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C N David

Publications and source records attributed to C N David.

At least 37 records · Page 2Linked to original sources

Male and female stem cells and sex reversal in Hydra polyps.

Single interstitial stem cells of male polyps of Hydra magnipapillata give rise to clones that differentiate either male or female gametes. To test the sexual stability of these clones, stem cells were recloned. The results indicate that stem cells from female clones are stable in their sexual differentiation capacity; male stem cells, by comparison, switch sexual phenotype at the rate of 10(-2) per cell per generation. As a result, female polyps contain only female stem cells; male polyps contain a mixture of male and female stem cells. A model is presented in which the sexual phenotype of Hydra polyps is controlled by (i) the switching rate of male and female stem cells and (ii) the repression of female differentiation by male stem cells.

Journal Article↗

Gland cells in Hydra: cell cycle kinetics and development.

The proliferative capacity of gland cells in Hydra attenuata was investigated. The results indicate that both gland cell proliferation and interstitial cell differentiation to gland cells contribute to the maintenance of the whole population. On the basis of [3H]thymidine incorporation and nuclear DNA measurements, gland cells consist of at least three different populations. One population consists of rapidly proliferating cells with a cell cycle of about 72 h. These cells are distributed throughout the body column. In the lower gastric region there is a population of non-cycling cells in G2 while in the upper gastric region there is a population of non-cycling cells in G1. About half the G1 population becomes a new antigen, SEC 1, which is typical of mucus cells.

Animals↗

Growth regulation in Hydra: relationship between epithelial cell cycle length and growth rate.

The relationship between epithelial cell production and growth rate was investigated in Hydra attenuata under different feeding regimes. The increase of epithelial cell number was compared to the duration of the epithelial cell cycle using standard methods of cell cycle analysis. The results indicate that cell cycle changes accompanying changes in feeding regime are not sufficient to explain the altered growth rate. Under heavy feeding regimes, epithelial cell production equals tissue growth rate. At low feeding level or under starvation conditions the epithelial cell cycle lengthens and growth rate of epithelial cell population is slowed. However, the cell cycle changes are insufficient to account for the reduction in tissue growth and thus there is an effective overproduction of epithelial cells amounting to 10% per day. Evidence suggests that these excess cells are phagocytized by neighboring cells in the tissue. Thus phagocytosis is directly or indirectly involved in regulating the growth of hydra tissue.

Animals↗

Loss of differentiating nematocytes induced by regeneration and wound healing in Hydra.

Cell death was observed in the nematocyte differentiation pathway in Hydra during head and foot regeneration. This death occurs throughout the regenerating piece, is transient in nature and is selective for committed stenotele and desmoneme precursors. Proliferating nematoblasts are unaffected. Cell death appears to be caused by release of a toxic factor rather than loss of a hormone required for differentiation, since regenerating pieces released a factor that inactivated differentiating nematocytes, and injured animals that had intact head and foot tissue also lost differentiating nematocytes. The inactivated nematocytes are removed by phagocytosis by epitheliomuscular cells.

Animals↗

A revision of the Dictyostelium discoideum cell cycle.

We have investigated the Dictyostelium discoideum cell cycle using fluorometric determinations of cellular and nuclear DNA contents in exponentially growing cultures and in synchronized cultures. Almost all cells are in G2 during both growth and development. There is no G1 period, S phase is less than 0.5 h, and G2 has an average length of 6.5 h in axenically grown cells. Mitochondrial DNA, which constitutes about half of the total DNA, is replicated throughout the cell cycle. There is no difference in the nuclear DNA contents of axenically grown and bacterially grown cells. Thus the long cell cycle in axenically grown cells is due to a lengthening of the G2 phase.

Cell Cycle↗

Dependence of cell-type proportioning and sorting on cell cycle phase in Dictyostelium discoideum.

The relationship between the cell cycle phase of vegetative amoebae and prestalk and prespore differentiation in the slug stage were investigated in the slime mould Dictyostelium discoideum. Cells were synchronized by release from the stationary phase. Samples were taken at various times during the course of a synchronous cell doubling, fluorescently labelled and mixed with cells of random cell cycle phase from exponentially growing cultures. The fate of the fluorescently labelled cells was recorded at the slug stage. Cells early in the cycle exhibit strong prestalk sorting; cells taken later in the cycle exhibit strong prespore sorting. The period of prestalk sorting occurs immediately following mitosis and lasts about 1 h in a cell cycle of about 7 h duration. Accompanying the altered sorting behaviour is a marked changed in the prestalk-prespore proportions in slugs formed from synchronized populations of cells. Cells synchronized early in the cycle form slugs with 55% prespore cells; cells synchronized late in the cycle form slugs with 90% prespore. The results are discussed in terms of models for the formation of the prestalk-prespore pattern in slugs.

Cell Cycle↗

Commitment during nematocyte differentiation in Hydra.

Nematocytes in Hydra differentiate from interstitial stem cells. Desmonemes differentiate mainly in the distal half of the body column while stenoteles differentiate predominantly in the proximal half. This difference was used to determine the timing of nematocyte-type commitment in the differentiation pathway. Cells were transferred from distal or proximal regions to all positions in the body column to test when the proportion of stenotele and desmoneme differentiation changed to reflect the new environment. In the first experiment, the distal region of the body column was isolated and permitted to regenerate a whole Hydra. In the second experiment, dissociated cells from distal or proximal regions were transplanted into regenerating aggregates of Hydra tissue. Both experiments effectively transferred cells from distal or proximal positions to positions throughout the body column. By comparing the kinetics of stenotele and differentiation with the time required for distal or proximal cells to differentiate stenoteles and desmonemes in accord with their new environment, it was possible to conclude that stenotele and desmoneme commitment occurs during the terminal cell cycle prior to nematocyte differentiation and not at the stem cell. Additional experiments indicated that the number of rounds of cell division preceding differentiation is fixed at the time stem cells enter the nematocyte pathway.

Animals↗

Oxygen gradients cause pattern orientation in Dictyostelium cell clumps.

We have investigated the formation of the prestalk-prespore pattern in Dictyostelium discoideum. Pattern formation occurs in clumps of Dictyostelium cells embedded in agar under a 100% oxygen atmosphere. Agar embedding allows us to control spatially the environment surrounding the cell clumps. Our results suggest that the ambient oxygen concentration plays a role in controlling the size of the multicellular mass. Further, oxygen gradients established across clumps embedded in agar or held in holes in a plastic barrier cause orientation of the prestalk-prespore pattern such that the anterior prestalk region forms at the highest end of the gradient. The results also indicate that developing cells have the ability to migrate up a gradient of oxygen.

Agar↗

Stem cell growth and differentiation in Hydra attenuata. I. Regulation of the self-renewal probability in multiclone aggregates.

Interstitial stem cells in Hydra are rapidly proliferating multipotent stem cells which continuously give rise to precursors for nerve and nematocyte differentiation. Growth of the stem cell population is controlled by the cell cycle time of the stem cells and the self-renewal probability, Ps (the fraction of stem cells in each generation which divide to yield more stem cells). In normal Hydra the stem cell generation time is 24 h and Ps = 0.6; under these conditions the stem cell population doubles in 3.5 days. In the present experiments we have systematically investigated the dependence of Ps on stem cell density. We culture stem cells in a feeder layer system consisting of aggregates of nitrogen-mustard (NM)-inactivated Hydra cells. In this system stem cell density can be varied over a wide range by changing the number of clone-forming units (CFU) added to the aggregates. We have measured the growth rate of the stem cell population and the cell cycle of stem cells in NM aggregates after 4--7 days of culture. From these data we calculate the value of Ps. The results indicate that the growth rate decreases 4-fold as the number of CFU seeded per aggregate increases from 10 to 400. Under these same conditions the cell cycle remains constant. The values of Ps calculated from these results indicate the Ps decreases from 0.75 in aggregates seeded with 10--30 CFU to 0.55 in aggregates seeded with 200--400 CFU. These results support a model in which Ps is controlled by negative feedback from neighbouring stem cells. In addition, our experiments indicate that Ps decreases during the growth of stem cell clones. When only a few stem cells are seeded in aggregates, they give rise to isolated clones distributed throughout the aggregate. Ps decreases markedly within such clones as they grow in size presumably due to increasing stem cell content of the clones. Since Ps in such isolated clones declines with growth, we infer that the local stem cell concentration is what controls Ps and that the spatial range of the negative feedback signal is short compared to the dimensions of NM aggregates.

Animals↗