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H R Bode

Publications and source records attributed to H R Bode.

At least 55 records · Page 3Linked to original sources

Gland cells arise by differentiation from interstitial cells in Hydra attenuata.

The origin of the gland cells in asexually reproducing adult hydra is unclear. There is evidence suggesting that the gland cells are a self-renewing population as well as contrary evidence suggesting that they must arise from another cell type. We have reexamined the question and found the latter to be the case. Analysis of ectoderm/endoderm chimeras in which the ectoderm was labeled with [3H]thymidine indicates a precursor for gland cells in the ectoderm which migrates into the endoderm. Analysis of grafts between labeled lower halves and unlabeled upper halves of animals indicates the migratory precursor is either a large or a small interstitial cell. Measurement of the cell cycle times of the gland cells and the epithelial cells provided further support. The cell cycle time of the gland cells appears to be longer than that of the epithelial cells of the endoderm throughout the animal. This means that in the steady-state growth condition of hydra tissue, the gland cells cannot maintain their population size simply by cell division. These results and other data suggest the following dynamics for the gland cell population. Gland cells arise by differentiation from large interstitial cells, undergo a limited number of cell divisions, and then become postmitotic.

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Formation of pattern in regenerating tissue pieces of Hydra attenuata. IV. Three processes combine to determine the number of tentacles.

The tentacles in hydra have characteristics of both spacing patterns and number-regulating patterns in that their number under some circumstances changes with the size of the animal and under others does not. To determine which type of processes could yield these results, an extensive analysis was undertaken of the size parameters pertinent to tentacle formation. To do this pieces of tissue, varying in shape and spanning a 30-fold size range, were excised and allowed to regenerate into complete animals. Three separate mechanisms were found to combine which resulted in the final number of tentacles: (1) the spread of the two-part head pattern to produce a competent band of tissue of a given size where the tentacles could form; (2) initiation of tentacle-forming centres by a spacing mechanism and (3) growth of the tentacles to a size proportional to the size of the animal.

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Growth regulation of the interstitial cell population in hydra. III. Interstitial cell density does not control stem cell proliferation.

The interstitial cells of hydra contain a stem cell population which produces several classes of differentiated cell types. A model has been proposed which governs the growth rate of the interstitial cell population. This model, based on the density of interstitial cells in the tissue, makes specific predictions about the relationships among this density, the proportion of stem cells in the interstitial cell population, the growth rate of the interstitial cell population, and the amount of nematocyte differentiation. Hydroxyurea treatments were used to experimentally reduce interstitial cell numbers, and the validity of these expected correlations was tested. The results demonstrate that the predictions of the interstitial cell density model were not upheld. Furthermore, the findings suggest that the interstitial cells are a heterogeneous population, containing some cells which are no longer stem cells but which do retain a limited capacity for proliferation. In the following paper (S. Heimfeld and H.R. Bode, 1986, Dev. Biol. 115, 59-68) we have proposed an alternative mechanism to explain the observed correlations, which incorporates this heterogeneity into amplification divisions of interstitial cells already committed to differentiation.

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Growth regulation of the interstitial cell population in hydra. IV. Control of nerve cell and nematocyte differentiation by amplification of non-stem interstitial cells.

The precursors for several differentiated cell types in hydra, such as nerve cells and nematocytes, arise from the interstitial cell population. Previously, it has been suggested that the interstitial cells represent a homogeneous stem cell population, and that both the rate of growth and the amount of differentiation are regulated strictly at the level of stem cell self-renewal and commitment. However, recent evidence does not support this viewpoint. In this paper we have proposed that the interstitial cell population is complex, containing both clonable stem cells and other cells which have a reduced division capacity. In response to hydroxyurea treatment, there is an amplification in the number of divisions that the non-stem interstitial cells undergo before differentiating. This amplification model is consistent with the correlations found in the preceding report (S. Heimfeld and H.R. Bode, 1986, Dev. Biol. 115, 51-58) and fits well with previously published data. An additional experiment which tests two specific predictions of this new model is presented.

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Germ cells in Hydra oligactis males. II. Evidence for a subpopulation of interstitial stem cells whose differentiation is limited to sperm production.

Animals containing germline-restricted interstitial cells were obtained by treating males from a clone of Hydra oligactis with hydroxyurea (HU) to lower the interstitial population to 1 or 2 cells per animal. A 3-day HU treatment produced animals whose interstitial cells did not form somatic cells, but did produce sperm. The isolation of these cells in HU-treated animals has lead us to propose that the interstitial cell population may contain subpopulations which possess different growth dynamics and developmental potentials. Through asexual propagation, we have cloned several animals containing only sperm precursor interstitial cells and have examined the growth and differentiation behavior of these cells in offspring propagated over a 2-year period. Evidence has been obtained which demonstrates (1) the extensive self-renewal capacity of the sperm precursor interstitial cells, and (2) the restricted differentiation capacity of these interstitial stem cells. Factors which affect cells entering and traversing the spermatogenic pathway are also presented.

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Plasticity in the nervous system of adult hydra. I. The position-dependent expression of FMRFamide-like immunoreactivity.

The plasticity of nerve cells expressing the neuropeptide FMRFamide was examined in adult hydra. Using a whole-mount technique with indirect immunofluorescence, the spatial pattern of neurons showing FMRFamide-like immunoreactivity (FLI) was visualized. These neurons were located in the tentacles, hypostome, and peduncle, but not in the body column or basal disc. Since every neuron in the nerve net is continuously displaced toward an extremity and eventually sloughed, the constant pattern of FLI+ neurons could arise in one of two ways. When displaced into the appropriate region, FLI- neurons are converted to FLI+ neurons, or FLI+ neurons arise by differentiation from interstitial cells. To distinguish between these two possibilities, interstitial cells, the multipotent precursors of the nerve cells, were eliminated by treatment with hydroxyurea or nitrogen mustard. Following head, or foot and peduncle, removal from these animals, the missing structures regenerated. The spatial pattern of FLI+ neurons reappeared in the newly regenerated head or peduncle. This shows FLI- neurons in the body column were converted to FLI+ when their position was changed to the head or the peduncle. When the peduncle was grafted into the body column, it was converted to basal disc or body column tissue, and FLI disappeared. The appearance and loss of FLI was always position dependent. These results indicate that the neurons in the mature nerve net can change their neuropeptide phenotype in response to changes in their position.

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Patterning of the head in hydra as visualized by a monoclonal antibody. I. Budding and regeneration.

A monoclonal antibody, CP8, has been isolated which displays a position-specific binding pattern to epithelial cells of Hydra oligactis. Antibody binding is restricted to the head of adult animals. When a new head develops during the budding process, CP8 binding is present in the area which will form the head well before morphological signs of it. Similarly, following decapitation as a new head regenerates, CP8 label appears covering a domed area at the apical end of the regenerate before tentacles evaginate delineating the head. When bud development or regeneration is complete, CP8 label is restricted to the new head. Experiments indicate the appearance of CP8 label during the formation of a head correlates closely with the patterning events which result in the determination of the tissue to form a head. The usefulness of CP8 as a diagnostic tool for exploring the dynamics of head pattern formation in hydra is discussed.

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Nerve cells in hydra: monoclonal antibodies identify two lineages with distinct mechanisms for their incorporation into head tissue.

The relationship between populations of nerve cells defined by two monoclonal antibodies was investigated in Hydra oligactis. A population of sensory nerve cells localized in the head (hypostome and tentacles) is identified by the binding of antibody JD1. A second antibody, RC9, binds ganglion cells throughout the animal. When the nerve cell precursors, the interstitial cells, are depleted by treatment with hydroxyurea or nitrogen mustard, the JD1+ nerve cells are lost as epithelial tissue is sloughed at the extremities. In contrast, RC9+ nerve cells remain present in all regions of the animal following treatment with either drug. When such hydra are decapitated to initiate head regeneration, the new head tissue formed is again free of JD1+ sensory cells but does contain RC9+ ganglion cells. Our studies indicate that (1) nerve cells are passively displaced with the epithelial tissue in hydra, (2) JD1+ sensory cells do not arise by the conversion of body column nerve cells that are displaced into the head, whereas RC9+ head nerve cells can originate in the body column, (3) formation of new JD1+ sensory cells requires interstitial cell differentiation. We conclude from these results that the two populations defined by these antibodies are incorporated into the h ad via different developmental pathways and, therefore, constitute distinct nerve cell lineages.

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Growth regulation of the interstitial cell population in hydra. I. Evidence for global control by nerve cells in the head.

The interstitial cells of hydra form a multipotent stem cell system, producing terminally differentiated nerve cells and nematocytes during asexual growth. Under well-fed conditions the interstitial cell population doubles in size every 4 days. We have investigated the possible role of nerve cells in regulating this behavior. Nerve cells are normally found in highest concentrations in the head region of hydra, while interstitial cells are primarily located in the body column. Our experimental approach was to construct, by grafting, animals in which the density of nerve cells varied in (1) the head region, or (2) the body column. The growth of the interstitial cell population was then measured in these hydra. The results indicate that differences in head nerve cell density are closely correlated with how fast the interstitial cell population increases in size. Variations in the level of either nerve cells or interstitial cells in the body column showed no such correlation. These findings suggest the existence of a signaling mechanism in the head region. This signal, which is a function of the density of head nerve cells, emanates from the head tissue and exerts global control on the growth of the interstitial cell population in the body column.

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Spermatogenesis in Hydra oligactis. I. Morphological description and characterization using a monoclonal antibody specific for cells of the spermatogenic pathway.

A morphological description of cells participating in sperm formation in Hydra oligactis males using a maceration procedure is presented. These descriptions are corroborated by the use of a monoclonal antibody, AC2, that binds to both a subpopulation of interstitial cells that appears to participate exclusively in gamete formation, and to all the gamete-differentiation products, including sperm intermediate cells, spermatids, and sperm. Use of the antibody as an interstitial cell marker has allowed an analysis of the behavior of the gamete-precursor (AC2+) subpopulation of interstitial cells during the asexual state and the early stages of gamete formation, when no differentiating sperm intermediates are present. The results indicate there is a gamete-producing subpopulation of interstitial cells which is present in low numbers in asexual males and undergoes extensive growth following the onset of spermatogenesis to give rise to sperm intermediate cells, and, eventually, the sperm. No input from the AC2- interstitial cells is required to account for this growth or subsequent sperm production. We speculate that the AC2+ interstitial cells may represent a unique subpopulation which is developmentally restricted to sperm production.

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A subset of cells in the nerve net of Hydra oligactis defined by a monoclonal antibody: its arrangement and development.

A monoclonal antibody, termed JD1, was generated that bound to a subset of the nerve cells in the hypostome and tentacles of Hydra oligactis. Using a whole-mount technique the spatial pattern of the subset of nerve cells and their processes could be clearly visualized using indirect immunofluorescence. The subset largely corresponds to the epidermal sensory cells. Using the same technique the development of the pattern during head regeneration and budding was examined. The appearance of the nerve cells coincides with the formation of both the tentacles and hypostome. When head regeneration does not occur, JD1+ cells do not appear suggesting the differentiation of JD1+ cells is an integral event in head formation dependent on antecedent patterning processes.

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Formation of pattern in regenerating tissue pieces of Hydra attenuata. III. The shaping of the body column.

Excised pieces of hydra body tissue of varying size and shape regenerate into cylinders with a head and foot at opposite ends. The numbers of cells along the axial and circumferential dimensions were determined before, during, and after regeneration. The main process in shaping the excised tissue into a body column was found to be a rearrangement of the cells. When regenerates of different size were measured, the proportions of the body columns were found to vary, such that the smaller the animal the squatter the body column was. The presence of the head in regenerates was necessary for the formation or maintenance of the cylindrical shape, while the size of the head determined the proportions of the cylinder. The formation of a gradient of adhesivity induced by the developing head is suggested as the basis for the rearrangement of the cells into the cylindrical form.

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Interstitial cell migration in Hydra attenuata. I. Quantitative description of cell movements.

The interstitial cell system of hydra contains multipotent stem cells which can form at least two classes of differentiated cell types, nerves and nematocytes. The amount of nerve and nematocyte production varies in an axially dependent pattern along the body column. Some interstitial cells can migrate, which makes it conceivable that this observed pattern of differentiation is not the result of regionally specified stem cell commitment, but rather arises by the selective movement of predetermined cells to the correct site prior to expression. To assess this latter possibility quantitative information on the dynamics of interstitial cell migration was obtained. Epithelial hydra were grafted to normal animals in order to measure (1) the number of cells migrating per day, (2) the location of these cells within the host tissue, and (3) the axial directionality of this movement. Tissue properties such as axial position and the density of cells within the interstitial spaces of the host were also tested for their possible influence on migration. Results indicate that there is a considerable traffic of migrating interstitial cells and this movement has many of the characteristics necessary to generate the position-dependent pattern of nerve differentiation.

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Interstitial cell migration in Hydra attenuata. II. Selective migration of nerve cell precursors as the basis for position-dependent nerve cell differentiation.

In Hydra, the interstitial cells constitute a multipotent stem cell system, forming nerve cells and nematocytes. Differentiation of these product cells varies in an axially dependent manner along the body column, and either of two simple hypotheses can explain this phenomena: (1) position-dependent stem cell determination or (2) selective migration of committed precursor cells. This latter hypothesis predicts that the migrating interstitial cell population is enriched with cells which are restricted in their proliferation and differentiation potential. To test this idea several developmental properties of the migrating cells were compared with those of the total interstitial cell population. The results indicate that those interstitial cells that migrate are not a random sample of the whole population. They contain a lower proportion of stem cells, as shown both by a slower growth rate and by a reduced capacity for clone formation. The migrating population is specifically enriched with interstitial cells that are committed to nerve differentiation. Coupled with previous measurements on migration rates, these findings suggest that the observed position-dependent pattern of nerve differentiation results from the selective redistribution of committed nerve cell precursors.

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Formation of pattern in regenerating tissue pieces of Hydra attenuata. II. Degree of proportion regulation is less in the hypostome and tentacle zone than in the tentacles and basal disc.

The relative sizes of the various structures in Hydra attenuata were compared over a broad range of animal sizes to determine in detail the ability to regulate proportions during regeneration. The three components of the head, namely hypostome, tentacles, and tentacle zone from which the tentacles emerge, the body column, and the basal disc were all measured separately. Ectodermal cell number was used as the measure of size. The results showed that the basal disc proportioned exactly over a 40-fold size range, and the tentacle tissue proportioned exactly over a 20-fold size range. In contrast, the hypostome and tentacle zone proportioned allometrically . With decreasing size, the hypostome and tentacle zone became an increasing fraction of the animal at the expense of body tissue, and in the very smallest regenerates at the expense of tentacle tissue. In their current form, the reaction-diffusion models proposed for pattern regulation in hydra are not consistent with the data.

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