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Biomedical subjects

C L Woldringh

Publications and source records attributed to C L Woldringh.

At least 73 records · Page 4Linked to original sources
PubMed1980

Changes in cell diameter during the division cycle of Escherichia coli.

Extensive measurements of steady-state populations of several Escherichia coli strains have consistently indicated that cell diameter decreases with increasing cell length. This was observed both after electron microscopy of air-dried cells and after phase-contrast microscopy of living cells. The analysis was made by considering separately the unconstricted cells and three classes (slight, medium, and deep) of constricted cells in the population. During slow growth, cells with the average newborn length were up to 8% thicker than unconstricted cells twice as long. This decrease in diameter is less at higher growth rates. Despite the small changes and the large variation of the diameter in any particular length class, significant negative correlations between diameter and length were obtained. Cell diameter increases again at the end of the cell cycle as indicated by an increase of average diameter in the three consecutive classes of constriction.

Bacillus subtilis↗
PubMed1978

Size variations and correlation of different cell cycle events in slow-growing Escherichia coli.

Cell lengths have been determined at which cycle events occur in the slow-growing Escherichia coli B/r substrains A, K, and F26. The radioautographic and electron microscope analyses allowed determination of the variations in length at birth, initiation and termination of DNA replication, and initiation of the constriction process and of cell separation. In all three substrains the standard deviation increased between cell birth and initiation of DNA replication. From there on, the standard deviation remained relatively constant until cell separation. These observations are consistent with the presence of a deterministic phase during the cell cycle in which the cell sizes at initation of DNA replication and at cell division are correlated.

Cell Cycle↗
PubMed1978

Chromosome replication rate and cell shape in Escherichia coli: lack of coupling.

The dimensions of Rep- cells of Escherichia coli K-12 were measured and compared with those of their Rep+ isogenic cells (both Thy-), Rep- cells cultivated identically were longer (but not wider), even though both strains were wider when the rate of chromosome replication was slowed down by lowering the thymine concentration supplied. This eliminates the possibility that cell shape is determined by this rate. Simulating Thy+ phenotype by adding deoxyguanosine resulted in shorter Rep- cells when growth was faster. This excludes a simple relationship between cell elongation and growth rate, but is consistent with a linear proportionality between the rate of surface synthesis and growth. Thymine limitation of fast-growing Thy- E. coli K-12 cells is shown to result in loss of their uniform shape and production of bizarre morphologies, apparently due to imbalanced synthesis of wall components.

Chromosomes, Bacterial↗
PubMed1977

Morphological analysis of the division cycle of two Escherichia coli substrains during slow growth.

Morphological parameters of the cell division cycle have been examined in Escherichia coli B/r A and K. Whereas the shape factor (length of newborn cell/width) of the two strains was the same at rapid growth (doubling time, tau, less than 60 min), with decreasing growth rate the dimensions of the two strains did change so that B/r A cells became more rounded and B/r K cells became more elongated. The process of visible cell constriction (T period) lasted longer in B/r A than in B/r K during slow growth, reaching at tau = 200 min values of 40 and 17 min, respectively. The time between termination of chromosome replication and cell division (D period) was found to be longer in B/r A than in B/r K. As a result, in either strain completion of chromosome replication seemed always to occur before initiation of cell constriction. Nucleoplasmic separation did not coincide with termination as during rapid growth but occurred in both strains within the T period, about 10 min before cell division.

Cell Division↗
PubMed1976

Significance of folded chromosomes released from amino-acid-starved Escherichia coli cells.

1. Folded chromosomes from amino-acid-starved Escherichia coli DG 75 cells are to a large extent released as envelope-bound complexes which sediment more rapidly than envelope-bound complexes from exponentially growth cells. A minor fraction (about 3%) represents relatively slow sedimenting envelope-free nucleoids. 2. Morphological analysis of the sensitivity of amino-acid-starved cells to the action of lysozyme and/or detergents indicates that these cells are less susceptible to lysis than exponentially grown cells. This results in the production of fast sedimenting envelope-bound complexes from non-dividing cells. We infer that it is not the amount of DNA, as suggested by Ryder and Smith (1974), but the low degree of envelope fragmentation that causes the high sedimentation rate. 3. After prolonged periods of starvation about 3% of cells in the process of division persist in the population. The results indicate that these cells release their (terminated) chromosomes in the envelope-free form. At this stage it is impossible to conclude whether these chromosomes are released because of their detachment from the membrane in situ (cf. Worcel and Burgi, 1974) or because of an enhanced susceptibility of dividing cells to lysis.

Amino Acids↗
PubMed1976

Factors affecting the release of folded chromosomes from Escherichia coli.

1. The envelope-bound folded chromosome prepared by gentle lysis of Escherichia coli K 12 DG 75 cells in the presence of 1.0 M sodium chloride was found to be a non-specific complex of DNA strands entangled in vesicle-like envelope structures. This envelope remnant is still more or less rod-shaped. 2. Both the envelope-bound and the envelope-free folded chromosomes arise directly from intact cells. Even under extreme conditions of lysis, envelope-bound chromosomes cannot be converted into envelope-free nucleoids.

Cell Fractionation↗
PubMed1976

Morphological analysis of nuclear separation and cell division during the life cycle of Escherichia coli.

Quantitative electron microscope observations were performed on Escherichia coli B/r after balanced growth with doubling times (tau) of 32 and 60 min. The experimental approach allowed the timing of morphological events during the cell cycle by classifying serially sectioned cells according to length. Visible separation of the nucleoplasm was found to coincide with the time of termination of chromosome replication as predicted by the Cooper-Helmstetter model. The duration of the process of constrictive cell division (10 min) appeared to be independent of the growth rate for tau equals 60 min or less but to increase with increase doubling time in more slowly growing cells. Physiological division, i.e., compartmentalization prior to physical separation of the cells, was only observed to occur in the last minute of the cell cycle. The morphological results indicate that cell elongation continues during the division process in cells with tau equals 32 min, but fails to continue in cells with tau equals 60 min.

Cell Division↗
PubMed1976

Localization of ampicillin-sensitive sites in Escherichia coli by electron microscopy.

Growth of Escherichia coli B/r ATCC 12407 (doubling time, 65 to 70 min) in the presence of 500 mug of ampicillin per ml for 15 to 20 min induces a sphere alongside the cell. The position was determined with respect to the length axis of the cell by electron microscopy. Although spheres may be found anywhere, some prominent sites do occur. In the shortest cells, which have a length of about 1.5 mum, they are found at the presumed new cell pole. In slightly older cells (length, about 1.8 mum), the position of the sphere is not well defined. Later on spheres occur predominantly at the cell center. In dividing cells (average length, 2.5 mum) a sphere may also occur at about one-quarter of the cell length. The position of the spheres bears resemblance to sites where a pulse of 3H-labeled diaminopimelic acid is incorporated into the peptidoglycan, as has been found by others.

Ampicillin↗
PubMed1976

Organization of the nucleoplasm in Escherichia coli visualized by phase-contrast light microscopy, freeze fracturing, and thin sectioning.

The organization of the nucleoplasm in Escherichia coli was studied by comparing the results obtained by freeze fracturing and thin sectioning. In addition to exponentially growing cells, we used chloramphenicol-treated cells which show a well-defined nucleoplasm, in the phase-contrast light microscope and can therefore function as a control for treatments necessary for electron microscopy. Two factors were found to determine the visibility of the nucleoplasm in freeze fractures: first, the state of lateral aggregation of deoxyribonucleic and fibrils, which is enhanced by postfixation with OsO4 according to the Ryter-Kellenberger technique; second, the presence of ice crystals. When their formation is prevented by the use of high concentration of freeze-protecting agents, the nucleoplasm appears as a smooth region in cells that have been prefixed. In unfixed cells, however, the freeze-protecting agent causes disappearance of the nucleoplasm by rearrangement of structures within the cell. This observation makes it hard to determine whether the deoxyribonucleic acid in vivo dispersed, as found after glutaraldehyde prefixation, or compact, as after OsO4 prefixation.

Cell Nucleus↗
PubMed1972

Effects of treatment with sodium dodecyl sulfate on the ultrastructure of Escherichia coli.

An electron microscopy study has been made of the effects of dissolution of the plasma membrane of Escherichia coli with sodium dodecyl sulfate (SDS) on the organization of the nucleoplasm and the cytoplasm. The alterations observed in time course experiments were related to absorbance changes and to release of macromolecules from the cells. As the cells became plasmolyzed, under the conditions used, the first visible effect of SDS was a collapse of the plasmolysis spaces. This was accompanied by a displacement of the nuclear material which then appeared in broad contact with the redeployed plasma membrane. This initial displacement of nuclear material to the cell border may indicate an association between the nucleoplasm and the plasma membrane. Upon further dissolution of the plasma membrane, the nuclear material receded from the cell margin and contracted into an axial filament. Meanwhile, the cytoplasm dissociated into an amorphous, Pronase-sensitive component and an electron-opaque, granular one sensitive to ribonuclease. The latter represented one continuous area of ribosomal structures surrounding the nucleoplasm, an organization which did not occur when the cells were inhibited with rifamycin before SDS treatment. During prolonged SDS interaction, approximately 65% of the cellular protein, 25% of the ribonucleic acid and 40% of the deoxyribonucleic acid were released from the cells concomitant with the disappearance of the amorphous cytoplasmic part, expansion of the ribosomal aggregate, and rearrangement of the nuclear material at the cell periphery. The observations support the contention that all ribosomal structures bear a direct relationship with the nucleoplasm.

Agar↗