Biomedical subjects

Norbert Vischer

Publications and source records attributed to Norbert Vischer.

3 recordsLinked to original sources

PubMed2006

Changes of initiation mass and cell dimensions by the 'eclipse'.

The minimum time (E) required for a new pair of replication origins (oriCs) produced upon initiating a round of replication to be ready to initiate the next round after one cell mass doubling, the 'eclipse', is explained in terms of a minimal distance (l(min)) that the replication forks must move away from oriC before oriCs can 'fire' again. In conditions demanding a scheduled initiation event before the relative distance l(min)/L(0.5) (L being the total chromosome length) is reached, initiation is presumably delayed. Under such circumstances, cell mass at the next initiation would be greater than the usual, constant Mi (cell mass per copy number of oriC) prevailing in steady state of exponential growth. This model can be tested experimentally by extending the replication time C using thymine limitation at short doubling times tau in rich media to reach a relative eclipse E/C < l(min)/L(0.5). It is consistent with results obtained in experiments in which the number of replication 'positions'n (= C/tau) is increased beyond the natural maximum, causing the mean cell size to rise continuously, first by widening, then by lengthening, and finally by splitting its poles. The consequent branching is associated with casting off a small proportion of normal-sized cells and lysing DNA-less cells. Whether or how these phenomena are related to peptidoglycan composition and synthesis are moot questions.

Cell Cycle↗

PubMed2006

Contribution of microtubule growth polarity and flux to spindle assembly and functioning in plant cells.

The spindle occupies a central position in cell division as it builds up the chromosome-separating machine. Here we analysed the dynamics of spindle formation in acentrosomal plant cells by visualizing microtubules labelled with GFP-EB1, GFP-MAP4 and GFP-alpha-tubulin and chromosomes marked by the vital dye SYTO82. During prophase, few microtubules penetrate the nuclear area, followed by nuclear envelope disintegration. During prometaphase, microtubules invading the nuclear space develop a spindle axis from few bipolar microtubule bundles, which is followed by spindle assembly. Using a novel quantitative kymograph analysis based on Fourier transformation, we measured the microtubule growth trajectories of the entire dynamic metaphase spindle. Microtubules initiating from spindle poles either pass through the metaphase plate to form interpolar microtubule bundles or grow until they reach chromosomes. We also noticed a minor fraction of microtubules growing away from the chromosomes. Microtubules grow at 10 microm/minute both at the spindle equator and at the spindle poles. Photobleached marks created on metaphase and anaphase spindles revealed a poleward tubulin flux. During anaphase, the velocity of tubulin flux (2 microm/minute) equals the speed of chromatid-separation. With these findings we identified spatially coordinated microtubule growth dynamics and microtubule flux-based chromosome-separation as important facets of plant spindle operation.

Arabidopsis Proteins↗

PubMed2005

Single-particle tracking of oriC-GFP fluorescent spots during chromosome segregation in Escherichia coli.

DNA regions close to the origin of replication were visualized by the green fluorescent protein (GFP)-Lac repressor/lac operator system. The number of oriC-GFP fluorescent spots per cell and per nucleoid in batch-cultured cells corresponded to the theoretical DNA replication pattern. A similar pattern was observed in cells growing on microscope slides used for time-lapse experiments. The trajectories of 124 oriC-GFP spots were monitored by time-lapse microscopy of 31 cells at time intervals of 1, 2, and 3 min. Spot positions were determined along the short and long axis of cells. The lengthwise movement of spots was corrected for cell elongation. The step sizes of the spots showed a Gaussian distribution with a standard deviation of approximately 110 nm. Plots of the mean square displacement versus time indicated a free diffusion regime for spot movement along the long axis of the cell, with a diffusion coefficient of 4.3+/-2.6x10(-5) microm2/s. Spot movement along the short axis showed confinement in a region of the diameter of the nucleoid ( approximately 800 nm) with an effective diffusion coefficient of 2.9+/-1.7x10(-5) microm2/s. Confidence levels for the mean square displacement analysis were obtained from numerical simulations. We conclude from the analysis that within the experimental accuracy--the limits of which are indicated and discussed--there is no evidence that spot segregation requires any other mechanism than that of cell (length) growth.

Algorithms↗