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C de la Torre

Publications and source records attributed to C de la Torre.

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Proliferative index estimated by chromatin pattern analysis.

Chromatin pattern analysis at different density thresholds in Feulgen-stained meristems stimulated to proliferate by water inbibition, allows to estimate a proliferative potential index (PPI) which is a much earlier indicator of changes in proliferation than conventional labelling and mitotic indices. The PPI is but the ratio G1 cells to total 2C cells (or G2 to 4C cells, when cells also digress from the post-replicative stage of the cycle). The method is based in the fact that G1 and G2 cells have larger projected nuclear area and smaller dense chromatin area than their conterpart non-proliferating G0 and G0,2 cells.

Cell Cycle

Go and Go,2 cells as identified by their chromatin pattern in dormant and proliferating meristems.

Microdensitometry at different density thresholds of Feulgen-stained nuclei shows that both 2C and 4C dormant cells of the unsprouted root of Allium cepa L. bulbs have a smaller nuclear volume and a larger amount of dense chromatin than their counterpart G1 and G2 cells of the proliferating root. The analysis seems to differentiate between cells in the proliferate compartment from those in the non-proliferative compartment (Go and Go,2 cells).

Cell Cycle

Dynamics of the nuclear envelope during cell cycle in plants.

Stereology of Allium cepa root meristem cells was done to evaluate changes in the nuclear envelope during cell cycle. A naturally synchronous population was labelled as binucleate by caffeine inhibition of cytokinesis. Growth of the nuclear envelope preferentially occurs from mid G2 to the next mid G1, most probably in relation to the reforming sister nuclei after mitosis. On the other hand, the number of nuclear pores doubles from mid G1 to mid G2, their growth rate being higher in the first half of interphase (from mid G1 to mid S). Hence, the new nuclear envelope probably lacks nuclear pores, which appear later.

Cell Cycle

Compartmentalizing the S period.

In order to increase the resolution of interphase analysis we have developed a method which is an alternative to cytofluometric techniques for tissues where cell flow is not applicable. The method combines the estimation of cell frequency in G1, S, G2 and mitosis after a 3H-thymidine pulse with the grouping of interphase cells according to their DNA content, as estimated by cytophotometry in Feulgen stained nuclei. By superimposing both sets of data we get three different artificial compartments within the S period. As a biological test of the resolution reached, the method readily confirmed that hydroxyurea, after one cycle time, accumulates cycling cells of Allium cepa L. root meristems in early S.

Cell Compartmentation

Regulation of cycle progression in plant cells.

Induction of polynucleate cells in onion root meristems by inhibition of two sequential cytokineses is used to study controls operating in cell cycle progression. Triggering of both replication and metaphase occurs synchronously in nuclei sharing a common cytoplasm, independent of their ploidy or intracellular position. The replication rate appears to be activated by the simultaneous intracytoplasm presence of other replicating nuclei. On the other hand, central position of a nucleus in the cell as well as increase in ploidy leads to slowing down of replication rate. The relative advance and lag of S period in the different nuclei in a common cytoplasm is partially counter-balanced by differential times of G2. Moreover prophase lengthening in the fast interphase nuclei points ot a negative control exerted by the lagging nuclei mediated by cytoplasm. Finally, it could be worth emphasizing similarities in the control mechanism operating in cycle progression both in animal and plant cells.

Caffeine

Action of bleomycin on proliferating plant cells.

Bleomycin (10-(6) M) has been tested in Allium cepa L. meristems which are formed by a proliferating cell population growing under steady state conditions. Chromosome breaks were apparently induced by the antibiotic in cells in G2 period since anaphases with chromatid breaks were formed at a time shorter than G2 + prophase duration. Stimulation of entrance of G2 cells into mitosis is suggested both by an increase in the frequency of early prophases and by the study of waves of prophases in a synchronous subpopulation labelled by caffeine. Progression of other mitotic phases was unaffected. Nucleologenesis rate was increased by the antibiotic in a fashion resembling protein synthesis inhibitors. Protein synthesis is inhibited by 10-(6) M bleomycin to the same extent as 4 X 10(-6) M anisomycin. Both facts suggest that bleomycin has a direct inhibitory effect on protein synthesis in meristems. Given the nucleologenesis sensitivity to nucleolar RNA inhibition it is suggested that the antibiotic activity on nucleolar transcription is mediated through DNA.

Bleomycin