formaldehyde-based whole-mount in situ hybridization method for planarians

2009 ◽  
Vol 238 (2) ◽  
pp. 443-450 ◽  
Author(s):  
Bret J. Pearson ◽  
George T. Eisenhoffer ◽  
Kyle A. Gurley ◽  
Jochen C. Rink ◽  
Diane E. Miller ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Hybridization Method ◽  
Whole Mount

scholarly journals Meiosis in Sciara coprophila: structure of the spindle and chromosome behavior during the first meiotic division.

1982 ◽  
Vol 93 (3) ◽  
pp. 655-669 ◽  
Author(s):  
D F Kubai
Keyword(s):  
Chromosome Segregation ◽  
Chromosome Movement ◽  
Serial Sections ◽  
Chromosome Behavior ◽  
Monopolar Spindle ◽  
Prophase Nucleus ◽  
Fungus Gnat ◽  
Spindle Microtubules ◽  
Conical Radiation ◽  
Meiosis I

Light microscope descriptions of meiosis I in males of the fungus gnat Sciara coprophila suggested the presence of a monopolar spindle in which maternal and limited chromosomes move poleward while paternal chromosomes "back away" from the pole. The ultrastructural analysis reported here, based upon serial sections of cells in different stages of meiosis I, shows that the spindle is indeed monopolar with a distinctive differentiation, the polar complex, at one pole. This complex is the focus of a conical radiation of spindle microtubules. Kinetochores of paternal chromosomes face the complex and microtubules associated with these kinetochores run toward the complex. No kinetochore microtubules were discovered on maternal or limited chromosomes. When the position of paternal, maternal, and limited chromosomes is compared at various stages, it is found that limited chromosomes always remain near the polar complex, paternal chromosomes remain far from it and only maternal chromosomes move closer to the pole. Apparently, chromosome segregation does not depend on paternal chromosomes "backing away" from the pole, and the required movement of maternal chromosomes take place in the absence of kinetochore microtubules. In the prophase nucleus, limited and maternal chromosomes are already spatially separate from paternal chromosomes before the spindle forms. Thus, the monopolar spindle functions only to increase the distance between already segregated sets of chromosomes. An extensive system of microtubule-associated membranes outlines the spindle; the possibility that maternal chromosome movement is somehow related to the presence of this membrane is discussed.

scholarly journals Dynamic bonds and polar ejection force distribution explain kinetochore oscillations in PtK1 cells

2013 ◽  
Vol 201 (4) ◽  
pp. 577-593 ◽  
Author(s):  
Gul Civelekoglu-Scholey ◽  
Bin He ◽  
Muyao Shen ◽  
Xiaohu Wan ◽  
Emanuele Roscioli ◽  
...  
Keyword(s):  
Metaphase Plate ◽  
Quantitative Model ◽  
Force Distribution ◽  
Mitotic Chromosomes ◽  
Chromosome Behavior ◽  
Sister Chromatids ◽  
Ejection Force ◽  
Sister Kinetochore ◽  
Spindle Microtubules ◽  
Key Features

Duplicated mitotic chromosomes aligned at the metaphase plate maintain dynamic attachments to spindle microtubules via their kinetochores, and multiple motor and nonmotor proteins cooperate to regulate their behavior. Depending on the system, sister chromatids may display either of two distinct behaviors, namely (1) the presence or (2) the absence of oscillations about the metaphase plate. Significantly, in PtK1 cells, in which chromosome behavior appears to be dependent on the position along the metaphase plate, both types of behavior are observed within the same spindle, but how and why these distinct behaviors are manifested is unclear. Here, we developed a new quantitative model to describe metaphase chromosome dynamics via kinetochore–microtubule interactions mediated by nonmotor viscoelastic linkages. Our model reproduces all the key features of metaphase sister kinetochore dynamics in PtK1 cells and suggests that differences in the distribution of polar ejection forces at the periphery and in the middle of PtK1 cell spindles underlie the observed dichotomy of chromosome behavior.

scholarly journals Quantitative mRNA Imaging Throughout the Entire Drosophila Brain

2016 ◽  
Author(s):  
Xi Long ◽  
Jennifer Colonell ◽  
Allan M Wong ◽  
Robert H Singer ◽  
Timothée Lionnet
Keyword(s):  
In Situ Hybridization ◽  
Bessel Beam ◽  
Structured Illumination ◽  
Hybridization Method ◽  
Brain Sample ◽  
Whole Mount ◽  
Drosophila Brain ◽  
Subcellular Resolution ◽  
Expression Quantification

AbstractWe describe a fluorescence in situ hybridization method that permits detection of the localization and abundance of single mRNAs (smFISH) in cleared whole-mount adult Drosophila brains. The approach is rapid, multiplexable and does not require molecular amplification; it allows facile mRNA expression quantification with subcellular resolution on a standard confocal microscope. Using a custom Bessel Beam-Structured Illumination microscope (BB-SIM), we further demonstrate single-mRNA detection across the entire brain sample.

scholarly journals NITROUS OXIDE: EFFECTS ON THE MITOTIC APPARATUS AND CHROMOSOME MOVEMENT IN HELA CELLS

1973 ◽  
Vol 58 (1) ◽  
pp. 96-106 ◽  
Author(s):  
B. R. Brinkley ◽  
Potu N. Rao
Keyword(s):  
Nitrous Oxide ◽  
Hela Cells ◽  
Metaphase Plate ◽  
Microtubule Assembly ◽  
Chromosome Movement ◽  
Spindle Microtubule ◽  
Mitotic Apparatus ◽  
Ultrastructural Studies ◽  
Multipolar Spindles ◽  
Spindle Microtubules

When HeLa cells were grown in the presence of nitrous oxide (N2O) under pressure (80 lb/in2) mitosis was inhibited and the chromosomes displayed a typical colchicine metaphase (c-metaphase) configuration when examined by light microscopy. When the cells were returned to a 37°C incubator, mitosis was resumed and the cells entered G1 synchronously. Ultrastructural studies of N2O-blocked cells revealed a bipolar spindle with centriole pairs at each pole. Both chromosomal and interpolar (pole-to-pole) microtubules were also present. Thus, N2O, unlike most c-mitotic agents, appeared to have little or no effect upon spindle microtubule assembly. However, the failure of chromo somes to become properly aligned onto the metaphase plate indicated an impairment in normal prometaphase movement. The alignment of spindle microtubules was frequently atypical with some chromosomal microtubules extending from kinetochores to the poles, while others extended out at acute angles from the spindle axis. These ultrastructural studies indicated that N2O blocked cells at a stage in mitosis more advanced than that produced by Colcemid or other c-mitotic agents. Like Colcemid, however, prolonged arrest in mitosis with N2O led to an increased incidence of multipolar spindles.

scholarly journals GTSE1 regulates spindle microtubule dynamics to control Aurora B kinase and Kif4A chromokinesin on chromosome arms

2017 ◽  
Vol 216 (10) ◽  
pp. 3117-3132 ◽  
Author(s):  
Aaron R. Tipton ◽  
Jonathan D. Wren ◽  
John R. Daum ◽  
Joseph C. Siefert ◽  
Gary J. Gorbsky
Keyword(s):  
Mitotic Spindle ◽  
Meta Analysis ◽  
Metaphase Plate ◽  
Spindle Pole ◽  
Aurora B ◽  
Chromosome Movement ◽  
Aurora B Kinase ◽  
Nuclear Envelope Breakdown ◽  
M Phase ◽  
Spindle Microtubules

In mitosis, the dynamic assembly and disassembly of microtubules are critical for normal chromosome movement and segregation. Microtubule turnover varies among different mitotic spindle microtubules, dictated by their spatial distribution within the spindle. How turnover among the various classes of spindle microtubules is differentially regulated and the resulting significance of differential turnover for chromosome movement remains a mystery. As a new tactic, we used global microarray meta-analysis (GAMMA), a bioinformatic method, to identify novel regulators of mitosis, and in this study, we describe G2- and S phase–expressed protein 1 (GTSE1). GTSE1 is expressed exclusively in late G2 and M phase. From nuclear envelope breakdown until anaphase onset, GTSE1 binds preferentially to the most stable mitotic spindle microtubules and promotes their turnover. Cells depleted of GTSE1 show defects in chromosome alignment at the metaphase plate and in spindle pole integrity. These defects are coupled with an increase in the proportion of stable mitotic spindle microtubules. A consequence of this reduced microtubule turnover is diminished recruitment and activity of Aurora B kinase on chromosome arms. This decrease in Aurora B results in diminished binding of the chromokinesin Kif4A to chromosome arms.

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